Cotransfection of the p15INK4B and p21WAF1 genes synergistically inhibits proliferation and promotes apoptosis in human esophageal Squamous cell carcinoma cell line EC109
- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 30
- DOI:
- 10.3969/j.issn.1009-3079.2009.30.002
- Key Word:
- p15INK4B;P21WAF1;基因转染;食管鳞癌
Abstract： 目的:探讨p15INK4B和p21WAF1基因联合转染对人食管鳞癌细胞系EC109细胞增殖和凋亡的影响.方法:脂质体介导PcDNA3.1(+)-p15和pcDNA3.1(+)-p21转染EC109细胞,稳定筛选后用RT-PCR检测转染细胞p15与p21基因mRNA表达,Western blot检测转染细胞P15和P21蛋白的表达.用MTT法和透射电镜检测p15和p21基因分别及联合转染对EC109细胞增殖与凋亡的影响,流式细胞仪检测EC109细胞周期分布与凋亡率.结果:p15和p21转染组EC109细胞生长速度低于空载体组与未转染组,联合转染组与二者单独转染组相比.亦明显抑制EC109细胞体外生长速度.p15和p21转染组EC109细胞发生G1/S阻滞,G1期细胞比例显著高于空载体组和未转染组,S期则显著降低(G1期:60.52%±3.75%,63.12%±2.89% vs 42.17%±5.30%.41.38%±6.54%;S期:22.67%±1.25%,17.96%±2.03% vs 30.96%±3.33%,36.05%±1.78%,均P<0.01),并出现凋亡峰,透射电镜亦发现p15和p21转染组发生细胞凋亡,联合转染组发生更为明显的G1/S阻滞,G1期比例显著升高、S期比例明显降低(G1期:72.83%±2.31% vs60.52%±3.75%,63.12%±2.89%:S期:13.59%±2.59% vs 22.67%±1.25%,17.96%±2.03%.均P<0.05),凋亡率明显升高(21.21%±1.78%vs 4.32±1.74%,10.83%±2.40%,均P<0.01).结论:p15和p21基因联合转染在体外可以进一步增强对人食管鳞癌EC109细胞的抑制与诱导凋亡作用.
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