摘要
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 and Objective. Caveolin-1 is a marker protein of caveolae which is related with oncogenesis as a signal transducting hinge. This study was to investigate the effect of Caveolin-1 on the growth and apoptosis of doxorubicin-resistant human breast carcinoma cell line Hs578T/Dox. Methods. Plasmids pCI-neo-caveolin-1 and pCI-neo-vector (control) were transfected into Hs578T/Dox cells, respectively. The expression of Caveolin-1 was detected by Western blot. Cell proliferation was detected by MTT assay. Cell cycle was detected by flow cytometry (FCM). Colony formation potential on soft agar was evaluated. Cell apoptosis was detected by FCM when cells were cultured for 48 h or cultured with staurosporine for 8 h. Results. Caveolin-1 was overexpressed in Hs578T/ Dox-cav-1 cells. The proliferation of Hs578T/Dox-cav-1 cells was obviously promoted when compared with that of Hs578T/Dox-vector cells (P〈0.01). Colony size was larger in Hs578T/Dox-cav-1 group than in Hs578T/Doxvector group. More colonies were formed in Hs578T/Dox-cav-1 group as compare with those in Hs578T/Dox-vector group (983.6±75.0 vs. 700,8± 78.9, P〈0.01). The proportions of cells at S and G2/M phases were higher in Hs578T/Dox-cav-1 group than in Hs578T/Dox-vector group. The proliferation rate of Hs578T/Dox-cav-1 cells was also higher than that of Hs578T/Doxvector cells E(76.6±4.0)% vs. (58.0±4.1)% ]. Over-expressed Caveolin-1 significantly reduced apoptosis index when the cells were cultured for 48 h [ (5.7±0.5)% vs. (11.3±0.8)%] or culuted with staurosporine for 8 h [ (13.8± 1.2)% vs (21.4±1.9)%]. Conclusion: Overexpression of Caveolin-1 protein may promote the growth and anti-apoptosis ability of Hs578T/Dox cells.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2009年第6期587-592,共6页
Chinese Journal of Cancer