论文部分内容阅读
目的研究人tRNA甲基转移酶9样蛋白KIAA1456的高表达对HO8910PM细胞增殖及细胞周期的影响。方法利用PCR扩增KIAA1456编码区片段,克隆插入载体Ubi-MCS-EGFP-IRES-puromycin中,构建Ubi-KIAA1456-EGFP-puromycin(LV-KIAA1456)慢病毒表达载体;将慢病毒表达载体包装生成慢病毒颗粒并感染HO8910PM细胞,72 h后观察绿色荧光蛋白(GFP)在HO8910PM卵巢癌细胞中的表达分布情况;反转录PCR检测HO8910PM细胞中KIAA1456 mRNA水平,Western blot法检测HO8910PM细胞KIAA1456蛋白表达水平;CCK-8法检测细胞增殖活性,流式细胞术检测细胞周期。结果成功构建重组慢病毒表达载体LV-KIAA1456,慢病毒颗粒感染HO8910PM细胞后,细胞中KIAA1456的mRNA和蛋白均高表达。过表达KIAA1456的HO8910 PM细胞增殖能力明显抑制、G1期细胞明显增加。结论过表达KIAA1456显著抑制HO8910 PM卵巢癌细胞增殖,细胞周期阻滞于G1期。
Objective To investigate the effect of high expression of human tRNA methyltransferase 9-like protein KIAA1456 on the proliferation and cell cycle of HO8910PM cells. Methods The fragment of KIAA1456 coding region was amplified by PCR and inserted into vector Ubi-MCS-EGFP-IRES-puromycin to construct the lentivirus expression vector of Ubi-KIAA1456-EGFP-puromycin (LV-KIAA1456) HO8910PM cells were infected with virus particles and infected with HO8910PM cells. The expression and distribution of green fluorescent protein (GFP) in HO8910PM ovarian cancer cells were observed 72 hours later. The KIAA1456 mRNA level in HO8910PM cells was detected by reverse transcription PCR. The cell proliferation activity was detected by CCK-8 assay and the cell cycle was detected by flow cytometry. Results The recombinant lentiviral vector LV-KIAA1456 was constructed successfully. After lentivirus particles infected HO8910PM cells, the expression of KIAA1456 mRNA and protein in the cells was high. The proliferation of HO8910 PM cells overexpressing KIAA1456 was significantly inhibited, and the cells in G1 phase were significantly increased. Conclusion Overexpression of KIAA1456 significantly inhibited HO8910 PM ovarian cancer cell proliferation, cell cycle arrest in G1 phase.