Bcl-2shRNA稳定转染联合γ射线对胃癌SGC-7901细胞凋亡的影响

Influence of stable transfection of bcl-2shRNA combined with γ irradiation on apoptosis of the gastric carcinoma cell line SGC-7901

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 bcl-2 gene-specific RNA interference combined with γ irradiation on apoptosis and radiosensitivity of the gastric carcinoma cell line SGC7901.Methods The recombinant eukaryotic expression vector pGPH1/GFP/Neo designed to target the bcl-2 gene was transfected into SGC7901 cells by lipofectamin.Stable positive clones were selected with G418.After γ irradiation,the cells were divided into 4 groups： SGC-7901（group A）,irradiation/SGC-7901（group B）,Bcl-2shRNA/SGC-7901（group C） and irradiation/Bcl-2shRNA/SGC-7901（group D）.Cell proliferation was detected by CCK-8.Cell apoptosis was observed by AO/PE.The apoptosis rate was determined by flow cytometry.Expression of bcl-2 was determined by Western blot,and compared between transfection cells and non-transfection cells.Results Western blot analysis indicated that expression of bcl-2 was suppressed by shRNA and irradiation,and they had a synergetic effect.The growth of cells in group D was obviously slower than that in the other groups（P 〈0.05）.Cell inhibitory rates in groups B,C and D were（27.00±5.27）%,（30.10±6.49）% and（98.40±11.35）%,respectively.Apoptosis rates in groups A,B,C and D were（3.80±0.22）%,（20.80±4.15）%,（23.20±4.15）% and（92.90±25.90）%,respectively（P 〈0.05）.Conclusions Bcl-2 gene suppression by RNAi and irradiation can induce cell apoptosis,co-inhibit expression of bcl-2,and obviously increase radiosensitivity in the human gastric cell line. 更多还原