Akt/mTOR通路相关蛋白在卵巢癌顺铂耐药细胞株SKOV3/DDP中的表达研究  

Establishment of cisplatin-resistant ovarian cancer cell line SKOV3/cDDP and investigation of the expression of Akt/mTOR signaling pathway protein using this cell line

作　　者：徐国才[1] 相媛媛[2] 张莘[1] 林仲秋[1] 张丙忠[1] 

机构地区：[1]中山大学孙逸仙纪念医院妇产科,广州510120 [2]中山市人民医院,中山528400

出　　处：《现代妇产科进展》2013年第2期102-105,共4页Progress in Obstetrics and Gynecology

摘　　要：目的:用大剂量冲击诱导法建立卵巢癌顺铂耐药细胞株SKOV3/DDP。通过检测Akt/mTOR通路相关蛋白在SKOV3/DDP细胞株中的表达,以探讨顺铂耐药的可能机制。方法:采用顺铂20μg/ml,反复间歇24h冲击诱导法建立卵巢癌顺铂耐药细胞株SKOV3/DDP,验证其耐药指数(RI)及生物学指标。Western blot法检测SKOV3/DDP、SK-OV3细胞中Akt/mTOR通路相关蛋白p-Akt、mTOR、p70S6K的表达。结果:(1)SKOV3/DDP细胞耐药指数(RI)为3.079,较SKOV3细胞升高约3倍;(2)SKOV3/DDP细胞的生长速度较SKOV3细胞明显减慢;SKOV3/DDP与SKOV3的G0～G1期细胞所占比例分别为59.02%,44.32%,两者比较差异显著(P=0.029),其中前者出现G0～G1期阻滞;(3)Western blot检测结果显示:①无顺铂作用下,SKOV3/DDP与SKOV3细胞相比,p-Akt蛋白水平升高,mTOR、p70S6K水平下降。②30μg/ml顺铂作用后,p-Akt蛋白表达水平在SKOV3细胞中显著升高,而在SKOV3/DDP细胞中变化不明显;p-mTOR蛋白水平在两者中均有上调趋势,但在SKOV3/DDP细胞中变化较显著;p70S6K蛋白水平亦在两者中均有上调趋势,但趋势不显著,SKOV3/DDP细胞的总体水平低于SKOV3细胞。结论:大剂量冲击诱导方式可以成功建立卵巢癌顺铂耐药细胞株SKOV3/DDP;Akt/mTOR通路与顺铂耐药细胞株形成有关。Objective：To establish the cisplatin-resistant ovarian cancer cell line SK- OV3/DDP by exposing ovarian cancer cell line SKOV3 intervally and repeatedly to high-level concentration of cisplatin, and investigate the mechanism of cisplatin resistance by detecting the expression of Akt/mTOR signaling pathway protein in SKOV3/DDP. Methods： The cisplatin-re- sistant ovarian cancer cell line SKOV3/DDP was established by exposing SKOV3 intervally and repeatedly to high-level concentration of 20p^g/ml cisplatin for 24 hours each time. The biologi- cal features and drug resistance index （RI） were detected. The expressions of p-Akt, roTOR, p70S6K were detected in SKOV3/DDP and SKOV3 by Westem blot. Results： （ 1 ） The value of resistance index （RI） of SKOV3/DDP was elevated more than 3 times, the number was 3. 079. （2）The resistant cell line SKOV3/DDP grew more slowly than SKOV3 cell line （P〈0.05）. The percentage of SKOV3/DDP cells at G0/G1 phase were more than SKOV3 cells （59.02% vs 44.32% , P = 0. 029 ）. The SKOV3/DDP cells were arrested at G0/G1 phase. （ 3 ） Western blot results showed the level of p-Akt was elevated while the level of roTOR and p70S6K were depressed in SKOV3/DDP cells when the cell lines were cultured without cisplatin. While cul- tured with 30μg/ml eisplatin,the level of p-Akt was elevated remarkably in SKOV3 and the level of roTOR was elevated significantly in SKOV3/DDP. The levels of p70S6K were elevated slowlv in both cell lines. Conclusions：The cisplatin-resistant ovarian cancer cell line SKOV3/ DDP can he established by interval and repeated exposing to high-level concentration of cispla- tin successfully. The activation of the Akl/mTOR pathway play an important role in cisplatin re- sistance and the expression of roTOR can be effeeted by many other factors.

关 键 词：信号传导 卵巢肿瘤 抗药性 肿瘤 细胞周期 哺乳类雷帕霉素靶蛋白(mTOR) 

分 类 号：R737.31[医药卫生—肿瘤]