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Article Open Access

HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect

  • Authors:
    • Yong‑Fang Li
    • Yang‑Yang Yuan
    • Ying‑Min Zhang
    • Na Zhao
    • Qi Zhang
    • Fan‑Xiu Meng
    • Ran‑Peng Gao
    • Bao‑Feng Yu
    • Yue‑Hong Zhang
    • Rui Guo
    • Hai‑Long Wang
    • Jun Xie
    • Jun Xu
    • Qin Qin
    • Xiu‑Shan Dong
  • View Affiliations / Copyright

    Affiliations: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of General Surgery, Affiliated Tumor Hospital of Shanxi Medical University, Taiyuan, Shanxi 030013, P.R. China, Central Laboratory, Shanxi Provincial People's Hospital, Shanxi Medical University, Taiyuan, Shanxi 030012, P.R. China, Department of General Surgery, Shanxi Dayi Hospital, Taiyuan, Shanxi 030032, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 764-772
    |
    Published online on: May 31, 2017
       https://doi.org/10.3892/mmr.2017.6657
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Abstract

Previous studies demonstrated that herpes simplex virus thymidine kinase (HSVtk) could phosphorylate non‑toxic gancyclovir (GCV) efficiently to produce phosphorylated products that result in cell apoptosis, to kill tumor cells. The present study aimed to construct a plasmid vector, pcDNA3.1‑pAFP‑TK, carrying the suicide gene driven by the alpha‑fetoprotein (AFP) promoter, to investigate the cytotoxicity of HSVtk/GCV suicide gene system on hepatoma carcinoma cells. Reverse transcription‑polymerase chain reaction and western blotting results demonstrated that the HSVtk gene was effectively expressed in HepG2 hepatoma carcinoma cells transfected with pcDNA3.1‑pAFP‑TK plasmid, whereas HSVtk gene expression was not detected in normal HL‑7702 liver cells. In addition, MTT assays indicated that cell viability of HepG2 cells with the plasmid pcDNA3.1‑pAFP‑TK decreased in a dose‑dependent manner following treatment with GCV for 48 h. Flow cytometry also revealed that the cell apoptosis rate and mitochondrial membrane potential reduction rate in the HepG2 cells treated with HSVtk/GCV suicide gene system were significantly higher than in the control group. Apoptosis rates in the control group and the pcDNA3.1‑pAFP‑TK group were (1.00±0.62%) and (38.70±6.03%), respectively. Mitochondrial membrane potential reduction rates in the control group and the pcDNA3.1-pAFP-TK group were (0.57±0.11%) and (22.84±5.79%), respectively. Caspase‑3 staining demonstrated that activated caspase‑3 increased significantly in the HepG2 cells treated with HSVtk/GCV suicide gene system, whereas in the control group activated caspase‑3 increase was not observed. The results of the present study, therefore, indicated that HSVtk suicide gene was obviously expressed in the HepG2 cells and that the HSVtk/GCV system was effective at killing HepG2 hepatoma carcinoma cells.
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Copy and paste a formatted citation
Spandidos Publications style
Li YF, Yuan YY, Zhang YM, Zhao N, Zhang Q, Meng FX, Gao RP, Yu BF, Zhang YH, Guo R, Guo R, et al: HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect. Mol Med Rep 16: 764-772, 2017.
APA
Li, Y., Yuan, Y., Zhang, Y., Zhao, N., Zhang, Q., Meng, F. ... Dong, X. (2017). HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect. Molecular Medicine Reports, 16, 764-772. https://doi.org/10.3892/mmr.2017.6657
MLA
Li, Y., Yuan, Y., Zhang, Y., Zhao, N., Zhang, Q., Meng, F., Gao, R., Yu, B., Zhang, Y., Guo, R., Wang, H., Xie, J., Xu, J., Qin, Q., Dong, X."HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect". Molecular Medicine Reports 16.1 (2017): 764-772.
Chicago
Li, Y., Yuan, Y., Zhang, Y., Zhao, N., Zhang, Q., Meng, F., Gao, R., Yu, B., Zhang, Y., Guo, R., Wang, H., Xie, J., Xu, J., Qin, Q., Dong, X."HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect". Molecular Medicine Reports 16, no. 1 (2017): 764-772. https://doi.org/10.3892/mmr.2017.6657
Copy and paste a formatted citation
x
Spandidos Publications style
Li YF, Yuan YY, Zhang YM, Zhao N, Zhang Q, Meng FX, Gao RP, Yu BF, Zhang YH, Guo R, Guo R, et al: HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect. Mol Med Rep 16: 764-772, 2017.
APA
Li, Y., Yuan, Y., Zhang, Y., Zhao, N., Zhang, Q., Meng, F. ... Dong, X. (2017). HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect. Molecular Medicine Reports, 16, 764-772. https://doi.org/10.3892/mmr.2017.6657
MLA
Li, Y., Yuan, Y., Zhang, Y., Zhao, N., Zhang, Q., Meng, F., Gao, R., Yu, B., Zhang, Y., Guo, R., Wang, H., Xie, J., Xu, J., Qin, Q., Dong, X."HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect". Molecular Medicine Reports 16.1 (2017): 764-772.
Chicago
Li, Y., Yuan, Y., Zhang, Y., Zhao, N., Zhang, Q., Meng, F., Gao, R., Yu, B., Zhang, Y., Guo, R., Wang, H., Xie, J., Xu, J., Qin, Q., Dong, X."HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1‑pAFP-TK and targeted killing effect". Molecular Medicine Reports 16, no. 1 (2017): 764-772. https://doi.org/10.3892/mmr.2017.6657
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