Preparation of lentivirus silencing PHF8 and its effects on human esophageal squamous-cell carcinoma cells

doi:10.3969/j.issn.1006-7795.2014.01.026

Abstract

Abstract:

Objective To construct a recombinant lentivirus for RNA interference of PHF8 gene, establish a human esophageal squamous-cell carcinoma cell lines with PHF8 gene knockdown and observe its effect on cell proliferation. Methods The PHF8 or Nonsilencing shRNA and packaging vectors were cotransfected to 293T cells to produce the recombinant lentivirus, which were further ultra-centrifuged, tittered and used to infect the human esophageal squamous-cell carcinoma cell lines. The infection efficiency was evaluated with fluorescence microscope and flow cytometry and stable cell lines were constructed using puromycin selection assay. Real-time PCR and Western Blotting were used to confirm the silencing of PHF8. Research on the effect of PHF8 on cell proliferation was performed using MTS asssy. Results We successfully prepared the recombinant lentivirus for RNA interference of PHF8 gene. The expression of PHF8 mRNA and protein in expremental group were lower than that of the control group (P<0.05). The proliferation of expremental group cells were reduced than that of control group (P<0.05). Conclusion Lentivirus mediated shRNA effectively inhibited the expression of PHF8 in human esophageal squamous-cell carcinoma cell lines and the cell proliferation ability.

Key words: Plant homeodomain finger protein 8, lentivirus, esophageal squamous cell carcinoma, cell proliferation

CLC Number:

R735.1

Sun Xiujing, Yuan Biao, Zhu Shengtao, Li Peng, Xu Youqing. Preparation of lentivirus silencing PHF8 and its effects on human esophageal squamous-cell carcinoma cells[J]. Journal of Capital Medical University, 2014, 35(1): 122-128.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2014.01.026

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2014/V35/I1/122

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