Lung cancer remains the leading cause of cancer-related deaths worldwide and non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer. With a variety of biological functions, Prohibitin1 (PHB1) has been proved tumor-associated. But there are conflicting data regarding the involvement of PHB1 in tumorigenesis and few studies regarding the role of PHB1 in lung cancer. The studies reported herein used a combination of clinical observations and molecular methods to investigate the possible role of PHB1 in NSCLC tissues and cell lines. PHB1 expression was evaluated by RT-PCR, RT-qPCR, Western blotting and immunohistochemistry analysis. Flow cytometric analysis was used to determine the surface expression profiles of PHB1 in lung cell lines. The results showed that PHB1 expression were generally increased in lung cancer tissues when compared with matched noncancerous tissues and closely related with tumor differentiation and lymph node invasion. PHB1 expression levels was also increased in three lung cancer cell lines (SK-MES-1, NCI-H157 and NCI-H292) as compared with BEAS-2B cells. Moreover, there were various subcellular localization of PHB1 in different lung cancer cells and the presence of PHB1 on the surface of lung cancer cells was significantly reduced. In conclusion, PHB1 expression is increased in NSCLC and the up-regulation of PHB1 is associated with clinically aggressive phenotype. The different subcellular localization of PHB1 in NSCLC cells and the loss of the membrane-associated PHB1 probably related to the tumorigenesis and progression of NSCLC and suggests that PHB1 may play different roles in various types of NSCLC.
Prohibitin 1; up-regulation; subcellular localization; non-small cell lung cancer
Failure of the embryo to implant now constitutes the major limiting step in IVF treatment. Successful implantation requires a vital embryo and an effective molecular dialogue with a ‘receptive’ endometrium. However, what precisely constitutes a receptive human endometrium remains poorly defined. Several observations have indicated that ovarian stimulation for IVF may impair endometrial receptivity. The histological approach to monitor endometrial maturation requires an invasive biopsy that excludes its use during the luteal phase of cycles in which implantation is the end-point objective as in IVF. In recent years, several studies have been reported that the removal of endometrial secretions immediately prior to embryo transfer provides sufficient material for analysis of markers of receptivity without disrupting embryo implantation. Therefore, analysis of protein patterns in endometrial secretion fluid may offer a relatively non-invasive means of assessing endometrial receptivity during fertility treatment cycles. Several studies have shown that protein profile expression in endometrial secretions undergo cyclical changes, and demonstrated significant differences between the natural cycle and stimulated cycle. These findings suggest that endometrial secretion analysis provide a novel means of investigating the effect of ovarian stimulation on the intrauterine environment at the time of embryo transfer, which may help to develop less disruptive ovarian stimulation protocols for IVF in the future.
Ovarian stimulation; endometrial secretion; endometrial receptivity; cytokine; in vitro fertilization
Confocal immunofluorescence is a valuable technique for the detection of relevant molecules in the pathogenesis of arthritis in rat models; however, it requires efficient processing of tissues including bone decalcification. The decalcification process must ensure the complete removal of calcium and also a proper preservation of cellular structures and, specially, the antigenicity of the tissue to allow the immunodetection of the molecules of interest. In the present study, we evaluated the effect of four different decalcifying solutions: the Morse´s solution, 10% EDTA (pH 7.4), 7% HCl/2% EDTA and 5% Nitric acid, as well as four different treatments of the tissues (including microwave irradiation) in the processes of decalcification for large pieces of adult rat bones (hind paw, fore paw, knee and column). We assessed the time of decalcification, the easiness of slicing, the morphological preservation and finally, the antigenicity of two different bone proteins (Osteopontin (OPN) and Osteocalcin (OC)) measured by its immunofluorescence intensity under controlled confocal microscopy conditions. Our results showed that the specimen size and the presence of skin are critical factors for the rate of decalcification, and no significant benefit was found if microwave irradiation is applied to the tissue. The comprehensive statistical analysis showed that the optimal solution for the detection of OPN and OC by confocal immunofluorescence is the 5% Nitric Acid, and followed by 10% EDTA (pH 7.4), Ana Morse solution and 7% HCl/2% EDTA.
Bone decalcification; arthritis rat model; confocal immunostainig
In this study, we determined the genotype distribution of two single nucleotide polymorphisms (SNPs) in secreted frizzled related protein 1 (SFRP1), rs3242 and rs921142, in a Caucasian bladder cancer case-control study. Allelic variants of the SNPs were determined using restriction fragment length polymorphism (RFLP) analysis and partly verified by sequencing analysis. Overall, DNA from 188 consecutive and 215 early-onset bladder cancer patients (≤45 years) as well as from 332 controls was investigated. Potential microRNA binding sites were determined for rs3242, and microRNA expression was analysed in cell lines and tumour specimens. We observed a remarkable distribution difference in rs3242 between bladder cancer patients and healthy controls (p=0.05). Additionally, we found a significant difference in genotype distribution (p=0.032), resulting from the difference of early-onset patients and the control group (p=0.007). The risk allele T showed increased frequency in the early-onset patient group (p=0.002). Genotype-dependent differences of microRNA binding capacity were predicted in SFRP1 mRNA for two microRNAs. Hsa-miR-3646 showed strong expression in cell lines and tumour tissue, whereas hsa-miR-603 exhibited weak expression. The rs921142 SNP showed no significant association with bladder cancer risk. This is the first study to describe an association of the SFRP1 SNP rs3242 and bladder cancer risk as well as the influence of rs3242 on genotype-dependent microRNA capacity on SFRP1 mRNA. The onset of bladder seems to be associated with the increased occurrence of the T-allele in rs3242.
SFRP1; SNP; bladder cancer; microRNA; Wnt signalling pathway
Our previous studies have demonstrated that cyclosporin A (CsA) promotes the proliferation and migration of human trophoblasts via the mitgen-activated protein kinase-3/1 (MAPK3/1) pathway. In the present study, we further investigated the role of nuclear factor (NF)-κB in the CsA-induced trophoblast proliferating cell nuclear antigen (PCNA) expression and migration, and its relationship to MAPK3/1 signal. Flow cytometry was used to analyze the expression of PCNA in trophoblasts. The migration of human primary trophoblasts was determined by wound-healing assay and transwell migration assay. Western blot analysis was performed to evaluate the activation of NF-κB p65 and NF-κB inhibitory protein I-κB in human trophoblasts. We found that treatment with CsA promotes PCNA expression and migration of human trophoblast in a dose-associated manner. Blocking of the MAPK3/1 signal abrogated the enhanced PCNA expression and migration in trophoblasts by CsA. In addition, CsA increased the phosphorylation of NF-κB p65 and the inhibitor I-κB in human trophoblasts in a time-related manner. Pretreatment with MAPK3/1 inhibitor U0126 abrogated the phosphorylation of NF-κB p65 and I-κB. Accordingly, the CsA-induced enhancement of PCNA expression and migration in trophoblasts was also decreased. This CsA-induced enhancement in the expression and migration of trophoblasts was abolished by pretreatment with pyrrolidine dithiocarbamate, a specific NF-κB inhibitor. Thus, our results suggest that CsA promotes PCNA expression and migration of human trophoblasts via MAPK-mediated NF-κB activation.
Cyclosporine A; trophoblast; PCNA; migration; signal transduction pathway
Interleukin-22 (IL-22) is a member of the IL-10 cytokine family and plays critical roles in inflammation, immune surveillance, and tissue homeostasis. However, whether IL-22 regulates the growth of endometrial stromal cells (ESCs), and participates in the pathogenesis of endometriosis remain unclear. In this study, we found that the expression of IL-22 and it receptors (IL-22R1 and IL-10R2) in eutopic endometrium and ectopic lesion of women with endometriosis was higher than that from healthy control. Recombinant human IL-22 (rhIL-22) stimulated the proliferation of ESCs in a dosage-dependent manner. On the contrary, anti-human IL-22 neutralizing antibody inhibited the proliferation of ESCs in vitro. The stimulatory effect of IL-22 on the proliferation of ESCs could be reversed by inhibitor of STAT5, ERK1/2 or AKT signal pathway. However, blocking STAT3, JNK or P38 signal pathway had no these effects. By Enzyme-linked immunosorbent assay (ELISA) and flow cytometry assay, we demonstrated the rhIL-22 not only stimulate the secretion of CCL2 and IL-8, but also significantly up-regulate the expression of IL-8 receptor CXCR1 on ESCs. Meanwhile, STAT5, ERK1/2 and or AKT signal inhibitors could abrogate the increase of CCL2, IL-8 and CXCR1 levels induced by rhIL-22. However, rhIL-22 had not similar influence on CCL2 receptor CCR2. Our current results suggested that the higher level of IL-22 and it receptors in eutopic endometrium may stimulate the expression of CCL2, IL-8/CXCR1, and further promote the growth of ESCs possibly through activating STAT5, MAPK/ERK1/2 and or AKT signal pathways, which may be involved in the occurrence and development of endometriosis.
IL-22; endometrial stromal cells; proliferation; CCL2; IL-8; endometriosis
Since hyperglycemia aggravates acute pancreatitis and also activates the receptor for advanced glycation endproducts (RAGE) in other organs, we explored if RAGE is expressed in the pancreas and if its expression is regulated during acute pancreatitis and hyperglycemia. Acute pancreatitis was induced by cerulein in untreated and streptozotocin treated diabetic mice. Expression of RAGE was analyzed by Western blot and immunohistochemistry. To evaluate signal transduction the phosphorylation of ERK1/ERK2 was assessed by Western blot and the progression of acute pancreatitis was monitored by evaluation of lipase activity and the pancreas wet to dry weight ratio. RAGE is mainly expressed by acinar as well as interstitial cells in the pancreas. During acute pancreatitis infiltrating inflammatory cells also express RAGE. Using two distinct anti-RAGE antibodies six RAGE proteins with diverse molecular weight are detected in the pancreas, whereas just three distinct RAGE proteins are detected in the lung. Hyperglycemia, which aggravates acute pancreatitis, significantly reduces the production of two RAGE proteins in the inflamed pancreas.
Receptor for advanced glycation endproducts (RAGE) isoforms; soluble RAGE; pancreatitis; hyperglycemia; inflammation; ERK1/ERK2 phosphorylation
Nonmetastatic gene 23-H1 (NME1, also known as nm23-H1) is a wide-spectrum tumor metastasis suppressor gene that plays an important role in suppressing the proliferation, adhesion and invasion of endometrial stromal cells (ESCs). The present study is undertaken to explore the mechanism by which NME1 in ESCs from endometriosis modulates the angiogenesis and herein participates in the pathogenesis of endometriosis. The expression of NME1 in the primary ESCs from normal endometrium without endometriosis was higher than that from eutopic endometrium and ectopic lesion with endometriosis. Silencing NME1 stimulated the secretion of angiogenic factors interleukin-8 (IL-8) and vascular-endothelial growth factor (VEGF) of the eutopic ESCs from women with endometriosis, and these effects could be abrogated by MAPK/ERK1/2 or AKT inhibitor. In addition, the supernatant of NME1-silenced ESCs increased the expression of angiogenesis-relative molecules CD62E and CD105, and promoted angiogenesis of human umbilical vein endothelial cells (HUVECs). Anti-human IL-8 or VEGF neutralizing antibody reversed the effect on angiogenesis of HUVECs induced by NME1-silenced ESCs. Our current results suggest that the abnormal lower expression of NME1 in ESCs secrete more IL-8 and VEGF through activation of MAPK/ERK1/2 and AKT signal pathways, up-regulate the level of CD62E and CD105, and finally lead to numerous angiogenesis of vascular endothelial cells in the endometriotic milieu, which is beneficial to the origin and development of endometriosis.
NME1; ESCs; HUVECs; angiogenesis; endometriosis
This study aimed to investigate the functional restoration of radiation-damaged salivary gland with human amniotic epithelial cells (hAECs) transplantation by intra-glandular injection. hAECs were isolated from the amnion tissues. After primary culture, the phenotype of hAECs of the second passage was identified by flow cytometry (FCM) and immunocytochemical staining. Then, hAECs were intra-glandularly injected into the irradiated glands of mice. At different time points after transplantation, the glands were collected for hematoxylin-eosin (HE) staining and immunofluorescence staining, and the saliva flow rate was also determined. Results showed these cells were positive for CD29, CD73 and CK19 and negative for CD44, CD34, CD45 and CD71. The transplanted hAECs in the recipient glands could differentiate into acinar-like cells and resulted in morphological and functional restoration of salivary gland.
Functional regeneration; salivary gland; irradiated; amniotic epithelial cells
We studied the clinicopathological and imaging characteristics of primary central nervous system diffuse large B-cell lymphomas (PCNS-DLBCL). Imaging, pathologic histology, and immunohistochemical staining characteristics were analyzed, and the immunoglobulin heavy and light chain gene rearrangement of 25 PCNS-DLBCL cases was examined. MicroRNA was extracted from 10 cases each of PCNS-DLBCL, extracerebral germinal center DLBCL (GC-DLBCL), and extracerebral non-GC-DLBCL (NGC-DLBCL); we conducted chip hybridization and comparatively analyzed the difference among the three. PCNS-DLBCLs typically involved no less than two cerebral lobes (10/25); the frontal lobe was affected most often (6/25). Target-shaped structures were observed in all PCNS-DLBCLs due to the proliferation of centroblast-like large lymphocytes surrounding the vessels. There was strong and diffuse immunostaining for CD20 and CD79a, and negative immunostaining for CD3, CD5, CD23, and cyclin D1 for all PCNS-DLBCLs. The percentage of cells with nuclear positivity for anti-Ki67 antibody ranged 50-90% (mean, 80%). Three, 19, and 22 PCNS-DLBCLs were CD10-, Bcl-6-, and melanoma ubiquitous mutated 1-positive, respectively. Twenty-four PCNS-DLBCLs were B-cell monoclonal. MicroRNA hybridization showed that 788 PCNS-DLBCL microRNAs/segments increased to at least twice that of NGC-DLBCLs, and 401 PCNS-DLBCL microRNAs/segments declined to less than half of that of NGC-DLBCLs. Six hundred and eleven PCNS-DLBCL microRNAs/segments increased to at least twice that of GC-DLBCLs, and 229 PCNS-DLBCL microRNAs/segments declined to less than half of that in GC-DLBCLs. PCNS-DLBCL typically affected multiple sites, tended to occur in older men, arose from activated B cells, had high B-cell monoclonality; its microRNA expression differed from that of NGC-DLBCL and GC-DLBCL.
Central nervous system; diffuse large B-cell lymphoma; gene rearrangement; microRNA
SARI is associated with the risk for several cancers, and loss of SARI expression is frequently found in aggressive and metastatic cancer. Limited evidence shows that SARI is a tumor suppressor gene, but the role of SARI in non-small cell lung cancer (NSCLC) has not been previously reported. This study was to investigate the SARI expression profile in surgically resected lung cancer tissues of Chinese patients by immunohistochemistry and evaluate the relationship between SARI expression and prognosis of lung cancer patients. Furthermore, SARI gene was transfected into lung cancer cells (A549), and the growth curve and cell healing of lung cancer cells were determined, aiming to investigate the influence of SARI on the growth and migration of lung cancer cells in vitro. Results showed that 103 of 195 (52.82%) tissues were positive for SARI. When compared with normal tissues, SARI expression significantly reduced in 50.26% of NSCLC tissues. Patients with negative or reduced SARI expression were more likely to have advanced lung cancer and lymph node metastasis. In squamous carcinoma and adenocarcinoma patients, the SARI expression had no relation with the survival time; However in one-on-one analysis SARI expression in tumor cells and adjacent tissues, patients which tumor cells SARI express reduced than adjacent tissues, survival time was significantly shorter than those without reduction in SARI expression (Log Rank test, p = 0.001). After transfection by SARI gene, the proliferation and migration of A549 cells were obviously inhibited (p < 0.001). These results demonstrate that decreased SARI expression may predict a poor prognosis in NSCLC patients, and SARI may serve as a prognostic biomarker and potential therapeutic target for lung cancer.
SARI; non-small cell lung cancer; prognosis; surgically resected cancer
Objective: This study aimed to investigate the expression and significance of ATF-3 in laryngeal squamous cell carcinoma (LSCC). Methods: Expression of ATF-3 was examined using immunohistochemistry methods in samples from 83 cases of LSCC carcinoma. MTT assay was used to detect proliferation of Hep-2 cells after ATF-3 knocked down by siRNA lentivirus. A mouse model was used to investigate the inhibitive role of ATF-3 siRNA in LSCC xenografts. Realtime RCR was used to detect Cyclin D1 expression after ATF-3 downregulation in Hep-2 cells. Results: The expression of ATF-3 was positively detected in all the 83 cases of LSCC cancer tissues while Only 4 cases of adjacent non-neoplastic tissues were detected with positive ATF-3 expression. The ATF-3 expression was statistically related with T stage, neck nodal metastasis, clinical stage and prognosis of LSCC. Both cell proliferation in vitro and tumor growth in vivo were suppressed after ATF-3 knockdown. Furthermore, the expression of Cyclin D1 was decreased after ATF-3 downregulation in Hep-2 cells. Conclusion: ATF-3 is involved in the progress of LSCC, and may provide clinical information for evaluation of prognosis of LSCC. The oncologic role of ATF-3 may be correlated with Cyclin D1 regulation.
Laryngeal squamous cell carcinoma; activating transcription factor 3; Cyclin D1
Objective: To investigate the inhibitory effect of plasmid-based survivin-specific short hairpin RNA and GRIM-19 on the growth of Hep-2 laryngeal cancer cells. Methods: The plasmid expressing survivin-specific short hairpin RNA (shRNA) and GRIM-19 (p-siRNA survivin/GRIM-19) was prepared and transfected into Hep-2 cells with Lipofectamine 2000. The mRNA and protein expression of surviving and GRIM-19 were measured with RT-PCR and western blot assay, respectively. MTT assay was employed to detect the proliferation of Hep-2 cells, and flow cytometry and AO/EB assay were done to determine the apoptosis of Hep-2 cells. Results: In the p-siRNA survivin/GRIM-19, the mRNA and protein expression of survivin was markedly reduced by 54.4% and 42.2%, and the reduction in protein expression of surviving was more obvious than that in the p-siRNA survivin group (37%) (P<0.05). The protein expression of GRIM-19 was markedly enhanced when compared with the control group (P<0.01). MTT assay revealed the proliferation of Hep-2 cells undergoing transfection with p-siRNA survivin/GRIM-19 was markedly inhibited, and the inhibition rate was as high as 79%, which was higher than that in the psi-survivin group (45%) and p-GRIM-19 group (35%). AO/EB assay and flow cytometry indicated that the apoptotic cells in the p-siRNA survivin/GRIM-19 group were dramatically increased as compared to the psi-survivin group and p-GRIM-19 group. Conclusion: The p-siRNA survivin/GRIM-19 has marked decrease in survivin expression and dramatic increase in GRIM-19 expression. Moreover, silencing of survivin and over-expression of GRIM-19 can significantly inhibit the growth and induce the apoptosis of Hep-2 in vitro and in vivo.
Co-expression plasmid; gene silencing; survivin; laryngeal cancer
Cancer treatment-related bone loss has become growing problematic, especially in breast and prostate cancer treated with hormone/endocrine therapy, chemotherapy and radiotherapy. However, bone loss caused by targeted therapy in cancer patients is largely unknown yet. In present study, a kinase inhibitors screen was applied for MC3T3-E1, a murine osteoprogenitor cell line, and seven kinase inhibitors (GSK1838705A, PF-04691502, Dasatinib, Masitinib, GDC-0941, XL880 and Everolimus) were found to suppress the cell viability with dose- and time-dependent manner. The most interesting is that many kinase inhibitors (such as lapatinib, erlotinib and sunitinib) can promote MC3T3-E1 cell proliferation at 0.01 μM. 4 out of 7 inhibitors were selected to perform the functional study and found that they lead to cell cycle dysregulation, treatments of PF-04691502 (AKT inhibitor), Dasatinib (Src inhibitor) and Everolimus (mTOR inhibitor) lead to G1 arrest of MC3T3-E1 cells via downregulation of cyclin D1 and p-AKT, whereas XL880 (MET and VEGFR inhibitor) treatment results in increase of sub-G1 and G2/M phase by upregulation of p53 protein. Our work provides important indications for the comprehensive care of cancer patients treated with some targeted drugs.
Cancer treatment-related bone loss; kinases inhibitors screening; osteoprogenitor cells
Angiogenesis is essential for invasive tumor growth and metastasis. Bevacizumab has been widely used for the treatment of non-small cell lung cancer (NSCLC). Various studies clearly demonstrate the relevance of Id-1 and VEGF in angiogenesis. The aim of this study was to establish the role of Id-1 expression in tumor progression and angiogenesis in relation to VEGF in NSCLC. Seventy five patients underwent surgery for lung cancers. The expressions of Id-1 and VEGF in NSCLC samples were determined by immunohistochemistry. Expression of Id-1 and VEGF showed a close correlation in NSCLC (p < 0.001). In addition, Id-1 strong expression group showed high incidence of metastasis in multivariate analysis (p = 0.028). Id-1 strong expression group had short metastasis-free survival (p = 0.008) and short recurrence-free survival (p = 0.027). Strong Id-1 expression in NSCLC had a poor prognosis in association with VEGF expression. Id-1 may function in tumor growth and progression via angiogenesis. Therefore, Id-1 is considered to be a candidate for new therapeutic target and a prognostic factor in NSCLC.
Id proteins; vascular endothelial growth factor (VEGF); non-small cell lung cancer; prognosis; metastasis
A high frequency of mutations at the PTEN locus has been noticed in carcinoma of lung. However, the role of PTEN alternations and its association with outcome variables in the genesis of lung carcinoma are not understood fully. The purpose of our study was to examine the impact of EGFR, TGF-α, P-AKT and PTEN in the genesis of non-small cell lung cancer (NSCLC). Total numbers of 66 histopathologically confirmed cases of NSCLC and 10 cases of benign control samples embedded with wax were studied. We assessed EGFR, TGF-α and P-AKT by the use of specific antibody through immunohistochemistry as directed by the manufacturer, and detected PTEN expression by in situ hybridization. There were progressive loss of PTEN expression and significant increasing in EGFR, TGF-α, P-AKT expression from benign samples to NSCLC (p<0.05). The overexpression of EGFR, TGF-α, P-AKT and loss of PTEN expression were correlated to differentiation extent of cancer tissue, metastasis of lymph nodes and histological classification. Thus, alteration of EGFR, TGF-α, P-AKT and PTEN are likely important molecular events in pathogenesis and carcinogenesis of NSCLC.
Non-small cell lung cancer (NSCLC); P-AKT; TGF-α; EGFR; PTEN
Objective: Neoadjuvant chemotherapy (NACT) followed by cytoreduction has now become a part of standard care for patients with advanced ovarian cancer. Cytologic changes of the cancer cells induced by NACT, however, sometimes may cause confusion in terms of pathologic diagnosis and therefore inappropriate management. The objective of this study was to characterize the histologic or cytologic features of the ovarian cancers from those patients who received NACT in order to improve the diagnostic accuracy and reduce unnecessary clinical workup. Methods: Specimens from 120 patients with advanced ovarian cancer who received NACT were studied. All 120 cases had either cytologic samples from ascites (n=108) or fine needle aspiration (n=12) and the diagnosis of consistent with cancers of ovarian origin was made prior to NACT. There were 70 (58.3%) patients received subsequent tumor debulking surgery after NACT. The time frame between NACT and debulking surgery ranged from 28 to 65 with an average of 45 days. Among the 70 cases with cytoreductive surgery, 48 cases containing both pre-NACT cytology/histology and subsequent debulking specimens were suitable for the study. All 48 post-NACT ovarian cancers were reviewed and the characteristic pathologic features in gross were summarized. Microscopic evaluation and immunohistochemical stainings with antibodies against ER, PR, p53, WT1, PAX8, CK7, CK20, and CDX2 were performed to confirm the primary site and histologic type of the cancers. Results: Grossly, tumor size within the ovaries from those debulking specimens ranged from 2.3 to 6.5 cm in greatest dimension. The cancers were mainly solid (average of 65%) and cystic areas had more or less hemorrhagic appearance. Extensive tumor necrosis and some with fibrosis were present. Microscopically, the non-necrotic cancer cells were arranged in cords, islands and sometimes as scattered single large cells with large amount of eosinophilic cytoplasm with vacuoles. The viable cancer cells contained more or less vacuolated cytoplasm in almost all post chemotherapy cases. Multinucleated tumor giant cells were noted in close to half of the cases. The cancer cells commonly had large hyperchromatic bizarre nuclei with coarse chromatin clumping and sometimes prominent nucleoli. Due to the unusual cytologic changes after NACT, there was a concern of non-ovarian origin or the different histologic type of the cancers. Therefore, immunohistochemical (IHC) staining with the antibodies against ER, PR, PAX8, WT1, CK7, CK20, and CDX2 was performed in all 48 pairs of the cases. The 48 paired samples showed identical immunophenotype in pre- and post-NACT cancers, confirming there was no metastatic or new primary cancer involved in the study. Conclusions NACT can apparently induce significant cytologic/histologic changes in ovarian cancer. Aware of such NACT induced changes will be useful to make correct diagnosis for those patients who have received NACT. IHC with appropriate panels of the antibodies will be helpful to aid the diagnosis, particularly when nuclear change is dramatic and the clinical history of ovarian cancer is not available.
Ovarian cancer; cytological changes; chemotherapy; immunohistochemical
Objective: Now there are more and more evidences that Cyclooxygenase-2 (COX-2) plays an important role in angiogenesis of endometriosis (EMs). Vascular endothelial growth factor (VEGF) has a potent angiogenic activity. However, it is worth studying about the regulating mechanism of COX-2/COX-1 and VEGF in the development of human endometriosis in vitro. The current study was designed to investigate the effect of 4 cytokines on COX-2/COX-1 expression and the effect of IL-1β on VEGF release in human endometriosis stromal cells (ESC), and to explore the related signaling pathways involved in vitro. Methods: Isolation, culture and identification of ESC. Cells were treated with 4 cytokines, and the inhibitor mitogen-activated protein-Erk (MEK) and the inhibitor p38 mitogen-activated protein kinase (MAPK) prior to adding cytokine IL-1β. COX-2 protein expression was measured by western blot and VEGF secretion was determined by ELISA. Results: Among four kinds of cytokines, IL-1β treatment increased COX-2 protein expression and VEGF release in three ESC, and TNF-α had the same effect on COX-2 protein level as IL-1β only in ectopic and eutopic ESC, and MCSF had only slight effect on ectopic ESC. In contrast, cytokines had no effect on COX-1 expression. We also demonstrated that MAPK reduced the synthesis of COX-2 by IL-1β induced. COX-2 inhibitor reduced VEGF release by IL-1β induced. Conclusions: i) In human ESC in vitro, IL-1β up-regulated the COX-2 expression through the activation of p38 MAPK pathway, and not to COX-1. ii) Up-regulation of VEGF level by IL-1β treatment was found in human endometriosis stromal cell and COX-2 inhibitor was involved in this process.
Endometriosis stromal cells (ESC); cyclooxygenase-2 (COX-2); vascular endothelial growth factor (VEGF); cytokines; signal pathways; in vitro
Background: NIN/RPN Binding protein 1 homologue (NOBp1), encoded by NOB1 gene, was reported to play an essential role in the oncogenesis and prognosis of carcinomas. We conducted a study to reveal its expression and clinical significance in breast infiltrating ductal carcinoma. Methods: To explore the relationship between NOB1 expression and the clinical TNM (cTNM), 162 patients who undergone surgery were involved in the study. Compared to healthy tissues, abnormal localization and higher level of NOB1 in tumor cells was observed by Immunohistochemistry staining. Real-time PCR and western-blotting verified the up-regulation of NOB1 in carcinoma individuals. Results: A significant correlation between high level of NOB1 and the T stage, lymph node metastasis and cTNM was shown. Furthermore, patients with higher level of NOB1 predicted a declined overall survival (OS). Notably, multivariate analyses by Cox’s proportional hazard model revealed that expression of NOB1 was an independent prognostic factor in breast infiltrating ductal carcinoma. Conclusions: In summary, our present study clarify that the aberrant expression of NOB1 in breast infiltrating ductal carcinoma is possibly involved with tumorigenesis and development, and the NOB1 protein could act as a potential biomarker for prognosis assessment of breast infiltrating ductal carcinoma. Related mechanism is worthy of further investigation.
Breast cancer; NOB1 protein; immunohistochemistry; tissue microarray
Phyllodes tumors (PTs) are classified as fibroepithelial tumors and their histologic grade is determined primarily by the features of the stromal component. In this study, we examined the expression profiles of autophagy-related proteins in the stromal component of PTs and analyzed their clinical implications. We selected 204 human PT samples which were excised and diagnosed at Severance Hospital from 2000 to 2008 and created tissue microarray (TMA) blocks. Immunohistochemical assays for autophagy-related proteins (beclin-1, LC3A, LC3B, and p62) were then performed on these samples. The surgical specimens from higher grade PTs less frequently displayed cytoplasmic expression of beclin-1, LC3A, LC3B, and p62 in the stromal component (p<0.001). In univariate analysis, the following profiles were associated with shorter disease-free survival and overall survival: nuclear beclin-1 positivity in the stromal component (p=0.013 and p=0.044, respectively), LC3A positivity in the stromal component (p<0.001 and p<0.001, respectively), and p62 positivity in the stromal component (p=0.012 and p=0.004, respectively). In conclusion, we determined that increased activity of autophagy-related proteins correlated with a higher histologic grade and poorer prognosis in PTs. These results lead us to conclude that the autophagy activity of the stromal cells plays a key role in the progression of PTs.
Breast; phyllodes tumor; autophagy
Immune complex-mediated complement activation through the classic pathway plays a key role in the pathogenesis of lupus nephritis (LN). C4d deposition in renal tissue reflects the prognosis of systemic lupus erythematosus (SLE). The aim of the current study is to investigate the pathogenesis and clinicopathologic significance of glomerular C4d deposition in LN. We retrospectively analyzed clinical and histopathological data of 20 SLE patients with renal biopsy-proven LN and 10 non-SLE renal biopsy samples as control. LN biopsies showed varying degrees of glomerular C4d staining associated with immune complex deposits, IgG (p = 0.015), C1q (p = 0.032) and C3 (p = 0.049). 7 LN biopsies had all of C4d, C1q and C3 deposits in their glomeruli, indicative of the activation of the classical pathway, whereas 2 LN biopsies had C4d and C3 deposits without accompanying C1q deposits, indicating the activation of the lectin pathway. Glomerular C4d deposition was correlated with the LN subtype (p < 0.001). In particular, a diffusely intense and coarsely granular pattern of C4d deposition in all glomeruli was detected in class V membranous LN. However, glomerular C4d deposition was correlated with neither disease activity of SLE nor histological activity and chronicity of LN. In conclusion, the activation of the lectin pathway as well as the classical pathway seems to play a crucial role in the pathogenesis of LN. Glomerular C4d staining could be helpful for diagnosing class V membranous LN, although glomerular C4d deposition does not reflect SLE disease activity and histological activity and chronicity.
Lupus nephritis; C4d; classical pathway; lectin pathway; disease activity
In situ hybridization (ISH) was performed on paraffin-embedded tissues to detect multiple high-risk human papillomavirus (HPV) subtypes in 27 cases of cervical adenocarcinoma in situ (AIS) and adenocarcinoma (CA) specimens. These results were compared with those of HPV detection by HPV-PCR genotyping and p16 immunohistochemistry in the same specimens. Of the 27 cases, 17 (63%) showed HPV-DNA by HPV-ISH, including 3 metastatic lesions. HPV-DNA was detected in 18 cases (67%) by PCR. The concordance rate between HPV-ISH and HPV-PCR genotyping was 74% with moderate agreement (Kappa value, 0.41). HPV-16 was identified in 5 cases, HPV-18 in 2 cases, and HPV-45 in 1 case. Combining the results of HPV-ISH and HPV-PCR/genotyping, 22 cases (81.5%) were considered HPV positive. Immunohistochemical staining of p16 indicated that 25 (93%) cases were positive; however, 4 of these cases were HPV-negative by both PCR and ISH. Combining HPV-ISH and HPV-PCR/genotyping techniques demonstrated a high sensitivity of HPV detection in FFPE tissues from cervical glandular neoplasias. In contrast, p16 immunohistochemistry seemed to have a low specificity for determining HPV status in cervical glandular neoplasia. HPV-ISH is useful for recognizing the distribution of HPV in AIS and CA tissues and visualizing signal patterns, and may be a useful tool to confirm the cervical origin of neoplasias and metastatic lesions.
Uterine cervical cancer; adenocarcinoma; human papillomavirus; in situ hybridization; PCR; p16INK4A
Background: Hibernoma is a rare benign fat-forming soft tissue tumor that differentiates similar to brown fat, hence an origin from remnants of fetal brown adipose tissue has been proposed. Mainly young adults are affected, usually without significant clinical symptoms. Material and methods: We report on four patients with hibernomas, who were treated at our hospital during the last 10 years. The clinicopathologic and immunohistochemical features are presented and treatment and follow-up data discussed. Results: Patients were 2 women and 2 men aged 21-67 years (mean: 45 yrs) who presented with a slowly growing, painless mass. The anatomic location was the thigh, upper arm, lateral thoracic wall and paravertebral soft tissue. Two of them were diagnosed preoperatively through a percutaneous core needle biopsy and the other two underwent surgery because of high clinical and radiological suspicion of liposarcoma. The tumor’s size ranged from 7 cm to 15.5 cm (mean: 11 cm). All were deep-seated subfascial intramuscular masses. Histologically, all four tumors were of the typical variant. All patients underwent a R0-surgical resection of the tumor and they were recurrence-free at last follow-up (mean: 47 months; range: 25-87). Conclusion: Hibernoma may present as huge deep intramuscular soft tissue mass in adults, closely mimicking well differentiated liposarcoma and should be considered in the differential diagnosis of fatty soft tissue tumors in any location. Surgical excision is the treatment of choice. The tumor has no malignant or recurrence potential.
Hibernoma; brown tissue; soft tissue tumor; deep-seated
SH2-containing inositol 5’-phosphatase 2 (SHIP2) is a vital regulator of phosphoinositide pools in metabolic pathways and is considered to downregulate phosphatidylinositol 3’-kinase signaling, which underlies the development of several kinds of human cancers. However, SHIP2 expression in non-small cell lung cancer (NSCLC) and its relationship with the clinical characteristics of NSCLC remain poorly understood. In this study, one-step quantitative reverse transcription-polymerase chain reaction and immunohistochemistry analysis with tissue microarray was used to evaluate SHIP2 expression in NSCLC and to investigate the relationship of this expression to NSCLC prognosis. Results showed that the expression of SHIP2 messenger RNA and protein was significantly higher in NSCLC than in corresponding non-cancerous tissues (both p < 0.05). SHIP2 protein expression in NSCLC was related to lymph node metastasis (p = 0.042), TNM stage (p = 0.036), and 5-year survival rate (p = 0.046). The Kaplan-Meier method and log-rank test suggested that high SHIP2 expression, tobacco consumption, and advanced tumor stage were significantly associated with low survival of NSCLC patients. The results of this research suggested that SHIP2 expression was correlated with malignant phenotypes of NSCLC and may thus serve as a poor prognostic factor and valuable oncogene for NSCLC.
SHIP2; NSCLC; qPCR; immunohistochemistry; prognosis
Acute promyelocytic leukemia (APL) has two morphological variants, namely macrogranular (M3) and microgranular (M3v). M3v, characterized by the presence of neoplastic promyelocytes with only sparse fine azurophilic granules, accounts for 10-25% of all APL and has unique biological characteristics. Relapse occurs in approximately 20% of patients with APL. The morphological type of the leukemic cells at relapse is usually identical with the primary disease, and only one case of morphological change at relapse has been reported. Here, we analyzed the clinicopathological features of APL, including 4 relapsed cases emphasizing morphological changes at the time of relapse. The unique finding of the present study is that 2 of 4 relapsed cases changed from M3 to M3v at relapse. The morphological features of these were different in each case (one had blastic features and the other resembled monocytoid leukemic cells). Cytogenetic analyses revealed the continued presence of t(15;17)(q22;q12) at the time of relapse and morphological change. Moreover, the immune phenotype of the leukemic cells changed from CD2-/CD34- to CD2+/CD34+ at that time. These findings suggest that morphological change at relapse in APL may not be a rare event, and that the leukemic cells can show variable morphological features at the time of relapse, which could result in misdiagnosis as a different type of acute myeloid leukemia. Therefore, a comprehensive approach with emphasis on combined morphological, immunophenotypic, and cytogenetic analyses is important for diagnosis and appropriate treatment of relapsed APL.
Acute promyelocytic leukemia; macrogranular variant; microgranular variant; relapse