靛玉红衍生物PHⅡ-7通过抑制c-fos表达抗乳腺癌耐药株MCF-7/ADR增殖

Indirubin derivative PHⅡ-7 suppresses the proliferation of resistant human breast cancer MCF-7/ADR cells via inhibiting c-fos expression

目的:探讨靛玉红衍生物PHⅡ-7抗乳腺癌耐药株MCF-7/ADR增殖作用的初步机制。方法:MTT法检测PHⅡ-7的体外杀伤活性;RT-PCR法、Western blot法检测基因、蛋白表达水平;构建干扰c-fos的短发夹RNA真核表达载体,转染MCF-7/ADR细胞,并建立稳定转染的细胞系;采用细胞计数法绘制生长曲线探讨c-fos表达下调对乳腺癌细胞增殖的影响。结果:PHⅡ-7剂量依赖性地抑制乳腺癌敏感株MCF-7及耐药株MCF-7/ADR增殖;c-fos在耐药株MCF-7/ADR表达明显高于敏感株MCF-7;PHⅡ-7明显抑制c-fos的表达;shRNA显著抑制了c-fos蛋白表达,细胞增殖也被显著抑制。结论:靛玉红衍生物PHⅡ-7浓度依赖性地抑制乳腺癌耐药株MCF-7/ADR增殖,其作用可能与抑制原癌基因c-fos有关。

Objective：To detect the primary mechanism of the inhibitory effects of indirubin derivative PHIⅡ-7 on the proliferation of resistant breast cancer MCF-7/ADR cells. Methods：Human breast cancer sensitive MCF-7 and multidrug resistant MCF-7/ADR cells were treated with dif-ferent concentrations of PH 11-7 ,then the proliferation of cells was examined using MTT assay ; RT-PCR and western blot analysis were used to investigate the variation of mRNA and protein levels,respectively;construct the short hairpin RNA targeting c-fas and develop the stable MCF- 7/ADR cell line expressing this shRNA; growth curve assay by counting cells was performed to explore the effect of c-fos inhibition on the pro- liferation of breast cancer cells. Results ： PH Ⅱ-7 showed its inhibitory effects on both sensitive MCF-7 and resistant MCF-7/ADR cells in a concentration dependent manner;elevated c-fos expression was detected in MCF-7/ADR compared with MCF-7 cells;PH Ⅱ-7 significantly in- hibited the expression of pro-oncogene c-fos ; shRNA remarkably inhibited the protein expression of c-fos as well as the proliferation of breast cancer cells （P 〈 0.05 ）. Conclusion：The pro-oncogene c-fos might be involved in the inhibitory effects of PH Ⅱ-7 on the proliferation of re- sistant human breast cancer MCF-7/ADR cells.