论文部分内容阅读
目的研究垂体肿瘤转化基因1(Pituitary Tumor Tansforming Gene1,PTTG1)过表达对人结肠癌细胞SW480增殖和周期的影响及其可能机制。方法采用脂质体转染法将pc DNA3.1(+)-PTTG1及空载pc DNA3.1(+)质粒转染人结肠癌SW480细胞,G418法筛选阳性克隆。Western blot法检测PTTG1和cyclin D、cyclin E、p21的表达。MTT法检测细胞增殖,流式细胞术检测细胞周期。结果 (1)成功获得PTTG1高表达的SW480细胞pc DNA3.1(+)-PTTG1及对照pc DNA3.1(+)细胞,Western blot结果显示pc DNA3.1(+)-PTTG1细胞PTTG1表达量分别为pc DNA3.1(+)细胞和SW480细胞的3.58倍和3.42倍;(2)过表达PTTG1基因后,SW480细胞周期改变,G0/G1期细胞减少而S期和G2/M期细胞增加;细胞周期调控蛋白cyclin D和cyclin E表达升高,p21表达降低;SW480细胞增殖显著性加快。(3)过表达PTTG1基因后,SW480细胞中PI3K/AKT信号活化增强,使用PI3K/AKT信号抑制剂LY29400干预后,细胞增殖减弱,周期蛋白cyclin D和cyclin E表达降低,p21表达升高。结论 PTTG1基因过表达可能通过活化PI3K/AKT信号加速SW480细胞从G1期向S期的转化促进细胞增殖,提示PTTG1蛋白可能成为结肠癌治疗的一个潜在靶点。
Objective To investigate the effect of PTTG1 overexpression on proliferation and cell cycle of human colon cancer cell line SW480 and its possible mechanism. Methods pcDNA3.1 (+) - PTTG1 and pcDNA3.1 (+) vector were transfected into human colon cancer SW480 cells by lipofection. The positive clones were screened by G418 method. Western blot was used to detect the expressions of PTTG1, cyclin E and p21. Cell proliferation was detected by MTT assay and cell cycle was detected by flow cytometry. Results (1) The expression of PTTG1 in pcDNA3.1 (+) - PTTG1 cells was successfully detected by pcDNA3.1 (+) - PTTG1 and pcDNA3.1 (+ Which was 3.58-fold and 3.42-fold higher than that of pcDNA3.1 (+) cells and SW480 cells, respectively. (2) The SW480 cell cycle was changed after overexpression of PTTG1 gene, while cells in G0 / G1 phase decreased and cells in S phase and G2 / M phase increased; The expression of cyclin D and cyclin E was increased and the expression of p21 was decreased. The proliferation of SW480 cells was significantly accelerated. (3) Activation of PI3K / AKT signal was enhanced in SW480 cells after overexpression of PTTG1 gene. After PI3K / AKT inhibitor LY29400 was administered, the cell proliferation was weakened, the expressions of cyclin D and cyclin E were decreased and the expression of p21 was increased. Conclusion Overexpression of PTTG1 may promote the proliferation of SW480 cells by activating PI3K / AKT signaling from G1 phase to S phase, suggesting that PTTG1 may be a potential therapeutic target for colon cancer.