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背景与目的:作为信号转导枢纽的Caveolae与恶性肿瘤发生和耐药密切相关,Caveolin-1为其标志蛋白。本研究探讨Caveolin-1对肿瘤耐药细胞体外生长凋亡的影响及其作用机制。方法:选用乳腺癌Hs578T阿霉素耐药细胞株Hs578T/Dox,通过基因转染技术培育Caveolin-1蛋白过表达细胞株Hs578T/Dox-cav-1作为实验组,空载体转染细胞株Hs578T/Dox-vector作为对照组,Western blot检测细胞Caveolin-1蛋白表达。研究细胞生长凋亡变化:MTT检测细胞生长曲线,流式细胞技术细胞周期分析,检测软琼脂集落形成能力。细胞培养48h,流式细胞技术检测细胞自然凋亡指数;加入凋亡诱导剂-星形孢菌素细胞培养8h,检测星形孢菌素诱导细胞凋亡指数。结果:Hs578T/Dox-cav-1较Hs578T/Dox-vector的Caveolin-1蛋白稳定过表达。Hs578T/Dox-cav-1与Hs578T/Dox-vector细胞生长曲线比较,生长明显增快(P<0.01),软琼脂培养集落形成体积增大、数目增加[(983.6±75.0)vs(700.8±78.9),P<0.01]。细胞周期分析,Hs578T/Dox-cav-1细胞S期和G2/M期细胞比率增大,增殖指数升高[(76.6±4.0)%vs.(58.0±4.1)%]。细胞自然凋亡指数[(5.7±0.5)%vs.(11.3±0.8)%]和星形孢菌素诱导凋亡指数[(13.8±1.2)%vs.(21.4±1.89)%]下降。结论:高表达Caveolin-1蛋白使乳腺癌阿霉素耐药细胞Hs578T/Dox具有更强的生长能力和抗凋亡能力。
BACKGROUND & OBJECTIVE: Caveolae, a signal transduction junction, is closely related to the occurrence and resistance of malignant tumors. Caveolin-1 is a marker protein. This study was to investigate the effect of Caveolin-1 on the growth and apoptosis of drug-resistant cells in vitro and its mechanism. Methods: Hs578T / Dox gene was transfected into Hs578T / Dox cell line Hs578T / Dox which was an overexpressed cell line of H578T / Dox-cav-1, Dox-vector as a control group, Western blot detection of the expression of Caveolin-1 protein. The apoptosis of cell growth was studied: MTT assay cell growth curve, flow cytometry cell cycle analysis, detection of soft agar colony formation ability. The cells were cultured for 48h and the apoptosis index was detected by flow cytometry. The apoptotic index induced by staurosporine was detected by adding the apoptosis inducer-staurosporine for 8h. Results: Hs578T / Dox-cav-1 stably overexpressed Caveolin-1 protein of Hs578T / Dox-vector. Compared with Hs578T / Dox-vector cell growth curve, the growth of Hs578T / Dox-cav-1 was significantly increased (P <0.01) ), P <0.01]. Cell cycle analysis showed that the percentage of cells in S phase and G2 / M phase of Hs578T / Dox-cav-1 cells increased and the proliferation index increased (76.6 ± 4.0) vs (58.0 ± 4.1)%. The apoptotic index [(5.7 ± 0.5)% vs (11.3 ± 0.8)%] and staurosporine induced apoptosis index [(13.8 ± 1.2)% vs (21.4 ± 1.89)%] decreased. CONCLUSION: High expression of Caveolin-1 protein enhances the ability of adriamycin-resistant cells Hs578T / Dox to grow and anti-apoptotic.