摘要
目的探讨10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)对胰腺癌细胞质(ASPC1)血管内皮生长因子(VEGF)蛋白表达、细胞周期和增殖的影响。方法将质粒pEAK8PTEN和pEAK8分别转染指数生长期的胰腺癌细胞株(ASPC1),挑选阳性细胞克隆,扩增培养,用RT PCR、Western印迹、流式细胞术、生长速率等方法分别检测PTEN对ASPC1细胞VEGF蛋白、细胞周期及增殖能力的影响。结果ASPC1细胞转染后,PTENmRNA表达量是转染前的3倍;ASPC1、ASPC1pEAK8、ASPC1pEAK8PTEN细胞PTEN蛋白表达量分别为13.2、12.6和21.6,VEGF蛋白表达量分别为17.2、16.5和13.1,转染后较转染前降低。细胞周期显示,ASPC1pEAK8PTEN细胞较ASPC1、ASPC1pEAK8细胞G2/M、S期细胞增多,G1期细胞减少,并出现少量凋亡细胞(P<0.01)。ASPC1pEAK8PTEN细胞生长曲线平缓,生长速度较另两种细胞明显降低。结论PTEN可使ASPC1细胞VEGF蛋白表达下降,细胞阻滞在G2/M期,抑制肿瘤细胞增殖。
Objective To investigate the effect of phosphatase and tensin homologue deleted on ~chromosome ten (PTEN) on cell cycle, expression of vascular endothelial growth factor (VEGF) and ~proliferation in a human pancreatic cancer cell line (ASPC-1). Methods The pancreatic cancer ASPC-1 cell was transfected with an eukaryotic expression plasmid (pEAK8) inserted PTEN or not in vitro by ~lipofectin methods, and positive cell clones were selected and amplified. Effects of PTEN on ASPC-1 cells were measured by flow cytometry, Western blot and tumor growth curve. Results PTEN mRNA ~expression in the ASPC-1-pEAK8-PTEN cell was two fold up than that in ASPC-1 cells. PTEN and VEGF protein expressions in the ASPC-1, ASPC-1-pEAK8 and ASPC-1-pEAK8-PTEN cells were 13.2, 12.6, 21.6 and 17.2, 16.5, 13.1, respectively. It was found that PTEN protein could be much more ~efficiently expressed in transfected ASPC-1 cell contained plasmid pEAK8-PTEN, whereas the expression of VEGF protein decreased comparing with ASPC-1 or ASPC-1-pEAK8 cell. In addition, in ASPC-1 cells carrying exogenous PTEN, apoptosis cells, and the cells in S and G2/M phase increased whereas G1 phase cells decreased (P<~0.01 ). The growth of cell transfected with PTEN was dramatically ~inhibited compared with the parent ASPC cell or ASPC-1-pEAK8 cell. Conclusion The findings suggest ~that exogenous PTEN inhibit ASPC-1 cell growth by increasing PTEN and decreasing VEGF expression, which led to the ASPC-1 cells in G2/M phase increase.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2005年第3期162-165,共4页
Chinese Journal of Digestion
