摘要
目的研究caveolin-1对乳腺癌细胞MCF7在体内、体外生长和增殖的影响并初步探讨其机制。方法选用人类乳腺癌细胞MCF7,通过基因转染技术培育过表达caveolin-1的细胞株MCF7/cav-1作为实验组,空载体细胞株MCF7/vec作为对照组。Western blot方法检测转染细胞内caveolin-1和血管内皮生长因子(VEGF)的表达。软琼脂集落形成实验检测细胞增殖克隆的能力。建立裸鼠皮下种植瘤模型并检测肿瘤组织中Ki-67与VEGF的表达情况。结果 caveolin-1在MCF7/cav-1细胞中表达稳定,其表达量明显高于对照组细胞MCF7/vec(P<0.01)和亲本细胞MCF7(P<0.01),而对照组细胞MCF7/vec和亲本细胞MCF7之间caveolin-1表达量差异无统计学意义(P>0.05)。与MCF7/vec细胞比较,MCF7/cav-1细胞在软琼脂中形成的集落数目明显减少(P<0.01),体积也较小。MCF7/cav-1细胞中VEGF表达量明显低于MCF7/vec细胞(P<0.01)。在裸鼠的体内实验中,与MCF7/vec细胞比较,MCF7/cav-1细胞在裸鼠体内生长缓慢(P<0.01),Ki-67染色明显减弱,VEGF染色阴性,提示caveolin-1可以抑制肿瘤体内增殖。结论 caveolin-1可能通过抑制VEGF表达抑制MCF7细胞生长和增殖。
Objective To investigate the effect of caveolion-1 on the growth and proliferation in human breast cancer MCF7 cell in vitro and in vivo and approach its mechanisms.Methods The plasmid pCI-neo-cav-1 and its corresponding empty vector（pCI-neo） were transfected into MCF7 cell line.The expressions of caveolin-1 and vascular endothelial growth factor（VEGF） in transfectants were subsequently confirmed by Western blot analysis.A pair of monoclonal cell lines,MCF7/cav-1 and MCF7/vec,were chose for future studies.The colony formation ability of tumor cells was detected by anchorage-independent growth assay.Xenograft tumor models in nude mice were established.Immunohistochemistry was used to characterize Ki-67 and VEGF levels in the tumors tissues.Results Caveolin-1 expression was up-regulated in the MCF7/cav-1 cell as compared with the MCF7/vec cell（P0.01） and the MCF7 cell（P0.01）.Caveolin-1 overexpression markedly reduced the capacity of the cells to form colonies in soft agar（P0.01）.Compared with the MCF7/vec cell,VEGF protein expression reduced in the MCF7/cav-1 cell（P0.01）.In vivo experiments in nude mice,the overexpression of caveolin-1 resulted in significant growth inhibition of the xenograft breast tumors.The Ki-67 staining was weak and the VEGF staining was negative by immunohistochemistry that indicated the caveolin-1 inhibited the proliferation in human breast cancer MCF7 cell.Conclusion The caveolin-1 might inhibit the growth and proliferation in human breast cancer MCF7 cell through suppressing activation of VEGF signaling pathway. ？？
出处
《中国普外基础与临床杂志》
CAS
2013年第5期481-486,共6页
Chinese Journal of Bases and Clinics In General Surgery
