论文部分内容阅读
目的研究人端粒保护蛋白(human protection 0f telomeres,hPOTl)对胃癌细胞生长增殖的作用。方法利用本课题组先前所构建的hPOTl正、反义核酸真核表达载体,转染SGC7901胃癌细胞,G418筛选阳性克隆,用Western印迹法、MTT法、流式细胞仪、透射电镜等技术观察基因转染后SGC7901细胞hPOTl蛋白的表达、细胞生长曲线、细胞周期、细胞凋亡及细胞形态学的变化。结果转染了hPOTl反义核酸真核表达载体(pcDNA-as—hPOTl)的SGC7901胃癌细胞hPOTl表达降低,生长减慢,阻滞于G2/M期,细胞凋亡增加。结论hPOTl反义核酸真核表达载体能显著抑制hPOTl蛋白的表达,hPOTl蛋白可能参与胃癌细胞的周期调控,并与细胞的生长增殖有关。
Objective To study the effect of human protection 0f telomeres (hPOT1) on the growth and proliferation of gastric cancer cells. Methods The positive and negative antisense hPOTl eukaryotic expression vectors constructed by our group were used to transfect SGC7901 gastric cancer cells. Positive clones were screened by G418, and the genes were observed by Western blotting, MTT, flow cytometry and transmission electron microscopy The expression of hPOT1 protein, cell growth curve, cell cycle, apoptosis and cell morphological changes of SGC7901 cells after transfection. Results The hPOT1 expression in SGC7901 gastric cancer cells transfected with hPOT1 antisense nucleic acid eukaryotic expression vector (pcDNA-as-hPOT1) decreased, the growth slowed down and arrested in G2 / M phase, and the apoptosis increased. Conclusion hPOTl antisense nucleic acid eukaryotic expression vector can significantly inhibit the expression of hPOTl protein, hPOTl protein may be involved in the cycle of gastric cancer cells, and cell proliferation and proliferation.