基于上皮间充质转化研究复方守宫散抑制三阴性乳腺癌恶性表型作用及机制研究

Effect and mechanism of Compound Shougong Powder in attenuating triple-negative breast cancer progression by reversing epithelial-to-mesenchymal transition

研究复方守宫散(Compound Shougong Powder,CSGP)对三阴性乳腺癌细胞生物学功能的作用及其作用机制是否与上皮间充质转化信号通路有关。三阴性乳腺癌细胞(MDA-MB-231和BT-549)分别给予不同浓度的复方守宫散含药血清,细胞增殖检测试剂盒(MTS)法检测复方守宫散对三阴性乳腺癌细胞增殖的影响;EdU染色检测复方守宫散对三阴性乳腺癌细胞增殖的影响;采用流式细胞术检测复方守宫散对三阴性乳腺癌细胞凋亡的影响;采用划痕实验和Transwell实验检测不同浓度的复方守宫散对三阴性乳腺癌细胞迁移、侵袭能力的影响;采用RNA测序技术阐明其机制;采用qRT-PCR技术检测E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、锌指转录因子(Slug)、蜗牛家族转录抑制因子(Snail)、波形蛋白(Vimentin)、扭曲基因(Twist)、E盒结合锌指蛋白1(Zinc finger E-box-Binding homeobox 1,Zeb1)、E盒结合锌指蛋白2(Zinc finger E-box-bin-ding homeobox 2,Zeb2)mRNA表达水平;采用Western blot检测Slug、Vimentin和E-cadherin蛋白表达水平。复方守宫散干预后,MDA-MB-231、BT-549细胞增殖明显减少,凋亡率显著升高,上皮标志蛋白E-cadherin表达水平显著升高,而上皮间充质转化相关转录因子Slug及Vimentin表达水平显示降低。综上所述,复方守宫散通过抑制上皮间充质转化,进而抑制三阴性乳腺癌恶性进展。

The study investigated the effect of Compound Shougong Powder(CSGP)on the biological functions of triple-negative breast cancer(TNBC)cells and whether its mechanism of action was related to the epithelial-mesenchymal transition(EMT)signaling pathway.TNBC cells(MDA-MB-231 and BT-549)were treated with different concentrations of CSGP-containing serum.MTS assay was used to detect the effect of CSGP on the proliferation of TNBC cells.The EdU staining was used to detect the effect of CSGP on the proliferation of TNBC cells.Flow cytometry was used to examine the impact of CSGP on apoptosis of TNBC cells.Wound-healing and Transwell assays were used to evaluate the effects of different concentrations of CSGP on the migration and invasion capabilities of TNBC cells.RNA sequencing technology was utilized to elucidate its mechanism.Subsequently,qRT-PCR was performed to measure the mRNA expression levels of E-cadherin,N-cadherin,Slug,Snail,Vimentin,Twist,Zinc finger E-box-Binding homeobox 1(Zeb1),and Zinc finger E-box-Binding homeobox 2(Zeb2).Western blot was used to assess the protein expression levels of Slug,Vimentin,and E-cadherin.After intervention with CSGP,the proliferation of MDA-MB-231 and BT-549 cells significantly decreased,while the apoptosis rate markedly increased.The expression levels of the epithelial marker protein E-cadherin significantly increased,while the expression levels of the EMT-related transcription factors Slug and Vimentin showed a decrease.In conclusion,CSGP inhibits the EMT,thereby suppressing the malignant progression of TNBC.