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目的:研究胃癌相关基因GCRG213正义、反义转染对胃癌细胞MKN45成瘤性的影响。方法:采用半定量RT-PCR及Western免疫印迹法比较转染不同质粒的MKN45细胞中GCRG213在mRNA和蛋白质水平上的表达差异。选取稳定转染不同质粒的MKN45细胞,绘制细胞生长曲线,平板克隆形成实验、裸鼠移植瘤实验分析转染细胞成瘤性。结果:平板克隆形成实验显示转染了GCRG213正向克隆的MKN45细胞其细胞克隆形成数量高于转染空载体的细胞,而转染了GCRG213反向克隆的MKN45细胞其细胞克隆形成数量低于转染空载体的细胞。在裸鼠体内进行的转染细胞的成瘤性实验可见,转染了GCRG213正向克隆的MKN45细胞在裸鼠体内成瘤性高于转染空载体的细胞,而转染了GCRG213反向克隆的MKN45细胞在裸鼠体内成瘤性低于转染空载体的细胞。结论:胃癌相关基因GCRG213可促进肿瘤细胞的生长,促进肿瘤细胞的成瘤性,GCRG213反义转染可抑制肿瘤细胞的生长,抑制肿瘤细胞的成瘤性。
Objective: To investigate the effect of GCRG213 sense and antisense transfection on the tumorigenesis of gastric cancer cell line MKN45. Methods: The mRNA and protein levels of GCRG213 in MKN45 cells transfected with different plasmids were compared by semi-quantitative RT-PCR and Western blotting. MKN45 cells stably transfected with different plasmids were selected for cell growth curve, plate clone formation assay and transplanted tumorigenicity in nude mice. Results: The results of plate clone formation assay showed that the number of clone formation in MKN45 cells transfected with GCRG213 positive clones was higher than that in empty vector transfected cells, while the number of clone formation in MKN45 cells transfected with GCRG213 reverse clone was lower than that of transfected MKN45 cells Empty vector-bearing cells. The tumorigenicity experiment of transfected cells in nude mice shows that the positive clonality of MKN45 cells transfected with GCRG213 in nude mice is higher than that of transfected empty vector and transfected with GCRG213 reverse cloning Of MKN45 cells had less tumorigenicity in nude mice than cells transfected with empty vector. Conclusion: GCRG213 can promote the growth of tumor cells and promote the tumorigenicity of tumor cells. Antisense transfection of GCRG213 can inhibit the growth of tumor cells and inhibit the tumorigenicity of tumor cells.