CAJ | 학술논문

目的观察肿瘤干细胞在人骨肉瘤细胞株对顺铂(CDDP)产生获得性耐药中的作用.方法采用大剂量冲击结合逐步增加剂量(1、3、5 mg/L依次递增)的方法诱导产生人骨肉瘤耐CDDP细胞株,0.20 mg/L的As2O3逆转耐药株的耐药性,噻唑蓝(MTT)法和实时定量聚合酶链反应(Real-time PCR)分别检测细胞株对CDDP的敏感性和let-7、微小RNA(miR)-200c以及性别决定区Y框蛋白-2(SOX-2)的表达.结果耐药株的耐药性显著高于亲本株(P＜0.01).As2O3降低了耐药株对CDDP的耐药性,逆转倍数为3.39,相对逆转效率为76.6％.相对于亲本株,耐药株细胞let-7和miR-200c的表达显著降低(P＜0.05),SOX-2的表达显著升高(P＜0.01).As2 03处理后,耐药株细胞let-7和miR-200c的表达显著增加(P＜0.05),SOX-2的表达显著下降(P＜0.05).结论 CSC有可能是人骨肉瘤细胞株MG63对CDDP产生耐药的机制之一.
목적 관찰종류간세포재인골육류세포주대순박(CDDP)산생획득성내약중적작용.방법 채용대제량충격결합축보증가제량(1、3、5 mg/L의차체증)적방법유도산생인골육류내CDDP세포주,0.20 mg/L적As2O3역전내약주적내약성,새서람(MTT)법화실시정량취합매련반응(Real-time PCR)분별검측세포주대CDDP적민감성화let-7、미소RNA(miR)-200c이급성별결정구Y광단백-2(SOX-2)적표체.결과 내약주적내약성현저고우친본주(P＜0.01).As2O3강저료내약주대CDDP적내약성,역전배수위3.39,상대역전효솔위76.6％.상대우친본주,내약주세포let-7화miR-200c적표체현저강저(P＜0.05),SOX-2적표체현저승고(P＜0.01).As2 03처리후,내약주세포let-7화miR-200c적표체현저증가(P＜0.05),SOX-2적표체현저하강(P＜0.05).결론 CSC유가능시인골육류세포주MG63대CDDP산생내약적궤제지일.
Objective To investigate the role of cancer stem cells (CSC) in the acquired resistance to cisplatin (CDDP) of human osteosarcoma cell line MG63.Methods CDDP-resistant human osteosarcoma cell line MG63 (MG63/CDDP) was established in vitro by exposure to high and gradually increased dose of cisplatin (1,3,5 mg/L).The proliferation and sensitivity to CDDP of MG63/CDDP before and after application of As2O3 (0.20 mg/L) were tested using methyl thiazol tetrazolium (MTT) assay.Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to investigate the expression of let-7,microRNA (miR)-200c and sex determining region Y-box 2 (SOX-2) in MG63/CDDP cells before and after application of As2O3.Results (1) In the presence of CDDP (3 mg/L),the growth rate of MG63/CDDP was significantly higher compared with MG63 cells (P ＜0.01).It was also demonstrated that MG63/CDDP treated by CDDP developed similar proliferation to MG63 cells without treatment of CDDP (P ＞ 0.05).The IC50 of MG63 and MG63/CDDP cells to CDDP were about 6.3 mg/L and 78.9 mg/L respectively and the resistance index was about 12.52.(2) The resistance of MG63/CDDP cells to CDDP was reduced by treatment of As2O3 (0.20 mg/L) with reversed multiples of 3.39 and relative reversed efficiency of 76.6％.(3) MG63/CDDP cells expressed reduced level of let-7 and miR-200c (P ＜ 0.05) and elevated expression of SOX-2 (P ＜0.01) in comparison with MG63 cells.(4) After treatment of As2O3,MG63/CDDP cells developed increased expression of let-7 and miR-200c (P ＜ 0.05) and decreased level of SOX-2 (P ＜ 0.05).Conclusion CSC is a possible mechanism accounting for the acquired resistance of MG63/CDDP to CDDP.