硼替佐米对人卵巢癌耐顺铂细胞株的生长抑制作用

Experimental study on the reversal effect of bortezomib on cisplatin-resistant human ovarian cancer cell lines

目的探讨硼替佐米对人卵巢癌耐顺铂细胞株（SKOV3／DDP）的逆转作用及其可能机制。方法体外培养人卵巢癌细胞株（SKOV3）和SKOV3／DDP，空白对照组加RPMI-1640完全培养基；硼替佐米组浓度分别为0．0625、0．1250、0．2500、0．5000、1．0000、2．0000μmol／L，检测不同作用时间对SKOV3／DDP的生长抑制情况；采用流式细胞术检测0．5000μmol／L硼替佐米作用后细胞周期及凋亡率的变化。结果①硼替佐米对SKOV3／DDP细胞生长抑制情况：0．0625、0．1250、0．2500、0．5000、1．0000、2．0000μmol／L的硼替佐米作用SKOV3／DDP细胞24h的生长抑制率依次为（1．19±0．07）％、（2．24±0．08）％、（3．47±0．20）％、（4．61±0．07）％、（5．80±0．17）％、（6．43±0．10）％；作用48h的生长抑制率依次为（9．39±0．08）％、（12．17±0．23）％、（18．08±0．25）％、（41．11±0．10）％、（55．45±0．41）％、（64．91±0．18）％；作用72h的生长抑制率依次为（13．21±0．32）％、（20．18±0．23）％、（22．91±0．35）％、（52．08±0．10）％、（76．59±0．39）％、（83．23±0．38）％。空白对照组抑制率为0，各实验组与空白对照组之间两两比较，差异均有统计学意义（均P〈0．05）。②细胞周期及凋亡率的变化情况：0．5000μmol／L硼替佐米作用SKOV3／DDP细胞48h，细胞周期G2／M期的比例为22．8％，空白对照组为10．1％，差异有统计学意义（P〈0．05）；凋亡率分别为11．7％和2．2％，差异有统计学意义（P〈0．05）。结论硼替佐米能够逆转SKOV3／DDP细胞的耐药作用；并将SKOV3／DDP的细胞周期阻滞于Gz／M期。

Objective To investigate the possible mechanisms that how bortezomib reverses the drug resistance of cisplatin-resistant human ovarian cancer cell lines （SKOV3/DDP）. Methods The resistant cell line-SKOV3/DDP and cell line-SKOV3 were chosen in vitro culture with different final density of bortezomib （0. 0625, 0. 1250, 0. 2500, 0. 5000, 1.0000, 2. 0000 μmol/L）. Different density and different action time of bortezomib were measured to investigate the impact on the growth suppression of SKOV3/DDP cell. The impact of 0. 5000 μmol/L bortezomib on the cell cycle and cell apoptosis of SKOV3/DDP and SKOV3 cell was measured by flow eytometry assay. Results ①After SKOV3/DDP cell in vitro culture with 0. 0625, 0. 1250, 0.2500, 0.5000, 1. 0000, 2. 0000 μmol/L bortezomib for 24 h, the rate of SKOV3/DDP cell growth suppression was （1. 19 ± 0.07）% ,（2.24 ±0.08）% ,（3.47 ±0.20）% ,（4.61 ±0.07）% ,（5.80 ±0.17% ） ,（6.43 ±0.10）% respectively;（9.39-±0.08）% ,（12.17 ±0.23）%, （18.08 ±0.25）% ,（41.11 ±0. 10）%, （55.45 ±0.41）% ,（64.91 ± 0.18）% after acting for 48 h; （13.21 ±0.32）%, （20. 18 ±0.23）%, （22.91 ± 0.35）%, （52.08 ±0.10）%, （ 76.59 ± 0.39 ） % , （ 83.23 ± 0.38 ） % after acting for 72 h. The different rate had statistical significant between experimental group and control group （ P 〈 0.05 ）. ②After 0. 5000 μmol/L bortezomib acting with SKOV3/DDP cells for 48 h, cell cycle GE/M phase ratio was 22.8% ;the control group was 10.1% and the difference had statistically significant （ P 〈 0.05 ） ; apoptosis rates were 11.7% and 2.2% ; the difference was statistically significant （ P 〈 0.05）. Conclusions Bortezomib can reverse the drug resistance of SKOV3/DDP. Bortezomib can block the cell cycle of cell line-SKOV3/DDP at the stage of G2/M.