Survivin shRNA-APC 双基因对结肠癌细胞中 P21 和FHIT 表达的影响

Effects of Survivin shRNA-APC double gene on the expressions of P21 and FHIT in colon cancer cells

摘要:

目的 探究Survivin shRNA-APC 双基因共表达稳转株对HT-29 结肠癌细胞裸鼠皮下移植瘤组织 细胞中P21 和FHIT 表达的影响。方法 选取35 只雌性裸鼠分为双基因组、Survivin shRNA 组、APC 组、空 载组及阴性对照组，每组7 只。培养构建成功的Survivin shRNA-APC 双基因共表达稳转株、Survivin shRNA 稳 转株、APC 稳转株、空载稳转株及HT-29 结肠癌细胞，在每只裸鼠右腋下接种相应的细胞悬液复制移植瘤模型。 测量每组瘤重、计算瘤重抑制率；采用免疫组织化学法测定P21 和FHIT 蛋白的表达。结果 所有裸鼠腋下产 生肿瘤。APC 组、Survivin shRNA 组、双基因组P21、FHIT 蛋白表达量较阴性对照组、空载组升高（P <0.05）， 双基因组的蛋白表达量较APC 组、Survivin shRNA 组升高（P <0.05）。APC 组、Survivin shRNA 组、双基因 组的平均瘤重较阴性对照组和空载组降低（P <0.05）。双基因组平均瘤重较APC 组、Survivin shRNA 组降低 （P <0.05）。APC 组、Survivin shRNA 组、双基因组的瘤重抑制率较空载组升高（P <0.05）。双基因组瘤重抑制 率较APC 组、Survivin shRNA 组升高（P <0.05）。结论 Survivin shRNA-APC 双基因共表达稳转株可能通过上 调P21 和FHIT 蛋白的表达来调节细胞周期，抑制细胞增殖，进而抑制肿瘤生长。其抑制细胞增殖效果比Survivin shRNA、APC 单基因稳转株更显著。

Abstract:

Objective To investigate the effects of co-expression stably transfected cell lines of Survivin shRNA-APC double gene on the expressions of P21 and FHIT in subcutaneous transplanted tumor tissues cell of HT-29 colon cancer in nude mice. Methods A total of 35 female nude mice were randomly divided into five equal groups, namely double-gene group, Survivin shRNA group, APC group, empty vector group, and negative control group. These constructed Survivin shRNA-APC double-gene co-expression stably transfected cell lines, Survivin shRNA stably transfected cell lines, APC stably transfected cell lines, empty vector stably transfected cell lines, and HT-29 colon cancer cells were injected into the right axillary of nude mice respectively to establish SXT models.Tumor weight was measured and the tumor weight inhibitory rate was calculated. P21 and FHIT protein expression were detected by immunohistochemical method. Results Every nude mouse developed tumors. The expressed quantity of protein in the APC group, Survivin shRNA group, and double-gene group, as compared with those in the empty vector group and negative control group, was increased (P < 0.05). The expressed quantity of protein in the double-gene group, as compared with those in the APC group and Survivin shRNA group, was increased (P < 0.05). The average tumor weight in the APC group, Survivin shRNA group, and double-gene group, as compared with those of empty vector group and negative control group, was reduced (P < 0.05). The average tumor weight in the doublegene group, as compared with those in the APC group and Survivin shRNA group, was reduced (P < 0.05). The tumor weight inhibition rate in the APC group, Survivin shRNA group, and double-gene group, as compared with those of empty vector group, was increased (P < 0.05). The tumor weight inhibition rate in the double-gene group, as compared with APC group and Survivin shRNA group, was increased (P < 0.05). Conclusions co-expression stably transfected cell lines of Survivin shRNA-APC double gene can adjust cell cycle, inhibit cell proliferation, and inhibit tumor growth by upregulating expression of P21 and FHIT. Its effects of inhibiting were more noticeable than those of Survivin shRNA or APC single gene stable strain.