COX-2选择性抑制剂联合奥沙利铂对结肠癌HCT-8细胞增殖和凋亡的影响

Effects of COX-2 selective inhibitor combined with Oxaliplatin on cells' proliferation and apoptosis of HCT-8 cells of colorectal carcinoma

目的观察环氧合酶-2(COX-2)选择性抑制剂NS-398联合奥沙利铂(L-OHP)对结肠癌HCT-8细胞增殖和凋亡的影响,比较NS-398联合L-OHP与单独应用L-OHP对结肠癌HCT-8细胞的杀伤效应。方法 CCK-8法检测不同浓度的L-OHP(1.25、2.50、5.00、10.00及20.00μmol/L)单独作用及L-OHP联合NS-398(20.00及80.00μmol/L)作用HCT-8细胞24 h的细胞增殖抑制率。分别应用流式细胞术检测细胞早期凋亡率;Hoechst33258染色观察细胞凋亡形态学改变;分光光度法检测Caspase-3的活性。结果细胞增殖抑制实验表明L-OHP、NS-398对HCT-8细胞有剂量依赖性抑制增殖作用(P<0.05)。NS-398联合L-OHP用药组细胞增殖抑制率明显高于同浓度L-OHP单用组,且随着添加的NS-398剂量的增加而增加,联合用药组对HCT-8细胞的诱导凋亡效应明显高于L-OHP单药组,二者具有协同效应。两药均能提高Caspase-3的活性,联合用药组中Caspase-3的活性显著高于L-OHP组和NS-398组(P<0.01)。结论 NS-398和L-OHP均能抑制HCT-8细胞的增殖及诱导凋亡,NS-398能够显著增强L-OHP的癌细胞增殖抑制及诱导凋亡作用。

[Objective] To observe the role of cyclooxygenase-2 （COX-2） selective inhibitor NS-398 combined with Oxaliplatin （L-OHP） in human calorectal carcinoma HCT-8 cell line on proliferation and apoptosis. The antitu- mot effect of NS-398 combined with L-OHP and L-OHP alone on HCT-8 cells were compared. [ Methods ] The inhibition rate of HCT-8 cells treated by different concentration of L-OHP （1.25, 2.50, 5.00, 10.00 and 20.00μmol/L） alone and L-OHP combined with NS-398 （20 and 80 Ixmol/L） for 24 h was detected by CCK-8 kit. Flow cytometry, Hoechst33258 staining and spectrophotometric assay were also used to analyze early apoptotic rate, apoptotic mor- phological changes and Caspase-3 activity, respectively. [ Results ] Cell proliferation assay showed that L-OHP and NS-398 had a dose-dependent inhibition effects on HCT-8 cells, respectively （P 〈0.05）. The inhibition rate in NS- 398 combined with L-OHP group was higher than that of the same concentration of L-OHP group, and as the dosage of NS-398 increasing, the inhibition effects increased. The apoptosis inducing effects in combined treatment group on HCT-8 cells were significantly higher than that of L-OHP group, the two drugs had a synergistic effect. Both of NS-398 and L-OHP could increase easpase-3 activity. Caspase-3 activity in the combined treatment group was significantly higher than that of L-OHP group and/or NS-398 group （P 〈0.01）. [Conclusions] Both of NS-398 and L-OHP could inhibit HCT-8 cell proliferation and induce apoptosis. NS-398 could significantly enhance the effects of L-OHP on HCT-8 cells.