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为了探索乳腺癌相关的人成纤维细胞中窖蛋白-1(Caveolin-1,Cav-1)与自噬体的相关性及其对乳腺癌细胞的作用,该研究采用si RNA技术干扰成纤维细胞株ESF表达Cav-1,q RTPCR和Western blot确定si RNA干扰Cav-1表达的效果;Transwell insert方法共培养乳腺癌细胞株BT474和ESF细胞,单丹磺酰戊二胺(monodansylcadaverin,MDC)染色、激光共聚焦显微镜观察Cav-1 si RNA对自噬体表达的影响;q RT-PCR和Western blot检测Cav-1 si RNA对微管相关蛋白1轻链3II(microtubule-associated protein 1 light chain 3 II,LC3II)表达的影响;CCK-8方法检测BT474细胞的增殖和活力。结果显示,靶向Cav-1的si RNA下调了ESF细胞中Cav-1的表达;Cav-1 si RNA促进ESF细胞自噬体和LC3II的表达,转染了Cav-1 si RNA的ESF细胞与BT474细胞共培养对自噬体和LC3II的作用更为显著;BT474细胞在ESFsi Cav-1(ESF cells transfected with Cav-1 si RNA)细胞共培养条件下增殖显著加快。研究表明,Cav-1 si RNA促进了与乳腺癌细胞共培养的成纤维细胞自噬体和LC3II的表达,同时加快了与成纤维细胞共培养的乳腺癌细胞的增殖。
In order to explore the relationship between Caveolin-1 (Cav-1) and autophagosome and its effect on breast cancer cells in breast cancer-related human fibroblasts, si RNA technology was used to interfere with the fibroblast The effect of si RNA on Cav-1 expression was determined by ESR expression of Cav-1, qRTPCR and Western blot. Transwell insert method was used to co-culture breast cancer cell lines BT474 and ESF cells. Monodansylcadaverin (MDC) staining , And the effect of Cav-1 si RNA on the expression of autophagosome was observed by laser scanning confocal microscopy. The expression of Cav-1 si RNA in microtubule-associated protein 1 light chain 3 (RT-PCR) II, LC3II) expression; CCK-8 method was used to detect the proliferation and viability of BT474 cells. The results showed that Cav-1 siRNA targeting Cav-1 downregulated the expression of Cav-1 in ESF cells; Cav-1 si RNA promoted the expression of autophagosomes and LC3II in ESF cells; and Cav-1 si RNA transfected ESF cells and BT474 cells co-cultured with autophagosomes and LC3II more significant role; BT474 cells in ESFsi Cav-1 (ESF cells transfected with Cav-1 si RNA) cells co-cultured proliferation significantly faster. Studies have shown that Cav-1 si RNA promotes fibroblast autophagosomes and LC3II expression in co-culture with breast cancer cells while accelerating the proliferation of breast cancer cells co-cultured with fibroblasts.