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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 investigate the primary mechanism of proliferative effect of indirubin derivative PHⅡ-7 on breast cancer cell line MCF-7 / ADR. Methods: The cytotoxicity of PHⅡ-7 in vitro was detected by MTT assay. The expression of gene and protein was detected by RT-PCR and Western blot. The eukaryotic expression vector of short hairpin RNA interfering with c-fos was constructed and transfected into MCF-7 / ADR Cells, and establish stable transfected cell lines; cell counting method of growth curve to investigate the c-fos expression down-regulation on breast cancer cell proliferation. Results: PHⅡ-7 dose-dependently inhibited the proliferation of breast cancer MCF-7 and MCF-7 / ADR. The expression of c-fos in MCF-7 / ADR was significantly higher than that of MCF-7. PHⅡ-7 significantly inhibited the expression of c-fos; shRNA significantly inhibited c-fos protein expression, cell proliferation was also significantly inhibited. CONCLUSION: The indirubin derivative PHⅡ-7 inhibits the proliferation of MCF-7 / ADR cells in a concentration-dependent manner, which may be related to the inhibition of proto-oncogene c-fos.