论文部分内容阅读
目的:构建过表达血清与糖皮质激素调节激酶3(SGK3)的质粒以及稳定表达SGK3的肝癌细胞系BEL-7402,研究其在去甾体激素胎牛血清(FBS)中的抗凋亡能力。方法:PCR扩增SGK3基因,将扩增产物连接到p CDH载体,构建出p CDH-SGK3的慢病毒载体质粒,将其同空白对照p CDH-NC分别与慢病毒包装载体共转入293T细胞,包装成慢病毒p CHD-SGK3和p CDH-NC;将构建的慢病毒感染肝癌细胞BEL-7402并用嘌呤霉素筛选,Western印迹检测SGK3的表达;观察细胞在FBS及去甾体激素FBS中的生长情况。结果:包装出p CDH-SGK3重组慢病毒,此慢病毒感染肝癌细胞系BEL-7402后获得表达;CCK8实验表明过表达SGK3可促进肝癌细胞的生长,BEL-7042细胞中有雄激素的表达,去甾体激素FBS中细胞生长受到抑制,过表达SGK3可增强肝癌细胞在去甾体激素血清中的抗凋亡能力。结论:在肝癌细胞BEL-7402中过表达SGK3可促进细胞生长,可增强细胞在去甾体激素FBS中的抗凋亡能力。
OBJECTIVE: To construct a plasmid that overexpresses serum and glucocorticoid-regulated kinase 3 (SGK3) and a hepatocellular carcinoma cell line BEL-7402 stably expressing SGK3 to investigate its anti-apoptotic capacity in steroid-deprived fetal bovine serum (FBS). Methods: The SGK3 gene was amplified by PCR. The amplified product was ligated into pCDH vector to construct lentiviral plasmid pCDH-SGK3. The recombinant plasmid pCDH-NC was transfected into 293T cells with lentivirus vector And packaged into lentivirus p CHD-SGK3 and p CDH-NC. The constructed lentivirus was used to infect hepatoma BEL-7402 cells and screened with puromycin. The expression of SGK3 was detected by Western blotting. The expression of SGK3 in FBS and steroid- The growth of the situation. Results: The pCDH-SGK3 recombinant lentivirus was packaged and the lentivirus infected the hepatocellular carcinoma cell line BEL-7402. The results of CCK8 assay showed that over-expression of SGK3 could promote the growth of hepatoma cells and the expression of androgen in BEL-7042 cells. To steroid hormone FBS cell growth was inhibited, overexpression of SGK3 can enhance the anti-apoptotic ability of liver cancer cells in steroid-resistant serum. Conclusion: Overexpression of SGK3 in hepatoma BEL-7402 cells can promote cell growth and enhance the anti-apoptotic ability of cells in steroid-resistant FBS.