顺铂对上皮性卵巢癌细胞SKOV-3中Ca^(2+)浓度的影响及其与化疗耐药的关系研究

Effect of Cisplatin on Intracellular-free Calcium Level of SKOV-3 and it's Relationship with the Acquired Drug Resistance

目的探讨顺铂对上皮性卵巢癌细胞SKOV-3中Ca2+浓度的影响,并分析Ca2+浓度的变化与细胞获得性耐药的关系。方法体外培养SKOV-3细胞(对照组)以及SKOV-3顺铂耐药细胞株(实验组),并通过MTT法分析顺铂对两种细胞株的抑制作用;采用Fura2-AM标记细胞内Ca2+,通过荧光分析仪测定实验组与对照组SKOV-3细胞内Ca2+浓度。通过western-blot检测比较两组细胞E-cadherin、Vimentin表达水平的差异。通过粘附实验、划痕实验以及Transwell法比较两组细胞的粘附、迁移和侵袭能力。结果实验组细胞与对照组细胞对顺铂的IC50值分别为83.27μg·mL-1以及18.32μg·mL-1,实验组细胞内Ca2+浓度为(187.31±11.37)nmo·lL-1,明显高于对照组细胞内的Ca2+浓度(135.71±15.52)nmo·lL-1,差异有统计学意义(P<0.05)。实验组细胞E-cadherin与Vimentin蛋白表达水平明显高于对照组,两组蛋白OD值差异有统计学差异(P<0.05)。实验组细胞粘附、迁移以及侵袭能力明显强于对照组细胞,相关指标差异有统计学意义。结论耐顺铂的SKOV-3细胞内Ca2+浓度升高,该离子浓度水平与细胞的粘附、迁移以及侵袭能力存在关联性。

Objective To explore the effect of cisplatin on the intracellular-free calcium level of SKOV-3 and it＇ s relation- ship with the acquired resistance to the cisplatin. Method The SKOV-3 （control） and cisplatin-resistant SKOV-3 （experiment group） were cultured in vitro. The MTT method was used to analyze the cisplatin sensitivity of experiment group and control group. The Ca2＋ was marked with Fura2-AM, and its level was detected by spectrofluorimeter. The expression levels of E-cad- herin and Vimentin in two groups were tested by Western-blot. The adhesion test, wound healing assay and Transwell invasion assay were used to test the cell adhesion, migration and invasion ability of the two groups. Results The IC50 of the experiment group and control group were respectively 83.27 μg.mL^-1 and 18.32 μg.mL^-1. The difference was significant （P〈0.05）. The Ca2＋ level in the experiment group was （187.31 ± 11.37） nmol.L-1, significandy higher than that of the control group which was （135.71 ± 15,52） nmol.L^-1, with statistical difference （P〈 0.05）. The expression levels of E-cadherin and Vimentin of the experiment group were significantly higher than those of control group, and the protein OD value showed statistical difference （P〈 0.05）. The adhesion ability, migration distance and the number of invasive cells of the experiment group were significantly higher or more than those of the control group （P 〈 0.05）. Conclusion The Ca2＋ level of the cisplatin-resistant SKOV-3 cell was significantly in- creased, and has strong relationship with the cell adhesion, migration and invasion ability.