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目的探索利用TGF-β1诱导乳腺癌细胞MCF-7 EMT水平,并进行生物学验证。方法培养人乳腺癌细胞系MCF-7并传代,使用不同浓度的TGF-β1因子进行诱导,用MTT法检测TGF-β1对乳腺癌细胞的生长促进作用,用实时荧光定量PCR技术对上皮细胞-间质转化的标志物Vimentin表达水平进行检测,利用Transwell小室迁移试验对诱导后的迁移能力进行评估和检测。结果浓度为12.5 ng/ml的TGF-β1诱导MCF-7细胞48 h后细胞形态呈现EMT改变,提示有上皮细胞间质化发生,MTT检测结果显示TGF-β1对乳腺癌细胞MCF-7生长增殖无显著影响,对照组与实验组之间比较,差异无统计学意义(P>0.05);EMT间质细胞标志物Vimentin表达上调;Transwell小室迁移试验结果显示,对照组与实验组之间比较,差异有统计学意义(P<0.05)。结论人乳腺癌细胞系MCF-7经浓度为12.5 ng/ml的TGF-β1因子诱导48 h后呈现EMT改变,间质化标志物Vimentin表达上调,侵袭转移能力增强。
Objective To explore the use of TGF-β1-induced breast cancer cell MCF-7 EMT levels and biological verification. Methods Human breast cancer cell line MCF-7 was cultured and passaged, and induced by different concentrations of TGF-β1. MTT assay was used to detect the growth promoting effect of TGF-β1 on breast cancer cells. Real-time fluorescence quantitative polymerase chain reaction The expression of Vimentin, a marker of interstitial transformation, was measured and the posttranslational migration ability was evaluated and tested using Transwell chamber migration assay. Results The morphology of MCF-7 cells induced by TGF-β1 at a concentration of 12.5 ng / ml for 48 h showed EMT changes, suggesting epithelial cell interstitialization. The results of MTT assay showed that TGF-β1 promoted the proliferation of MCF-7 breast cancer cells (P> 0.05). The expression of EMT stromal cell marker Vimentin was up-regulated. The results of Transwell migration assay showed that there was no significant difference between the control group and the experimental group (P> 0.05) The difference was statistically significant (P <0.05). Conclusion The human breast cancer cell line MCF-7 is induced by TGF-β1 at a concentration of 12.5 ng / ml for 48 h. The EMT changes and the expression of interstitial markers Vimentin are up-regulated. The invasion and metastasis of MCF-7 is enhanced.