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目的观察5氮2脱氧胞苷(5-AZA-CDR)对雌激素受体Α(ERΑ)阴性人类乳腺癌细胞株MDA-MB-231和MDA-MB-435ERΑ基因诱导表达作用;5-AZA-CDR联合三苯氧胺(TAM)对ERΑ阴性乳腺癌细胞的体外抑制作用。方法应用甲基化特异性PCR(MSP)检测MDA-MB-231和MDA-MB-435细胞和20例ERΑ阴性乳腺癌组织ERΑ基因核心启动子区CPG岛甲基化情况;5-AZA-CDR处理MDA-MB-231和MDA-MB-435细胞,逆转录PCR(RT-PCR)检测ERΑMRNA表达;5-AZA-CDR、TAM分别或联合作用于MDA-MB-231和MDA-MB-435细胞,MTT比色法分析细胞生长抑制作用,流式细胞仪测定细胞周期分布和凋亡率。结果适当浓度的5-AZA-CDR能诱导MDA-MB-231和MDA-MB-435细胞表达ERΑMRNA,抑制细胞生长、阻滞细胞周期于G0/G1期,诱导细胞凋亡;TAM对MDA-MB-231和MDA-MB-435细胞生长、细胞周期无影响;而两药联合时能显著抑制细胞生长,诱导细胞凋亡,凋亡率分别为48.8%和53.1%。结论5-AZA-CDR能诱导ERΑ阴性乳腺癌表达ERΑMRNA,恢复ERΑ阴性乳腺癌细胞对TAM的敏感性,联合TAM能协同抑制ERΑ阴性乳腺癌细胞生长,诱导细胞凋亡,从而为ERΑ阴性乳腺癌开辟新的内分泌治疗途径提供实验依据。
Objective To observe the effect of 5-AZA-CDR on the expression of MDA-MB-231 and MDA-MB-435ERA in human esophageal cancer cell line ERA-negative human breast cancer cells. Inhibitory effect of CDR combined with tamoxifen on ERα negative breast cancer cells in. Methods Methylation-specific PCR (MSP) was used to detect the methylation of CPG island in the core promoter region of ERA gene in MDA-MB-231 and MDA-MB-435 cells and in 20 ERA negative breast cancer tissues. The 5-AZA- The MDA-MB-231 and MDA-MB-435 cells were treated with 5-AZA-CDR and TAM, respectively. The expression of ERA mRNA was detected by reverse transcription polymerase chain reaction (RT- MTT colorimetry was used to analyze the cell growth inhibition. Flow cytometry was used to determine the cell cycle distribution and apoptosis rate. Results The 5-AZA-CDR at the appropriate concentration could induce the expression of ERAMM in MDA-MB-231 and MDA-MB-435 cells and inhibit the cell growth and arrest the cell cycle at G0 / -231 and MDA-MB-435 cells. The combination of the two drugs could significantly inhibit cell growth and induce apoptosis, with the apoptosis rates of 48.8% and 53.1%, respectively. Conclusions 5-AZA-CDR can induce the expression of ERA mRNA in ER-negative breast cancer and restore the sensitivity of ER-negative breast cancer cells to TAM. Combining with TAM can inhibit the growth of ER-negative breast cancer cells and induce apoptosis, Open up new ways of endocrine therapy to provide experimental evidence.