4-氨基吡啶对多西紫杉醇抗人乳腺癌细胞作用影响的实验研究

Experimental studies on the effects of 4-AP on the action of docetaxel inhibits human breast cancer cell

目的:探讨选择性钾离子通道阻断剂4-氨基吡啶(4-AP)对多西紫杉醇(do-cetaxel,DOC)抗人乳腺癌细胞MDA-MB-43-5S增殖、凋亡和周期的影响。方法:MTT比色法分析DOC、4-AP单独应用以及两药联合应用对MDA-MB-435S增殖的影响;流式细胞术Annexin V-FITC/PI双染法和PI单染法检测两种药物对MDA-MB-435S凋亡和周期的影响。结果:DOC和4-AP单独应用均能抑制MDA-MB-435S的增殖;5mmol/L的4-AP并用25μmol/L的DOC对MDA-M-B-435S达最大抑制率的时间明显缩短,DOC和4-AP对MDA-MB-435S的增殖抑制有相加或协同作用,并呈剂量-时间依赖性。2和5μmol/L的DOC单独作用24h,MDA-M-B-435S产生凋亡并不明显,当与5mmol/L的4-AP联合应用后,细胞早期凋亡和死亡明显增加。单用DOC可使MDA-MB-435S阻断在G2/M期,单用4-AP可使MDA-M-B-435S阻断在G0/G1期,两药联合应用可使MDA-MB-435S阻断在G2/M期。结论:DOC和4-AP分别在G2/M期和G0/G1期抑制MDA-MB-435S的增殖,4-AP通过促进DOC诱导细胞凋亡从而抑制MDA-MB-43-5S的增殖。

OBJECTIVE： To study the effects of selective K^＋ channel blocker 4-AP on the action of DOC inhibits human breast cancer cell MDA-MB-435S on the cell proliferation, apoptosis and cell cycle. METHODS： The effects of DOC, 4-AP and the combination of both drugs on proliferation of MDA-MB-435S cell were investigated by MTT assays. The apoptosis cells and cell cycles were measured by flow cytometry after the cells were stained by both Annexin-V and PI and PI only, respectively. RESULTS： DOC and 4-AP inhibited the proliferation of human breast cancer cell MDA-MB-435S, respectively. 4-AP of 5 mmol/L advanced the time of DOC of 25 μmol/L inducing human breast cancer cell MDA-MB-435S to the maximum inhibitory rate. DOC and 4-AP had additive or synergetic effects on inhibiting the proliferation of MDA-MB-435S, and the effects presented the time-dose dependence. The effects of improving MDA-MB-435S cell apoptosis were not obvious when given DOC of 2 μmol/L or 5 μmol/L for 24 hours, but when DOC were combined with 4-AP of 5 mmol/L, both the pristine cell apoptosis and the cell death were very transparent. MDA-MB-435S cells were blocked at G2/M period by DOC, they were blocked at G0/G1 period by 4-AP, and they were blocked at G2/M period when using both DOC and 4-AP. CONCLUSIONS： DOC and 4-AP can inhibit human breast cancer cell MDA-MB-435S proliferation by blocking cells at G2/M period and G0/G1 period, respectively. 4-AP can inhibit MDA-MB-435S proliferation by improving cell apoptosis induced by DOC.