BcL2shRNA稳定转染联合γ射线对胃癌SG松C7901细胞凋亡的影响.docxVIP

优秀毕业论文 精品参考文献资料 青岛大学 硕士学位论文 Bcl--2shRNA稳定转染联合γ射线对胃癌SGC--7901细胞凋亡的 影响 姓名：王宁宁 申请学位级别：硕士 专业：肿瘤学 指导教师：沈方臻于两 于两 要 目的：观察Bcl．2shRNA稳定转染联合Y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pGPHl／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，Y线照射 后形成4组细胞分别命名为SGC-7901(A组)，照射／SGC一7901(B组)， Bc卜2shRNA／SGC一7901(C组)，照射／Bcl一2shRNA／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(P0．05)；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±5．27)％，(30．1±6．49)％， (98．4±11．35)％。A、B、C、D组细胞凋亡率分别为(3．80±0．22)％、 (20．8±4．15)％、(23．2±4．34)％、(92．9±25．9)％，差异有统计 学意义(尸O．05)。 结论：Bcl-2基因siRNA干扰联合Y线照射可协同抑制SGC一7901细 胞中Bcl一2的表达，诱导细胞凋亡。 关键词：关键词：胃癌细胞：B细胞淋巴瘤／白血病-2；RNA干扰；细 胞凋亡；照射 ＼煳燃Stable ＼煳燃 Stable expressing shRNA influence bcl-2 gene associating with丫 irradiation On apoptosis of gastric carcinoma cell line SGC一790 1 objective：To observe the effect of bcl-2 gene—specific RNA interference on apotosis and radiosensitivity of gastnc carcinoma cell line SGC7901． Methods：The recombinant eukaryotic expression vertor pGPH 1／GFP／Neo and designed to target bcl-2 gene by lipofectamin mediated gene transfection into SGC790 1．The positive clones were selected with G4 1 8， after Y irradiation．4．group cells were formed：SGC．790 1 ( A group ) ．irradiation／SGC．790 1．(B group ) ，Bcl一2shRNA／SGC-790 1 ( C group ) irradiation／Bel一2shRN～SGC．790 1(D group)．Cell proliferation was detected by CCK一8．Cell apoptotic was observed by AO／PE．The apoptotic rate was determined by flow cytometry．朊stem blot was used to compare the express of protein between transfection cells and non transfection cells． Results：Western blot analyses indicated that the expression of bcl-2 was suppressed by the shRNA and irradiation．The growth of group D was obviously slower than other groups(P0．05)，Cell inhibitory rate of B、C、D groups were(27+5．27)％，(30．1±6．49)％，(98．4±11．35)％。 the apoptosis rate ofA、B、C、D were(3．8±0．22)％．(20．8±4．15) ％， (23．2±4．15)％，(