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目的 探索新克隆的编码人蛋白翻译起始因子的全长基因 (C2基因 )对人胃癌细胞体内、体外成瘤性的影响。方法 将已构建的C2基因真核表达载体 ,与阴性对照的空载体扩增纯化并同时转染人胃癌细胞系SGC790 1细胞 ,获得稳定高表达C2蛋白的人胃癌细胞株 ,应用平板克隆形成实验检测转化细胞的体外成瘤性 ,并在裸鼠体内进行肿瘤细胞体内成瘤性实验 ,了解C2基因转染对人胃癌细胞体内、体外成瘤性的影响。结果 经WesternBlot法和流式细胞术证实 ,C2基因转化细胞稳定高表达C2蛋白。平板克隆形成实验提示 :转染了C2基因和空载体的两种SGC790 1细胞的克隆形成率分别为4 3.8%和 76 .9% (P <0 .0 5 )。裸鼠体内成瘤性实验提示 :转染了C2基因的SGC790 1细胞在裸鼠皮下形成肉眼可见的肿瘤结节所需时间较转染了空载体的SGC790 1细胞长 ,分别为 1周和 2周 ,在相同时间内在裸鼠皮下形成的瘤结节的体积明显小于转染了空载体的细胞在裸鼠皮下形成的瘤结节的体积 ,分别平均为 1.2 0和 5 .5 8cm3 (P <0 .0 1)。结论 C2基因在体内和体外均可抑制人胃癌细胞的成瘤性 ,在肿瘤形成速度和体积方面抑制了SGC790 1细胞的恶性行为 ,提示C2基因可能通过确保蛋白翻译过程的准确性而阻止细胞无限生长 ,因此可能是一个具有抑制细胞增殖功
Objective To explore the effect of the full-length gene (C2 gene) encoding the human protein translation initiation factor of the new clone on human gastric cancer cell in vitro and in vivo. Methods The constructed C2 gene eukaryotic expression vector and empty vector of negative control were amplified and purified and transfected into human gastric cancer cell line SGC790 1 at the same time to obtain stable and high expression of C2 protein human gastric cancer cell line. The in vitro tumorigenicity of the transformed cells was examined. In vivo tumorigenicity experiments of tumor cells in nude mice were conducted to understand the effect of C2 gene transfection on the tumorigenicity of human gastric cancer cells in vitro and in vivo. Results Western Blot and flow cytometry confirmed that the C2 gene transformed cells stably expressed high level of C2 protein. The results of plate clone formation suggested that the clonogenic rates of two SGC7901 cells transfected with C2 gene and empty vector were 43.8% and 76.9%, respectively (P <0.05). In vivo tumorigenicity experiments in nude mice suggest that SGC7901 cells transfected with C2 gene take longer to form macroscopic tumor nodules in nude mice than SGC7901 cells transfected with empty vector for 1 week and 2 The volume of nodules formed in nude mice subcutaneously at the same time was significantly smaller than the volume of nodules formed in nude mice transfected with empty vector, respectively, with an average of 1.2 0 and 5.58 cm 3, respectively (P < 0 .0 1). Conclusion C2 gene can inhibit the tumorigenicity of human gastric cancer cells both in vivo and in vitro, and inhibit the malignant behavior of SGC7901 cells in terms of tumor formation rate and volume, suggesting that C2 gene may prevent cells from infilling by ensuring the accuracy of protein translation process Growth, and therefore may be a work with inhibition of cell proliferation