卵巢癌肿瘤干细胞对顺铂和盐霉素的药物敏感性

DRUG SENSITIVITY OF HUMAN OVARIAN CANCER STEM CELLS TO CISPLATIN AND SALINOMYCIN

目的检测卵巢癌干细胞对盐霉素、顺铂的药物敏感性,并比较两者对卵巢癌干细胞的杀伤作用。方法采用无血清悬浮培养法从人卵巢癌细胞株SKOV-3中分离肿瘤干细胞,并采用流式细胞技术检测干细胞标志性因子CD44。人卵巢癌SKOV-3细胞及人卵巢癌干细胞经盐霉素、顺铂作用48h后,采用CCK-8比色法检测细胞存活率,计算半数致死量(IC50)。取第48小时时间点的半数抑制浓度作用于悬浮培养的第5代悬浮细胞48h后,收集药物生存细胞(DSC),采用RT-PCR技术检测DSC的三磷酸腺苷结合转运蛋白B1(ABCB1)基因及三磷酸腺苷结合转运蛋白G2(ABCBG2)基因的转录水平。结果无血清悬浮培养法富集的人卵巢癌微细胞球高表达CD44抗原。盐霉素、顺铂作用第48小时时间点的卵巢癌SKOV-3细胞的IC50分别为(62.50±2.53)和(3.62±0.38)μmol/L,卵巢癌干细胞IC50分别为(48.81±2.83)和(3.80±0.22)μmol/L。卵巢癌干细胞对顺铂耐药指数(RI)为2.12±0.41,明显高于盐霉素RI(0.78±0.08),两者差异有显著意义(t=6.22,P<0.05)。盐霉素组DSC细胞ABCB1和ABCBG2基因表达水平明显低于顺铂组(t=7.49、9.75,P<0.05)。结论人卵巢癌干细胞对盐霉素药物敏感性显著高于顺铂。盐霉素对卵巢癌干细胞的杀伤作用大于顺铂。该差异可能与干细胞高表达ABCB1、ABCBG2基因有关。

Objective To detect drug sensitivity of human ovarian tumor stem cells to cisplatin and salinomycin and com- pare the effect of the two drugs on ovarian cancer stem cells （OCSCs）. Methods Isolating and identifying tumor stem cells from SKOV-3 strains of OCSCs, the marker factor CD44 was measured by flow cytometry. After 48 hours of treatment with salinomy- cin and cisplatin, the cell survival rate and median lethal dose （MLD 50） were determined by cell counting kit-8 （CCK-8）. ABCB1 and ABCBG2 mRNA were tested by RT-PCR in the experimental group and control group at 48-hour time point. Results CD44 antigen expression was high in OCSCs. The MLD 50 of salinomycin and cisplatin to ovarian cancer cell SKOV-3 was （62.50±2.53） and （3.62±0.38） μmol/L, and that to OCSCs was （48.81±2.83） and （3.80±0.22）μmol/L, respectively. The drug re sistance index （RI） of OCSCs to cisplatin was higher than that to salinomycin being 2.12 ± 0.41 versus 0.78 ± 0.08 （t = 6.22, P〈 0.05）. The expressions of ABCB1 and ABCBG2 in OCSCs salinomyein group was obviously lower than that in cisplatin group （t= 7. 49,9. 75,P〈0.05）. Conclusion The sensitivity of OCSCs to salinomycin is higher than to cisplatin. The lethal effect of salinomycin on OCSCs is stronger than cisplatin. This contrast between the two drugs is probably associated with the high expressions of both ABCB1 and ABCG2 genes in cancer stem cells.