CAJ | 학술논문

目的：通过顺铂（cisplatin，DDP）诱导卵巢癌细胞珠 SKOV3建立耐药细胞株 SKOV3/DDP，并研究其与凋亡途径蛋白的关系。方法应用倒置显微镜观察 DDP 对细胞形态的影响，四甲基偶氮唑蓝（methyl thiazolyl tetrazolium，MTT）法检测 DDP 对 SKOV3和 SKOV3/DDP 细胞的增殖抑制情况，流式细胞术（flow cytometry， FCM）检测细胞凋亡率以及细胞中 X链锁凋亡抑制蛋白（X-linked inhibitor-of-apoptosis protein，XIAP）、半胱氨酸天冬氨酸蛋白酶3（cysteine-aspartic acid protease-3，Caspase-3）、B 细胞淋巴瘤/白血病2（B cell lymphoma/lewkmia-2，Bcl-2）和生存素（Survivin）的表达情况。结果 MTT法检测发现DDP作用后，SKOV3/DDP细胞的增殖抑制率较 SKOV3细胞显著降低（P＜0．01），且存在着浓度和时间依赖效应（P＜0．01）。由半数抑制浓度（50％concentration of inhibition，IC50）比较得 SKOV3/DDP 细胞24、48、72h 耐药指数（resistance index，RI）分别为2．434、2．950、3．780。FCM检测表明，DDP 作用后，SKOV3/DDP 凋亡率显著低于 SKOV3细胞（P＜0．01）。与SKOV3细胞相比，SKOV3/DDP细胞中XIAP、Bcl-2、Survivin蛋白高表达，Caspase-3蛋白低表达，差异有统计学意义（P＜0．01）。结论成功建立卵巢癌耐药细胞株 SKOV3/DDP，其耐药机制可能与 XIAP、Caspase-3、Bcl-2和Survivin蛋白的异常表达有关，表明这些蛋白参与了卵巢癌DDP耐药的形成。
목적：통과순박（cisplatin，DDP）유도란소암세포주 SKOV3건립내약세포주 SKOV3/DDP，병연구기여조망도경단백적관계。방법응용도치현미경관찰 DDP 대세포형태적영향，사갑기우담서람（methyl thiazolyl tetrazolium，MTT）법검측 DDP 대 SKOV3화 SKOV3/DDP 세포적증식억제정황，류식세포술（flow cytometry， FCM）검측세포조망솔이급세포중 X련쇄조망억제단백（X-linked inhibitor-of-apoptosis protein，XIAP）、반광안산천동안산단백매3（cysteine-aspartic acid protease-3，Caspase-3）、B 세포림파류/백혈병2（B cell lymphoma/lewkmia-2，Bcl-2）화생존소（Survivin）적표체정황。결과 MTT법검측발현DDP작용후，SKOV3/DDP세포적증식억제솔교 SKOV3세포현저강저（P＜0．01），차존재착농도화시간의뢰효응（P＜0．01）。유반수억제농도（50％concentration of inhibition，IC50）비교득 SKOV3/DDP 세포24、48、72h 내약지수（resistance index，RI）분별위2．434、2．950、3．780。FCM검측표명，DDP 작용후，SKOV3/DDP 조망솔현저저우 SKOV3세포（P＜0．01）。여SKOV3세포상비，SKOV3/DDP세포중XIAP、Bcl-2、Survivin단백고표체，Caspase-3단백저표체，차이유통계학의의（P＜0．01）。결론성공건립란소암내약세포주 SKOV3/DDP，기내약궤제가능여 XIAP、Caspase-3、Bcl-2화Survivin단백적이상표체유관，표명저사단백삼여료란소암DDP내약적형성。
Objective To establish drug-resistant cell line SKOV3/DDP through ovarian cancer cell line SKOV3 by induced cisplatin (DDP ),and to study its relationship with the apoptosis pathway proteins.Methods The inverted microscope was applied to observe DDP influence on the cell morphology.Methyl thiazolyl tetrazolium (MTT)assay was used to detect proliferation inhibition rate affected by DDP on SKOV3 and SKOV3/DDP cells.Flow cytometry (FCM)was used to detect the influence of cell apoptosis and the expression of X-linked inhibitor-of-apoptosis protein (XIAP ),cysteine-aspartic acid protease-3 (Caspase-3 ),B cell lymphoma/lewkmia-2(Bcl-2)and Survivin proteins.Results MTT assay showed that proliferation inhibition rate of SKOV3/DDP cells decreased significantly compared with SKOV3 cells after DDP influence (P<0.01),and depended on the concentration and time (P<0.01).The resistance index of DDP to SKOV3/DDP and SKOV3 cells in 24h,48h and 72h were respectively 2.434,2.950 and 3.780 by comparison between 50% concentration of inhibition values.FCM revealed that the apoptosis rates of SKOV3/DDP cells to DDP decreased obviously after 48h than those of SKOV3 cells (P<0.01).Compared with SKOV3 cells,the expression of XIAP,Bcl-2 and Survivin proteins in SKOV3/DDP cells were higher,while the expression of Caspase-3 protein was lower,and their differences were statistically significant (P<0.01).Conclusion Drug-resistant cell line SKOV3/DDP for ovarian cancer is successfully established and its resistance mechanism is closely related with the abnormal expression of XIAP,Caspase-3,Bcl-2 and Survivin proteins.These proteins might be involved in the drug resistance of ovarian cancer.