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1.  Function and expression of prolyl hydroxylase 3 in cancers 
Hypoxia inducible factor (HIF) is a product of tumor cells that plays an important role in protecting tumor cells and adjusting to low oxygen tension through driving the progression and aggressiveness of tumors and changing the growth, angiogenesis, differentiation and metastasis of tumors. Prolyl hydroxylase 3 (PHD3) is a member of PHDs that are induced in hypoxia. Many studies have shown that PHD3 not only can hydroxylate HIF-1α, but also has various other biological functions. Thus PHD3 plays significant roles in suppressing the growth, angiogenesis, differentiation and metastasis of tumors and promoting apoptosis of tumors under hypoxic conditions. It may become a new tumor suppressor gene and also may become a new approach to investigate tumors.
PMCID: PMC3776186  PMID: 24049515
prolyl hydroxylase 3; cancer; hypoxia inducible factors
2.  Effect of TSLC1 gene on growth and apoptosis in human esophageal carcinoma Eca109 cells 
To explore the effect of tumor suppressor in lung cancer 1 (TSLC1) on proliferation and apoptosis in esophageal cancer Eca109 cells.
Material and methods
Eca109 cells were divided into three groups: TSLC1 transfected group (TTG), mock group (MG) and untransfected group (UTG). The TTG and MG were transfected transiently with the pIRES2-EGFP-TSLC1 eukaryotic expression vector and pIRES2-EGFP vector respectively. The UTG was a blank control. The TSLC1 expression in TTG was analyzed with the fluorogram and RT-PCR method. Cell proliferation was measured with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. Cell cycle was measured by flow cytometry (FCM). Cell apoptosis was detected by Annexin-V/PI double staining FCM.
Green color was found in TTG and MG. The band of TSLC1 mRNA of TTG was located at about 1400 bp by RT-PCR and agarose gel electrophoresis assay. The TSLC1 inhibited cell proliferation significantly in MTT assay, and the cell proliferation was slower in TTG than MG and UTG. After TSLC1 transfection, cell numbers increased in G0/G1 phase and decreased in S phase. Forty-eight hours after transfection, the apoptosis rate and death rate of TTG were higher than MG and UTG. Thus TSLC1 induced Eca109 cells to apoptosis.
The TSLC1 gene had a potent effect on cell proliferation inhibition, G1/S cell cycle arrest and induction of cell apoptosis in Eca109 cells.
PMCID: PMC3542483  PMID: 23319971
esophageal carcinoma; TSLC1 gene; transient transfection; cell cycle; apoptosis
3.  Construction of a recombinant eukaryotic expression vector containing PHD3 gene and its expression in HepG2 cells 
Prolyl hydroxylase domain 3 (PHD3) is a hypoxia inducible factor-α (HIFα) regulator; it degrades HIFα in the presence of oxygen. Recently, there have been an increasing number of studies about the role of PHD3 in proliferation and apoptosis of cancer cells. However, most of the evidence for the role of PHD3 is observational, and little is known of the molecular mechanism. In our current study, we constructed a recombinant eukaryotic expression vector containing the PHD3 gene and detected its biological activity in human hepatoma cell line (HepG2 cells). We successfully constructed a recombinant pcDNA 3.1(+)-PHD3 plasmid; the results showed that PHD3 overexpression could inhibit the proliferation of HepG2 cells and induce apoptosis by activating caspase-3 activity. Our study has provided preliminary materials and data for further investigation of the effect of PHD3 on HepG2 cells.
PMCID: PMC3511251  PMID: 22898032
Prolyl hydroxylase domain 3 (PHD3); Hepatocellular cancer (HCC); Hypoxia inducible factor (HIF); Caspase-3
5.  Construction of eukaryotic expression vector of TSLC1 gene 
To construct a eukaryotic expression vector of the tumour suppressor in lung cancer 1 (TSLC1) gene, so as to explore the mechanisms of tumour suppression of the gene theoretically.
Material and methods
The open reading frame (ORF) of TSLC1 gene was amplified with RT-PCR from normal human foreskin acrobystia, and cloned to pMD19-T simple vector (TA Clone method). The resultant plasmid was transformed into Escherichia coli JM109 for amplification. The TA Clone recombinant was digested by double restriction enzyme (Bgl II/EcoR I) and analysed with agarose gel electrophoresis. The positive one was sequenced. The inserted DNA fragment was recovered, and then it was mounted into the eukaryotic expression vector pIRES2-EGFP, transformed into E. coli JM109 for amplification. A positive recombinant plasmid named pIRES2-EGFP-TSLC1 was confirmed by Bgl II/EcoR I double-enzyme digestion analysis.
RT-PCR amplified the ORF of the TSLC1 gene. It was approximately 1400 base pairs. The obtained DNA was confirmed a high degree of homology with the sequence of TSLC1 cDNA sequence (AY358334) stored at GenBank.
Construction of a TSLC1 eukaryotic expression vector was successful, and it has established a solid foundation for further study.
PMCID: PMC3258782  PMID: 22291791
TSLC1 gene; construction; eukaryotic expression vector
6.  Prognostic value of serum soluble Fas in patients with locally advanced unresectable rectal cancer receiving concurrent chemoradiotherapy 
Objective: This study was designed to detect the changes of serum soluble Fas (sFas) levels in patients with locally advanced unresectable rectal cancer (LAURC), and to explore its prognostic value of response. Methods: Soluble samples were obtained from LAURC subjects, treated by concurrent chemoradiotherapy, before treatment and one month after treatment. Healthy donor serum samples were used as controls. sFas concentration was measured by enzyme-linked immunosorbent assay (ELISA). Results: The sFas levels before treatment and one month after treatment were both significantly higher in LAURC subjects than in healthy controls [(8.79±1.39) and (7.74±1.32) vs. (5.53±1.13) ng/L, P<0.01]. The sFas levels before treatment and one month after treatment were significantly lower in the response group (complete and partial responses) than in the non-response group (stable and progressive diseases) [(8.50±1.25) vs. (10.17±1.26) ng/L, P<0.01 and (7.50±1.24) vs. (8.90±1.13) ng/L, P<0.01, respectively]. The one-year survival rate was 54.2% and 82.6% in those with sFas levels >8.79 ng/L and <8.79 ng/L before treatment (P<0.02), respectively, 50.0% and 87.0% in those with sFas levels >7.74 ng/L and <7.74 ng/L one month after treatment (P<0.01), respectively. Conclusions: The sFas level is higher in LAURC subjects than in healthy controls. Concurrent chemoradiotherapy can reduce sFas levels in LAURC patients. The monitoring of sFas may provide prognostic information for LAURC patients.
PMCID: PMC2997398  PMID: 21121068
Soluble Fas (sFas); Rectal cancer; Concurrent chemoradiotherapy; Prognosis

Results 1-6 (6)