上皮性钙通道TRPV6对乳腺分化发育的影响  

The Effect of Epithelial Ca^(2+) Channel TRPV6 on Mammary Gland Differentiation

作　　者：韩学莹 赵丹 王建鹏 张玺 黄丽莉 李梦怡 董金堂[1] 赵强[1] Han Xueying;Zhao Dan;Wang Jianpeng;Zhang Xi;Huang Lili;Li Mengyi;Dong Jintang;Zhao Qiang(College of Life Sciences, Nanteai University, Tianjin 300071, China)

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2017年第5期76-83,共8页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：111基地经费支持(B08011);药物化学生物学国家重点实验室(南开大学)开放基金

摘　　要：为了研究上皮性钙通道TRPV6(transient receptor potential vanilloid 6)对乳腺分化发育的影响,本实验首先收集了发育不同时期的小鼠乳腺样本,共11个时间点,包括青春前期、青春期、孕期、泌乳期以及退化期,通过免疫荧光染色的方法观察了TRPV6在乳腺发育过程中的表达变化.其次,利用3维培养技术、Real-time PCR技术以及免疫荧光技术探究了TRPV6对正常乳腺上皮细胞MCF10A分化的影响.结果表明,在乳腺发育的不同阶段,TRPV6的表达水平具有明显差异,且在泌乳期达到最高;敲减TRPV6可以显著抑制MCF10A乳腺微球的形成,表现在成球数量的减少和微球平均直径的下降;在RNA和蛋白水平的检测结果显示TRPV6的敲减影响多个细胞分化相关分子的表达.上述结果提示TRPV6在乳腺发育过程中可能发挥重要作用.This study aims to investigate the effect of the calcium channel TRPV6 on mammary epithelial cell differentiation. In the beginning, we collected mouse mammary gland samples at different time points covering different developmental stages, which are postnatal, puberty, pregnancy, lactation, and involution, and then detected the expression of Trpv6 via immunofluorescence staining. We found expression of Trpv6 varied at developmental stages and peaked at lactation, indicating its potential role in luminal epithelial cell differentiation. Then, we explored the effect of TRPV6 using the in vitro 3-D culture model of the normal human mammary epithelial cell line MCF10 A. We observed that knockdown of TRPV6 interrupted mammosphere formation in the 3-D culture of MCF10 A cells, with both a dramatic decline in the number of mammospheres formed and also a decrease of average sphere size. By doing real-time PCR and immunofluorescence staining, we also found the expression of multiple cell differentiation related molecules was altered after TRPV6 knockdown. These finding indicate that TRPV6 plays a role in the development of mammary gland likely.

关 键 词：乳腺 细胞分化 TRPV6 MCF10A 3维培养 

分 类 号：Q343[生物学—遗传学]