芬太尼对MCF-7细胞增殖和细胞周期影响研究

The Effect of Fentanyl on Proliferation and Cell Cycle of Human Breast Carcinoma Cell Line MCF-7

目的 探讨芬太尼对MCF -7细胞增殖和细胞周期的影响及其机制。方法 MCF -7细胞在不同浓度芬太尼、纳洛酮和两药联合干预后 ,采用MTT法检测细胞增殖活性 ;流式细胞术检测细胞周期 ;免疫细胞化学染色SP法检测p5 3、p2 1/WAF1的表达。结果 芬太尼浓度≥ 0 1μmol/L时 ,细胞增殖活性较对照组显著下降 (P <0 0 5 ) ,其效应与时间和浓度均呈正相关 (P <0 0 1) ,72h半数抑制浓度 (IC50 )为 0 81± 0 0 2 μmol/L。随着芬太尼浓度增加 ,G0 /G1期比例逐渐增加 ,(S +G2 /M )期比例逐渐减少。芬太尼组浓度≥ 0 1μmol/L时 ,细胞增殖活性较对照组显著下降 (P <0 0 5 ) ,较纳洛酮和芬太尼联合给药组差异无显著性。给予芬太尼 10 μmol/L后胞核内p5 3、p2 1/WAF1AOD显著增加 (P <0 0 5 )。结论 芬太尼浓度和时间依赖性抑制MCF -7细胞增殖 ,主要是将细胞阻滞在G1期 ;其机制可能是上调野生型p5 3和p2 1/WAF1的表达。

Objective To study the effects and its mechanism of fentanyl(Fen) on the proliferation and cell cycle of human breast carcinoma line MCF-7. Methods MCF-7 cells were cultured in the medium with Fen, naloxone(Nx) or both the medicines at different concentration for different time. MTT method was employed to evaluate the level of the cell proliferation. The distribution of the cell cycle was detected with the flow cytometry (FCM). The expression levels of p53 and p21/WAF1 in the cells were determined by SP immunocytochemical staining method. Results Fen at≥0.1μmol/L concentration inhibited MCF-7 cells proliferation in dose- and time-dependent manners, and its IC 50 for 72h was 0.81±0.02 μmol/L. However, the antiprolifeative effect of Fen was not antagonized by Nx. Fen significantly enhanced the ratio of G 0/G 1 phase MCF-7 cells, and decreased the proliferation index of MCF-7 cells in dose-dependent manner. Fen also upregulated the expression of p53 and p21/WAF1 in MCF-7 cells. Conclusion The data suggested that the inhibitory effect of Fen on MCF-7 cell growth might be mediated by blocking cell cycle progression from G 1 to S phase, and upregulating the expression of p53 and p21/WAF1.