论文部分内容阅读
目的观察Bcl-2shRNA稳定转染联合γ线照射对胃癌细胞SGC-7901凋亡的影响。方法构建针对Bcl-2基因的干扰质粒pGPH1/GFP/Neo,经脂质体介导转染SGC-7901细胞,G418筛选稳定表达的细胞株,γ线照射后形成4组细胞,分别命名为SGC-7901(A组)、照射/SGC-7901(B组)、Bcl-2shRNA/SGC-7901(C组)、照射/Bcl-2shRNA/SGC-7901(D组)。CCK-8检测细胞增殖,AO/PE观察细胞凋亡,流式细胞仪检测细胞凋亡,Western-blot测定Bcl-2蛋白表达量的改变。结果 Bcl-2shRNA、照射均可抑制Bcl-2蛋白表达,且二者有协同作用,差异有统计学意义(P<0.05);D组细胞生长慢于B组、C组细胞,B、C、D组细胞增殖抑制率分别为(27.00±5.27)%、(30.10±6.49)%、(98.40±11.35)%。A、B、C、D组细胞凋亡率分别为(3.80±0.22)%、(20.80±4.15)%、(23.20±4.34)%、(92.90±25.90)%,差异有统计学意义(P<0.05)。结论 Bcl-2基因siRNA干扰联合γ线照射可协同抑制SGC-7901细胞中Bcl-2的表达,诱导细胞凋亡。
Objective To observe the effect of stable Bcl-2 shRNA transfection combined with γ-ray irradiation on apoptosis of gastric cancer cell line SGC-7901. Methods Plasmid pGPH1 / GFP / Neo targeting Bcl-2 gene was constructed and transfected into SGC-7901 cells by lipofectamine. Stably expressing cell lines were screened by G418. Four groups of cells were formed after γ-ray irradiation and were named SGC -7901 (Group A), irradiation / SGC-7901 (Group B), Bcl-2 shRNA / SGC-7901 (Group C), irradiation / Bcl-2 shRNA / SGC-7901 (Group D). Cell proliferation was detected by CCK-8, apoptosis was observed by AO / PE, apoptosis was detected by flow cytometry, and the expression of Bcl-2 protein was detected by Western-blot. Results Bcl-2shRNA and Bcl-2showed a significant inhibitory effect on the expression of Bcl-2 protein (P <0.05). The cell growth in group D was slower than that in group B, C, B, The inhibitory rates of cell proliferation in group D were (27.00 ± 5.27)%, (30.10 ± 6.49)%, (98.40 ± 11.35)%, respectively. The apoptotic rates in groups A, B, C and D were (3.80 ± 0.22)%, (20.80 ± 4.15)%, (23.20 ± 4.34)% and (92.90 ± 25.90)%, respectively, with statistical significance 0.05). Conclusion Bcl-2 gene siRNA interference combined with γ-ray irradiation can synergistically inhibit the expression of Bcl-2 in SGC-7901 cells and induce apoptosis.