ASPP2增强结肠癌HCT116细胞对奥沙利铂的化疗敏感性

The Role of ASPP2 in Chemosensitivity of HCT116 p53^(+/+) Cell Line to Oxaliplatin

为研究ASPP2对奥沙利铂诱导的结肠癌细胞系HCT116 p53+/+(野生型)凋亡及周期的影响.利用ASPP2(rAd-ASPP2)及p53腺病毒(rAd-p53)感染HCT116 p53+/+细胞,经奥沙利铂50μmol/L诱导细胞凋亡及周期改变.Western印迹检测ASPP2及p53的表达水平;MTT法检测ASPP2腺病毒对奥沙利铂诱导的HCT116细胞活性的影响;Calcein/PI吸收试验检测细胞凋亡情况;流式细胞术分析细胞周期分布.结果显示,ASPP2、p53共同过表达,或者ASPP2单独过表达均能增强奥沙利铂诱导的HCT116 p53+/+细胞增殖抑制,以及S期抑制并伴有细胞凋亡水平的升高;而无奥沙利铂诱导时,ASPP2对HCT116 p53+/+细胞的活性、细胞周期及细胞凋亡水平的影响无统计学意义.上述结果表明,ASPP2能够增强奥沙利铂诱导HCT116 p53+/+细胞的增殖抑制、细胞周期抑制和细胞凋亡.

To investigate the role of ASPP2 in apoptosis and cell cycle modulation following oxahplatln treatments in wild type p53＋/＋ HCTll6 colorectal cancer ceils, we used rAd-ASPP2 and rAd-p53 to overexpress ASPP2 and p53 in the presence of 50μmol/L of oxaliplatin for 48 hours. The p53 and ASPP2 expression levels were determined by Western blot; the cell viability were evaluated by MTT; the cellular apoptosis were assayed by calcein/PI staining; and the changes in cell cycle were analyzed by flow cytometry. The results showed that either over expression of ASPP2 alone or together with p53 intensified the inhibition on cell proliferation and S phase percentage in oxaliplatin treated cells. However, the increase of apoptosis was not observed. In the absence of oxaliplatin, the effects of ASPP2 overexpression were statistically insignificant. It was implied that the overexpression of ASPP2 enhanced cell proliferation inhibition, cell cycle regulation, as well as levels of apoptosis under oxaliplatin-induced conditions in HCTll6 p53 ＋/＋ ceils.