Pten对奶牛乳腺上皮细胞活力及乳蛋白合成的影响  

Impact of Pten on Cell Viability and Milk Protein Synthesis in Dairy Cow Mammary Epithelial Cells

作　　者：王卓然[1] 侯晓明[1] 曲波[1] 王杰[1] 高学军[1] 李庆章[1] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室,哈尔滨150030

出　　处：《中国生物化学与分子生物学报》2014年第4期395-401,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展规划(973计划;No.2011CB100804)~~

摘　　要：Pten作为抑癌基因,参与调控细胞生长、粘附、凋亡以及其它细胞活动.目前,国内外关于Pten在奶牛乳腺发育过程中表达及调节的研究鲜有报道.为了揭示Pten的表达与奶牛乳腺发育与泌乳之间的关系,本研究应用qRT-PCR技术检测Pten在不同泌乳时期和不同乳品质的奶牛乳腺组织中的表达差异,进而应用脂质体转染方法,通过siRNA介导的RNA干扰技术改变Pten基因在奶牛乳腺上皮细胞中的表达量,CASY法检测细胞活力,用ELISA试剂盒检测细胞分泌β-酪蛋白的含量,采用qRT-PCR、Western印迹等技术检测Pten对奶牛乳腺上皮细胞中乳蛋白相关信号通路基因表达的影响.结果显示,泌乳期高乳品质奶牛乳腺组织中Pten表达水平显著低于泌乳期低乳品质及干乳期奶牛;Pten基因沉寂后,细胞活力提高,β-酪蛋白质量浓度增加,CSN2、AKT、MTOR、STAT5表达量增加.研究表明,Pten可通过抑制细胞活力和乳蛋白分泌而影响泌乳.Pten, as a tumor suppressor gene, participates in the regulation of cell growth, adhesion, apoptosis and other cell progresses. However, the knowledge about expression and regulation mechanism of Pten in the development of dairy cow mammary gland is limited. To reveal the relationship among Pten and dairy cow mammary gland development and lactation, qRT-PCR was used to analyze the expression level of Pten of dairy cow mammary gland in different lactation period and milk quality. Liposome transfection technique was used to change the expression of Pten in dairy cow mammary epithelial cells by RNA interference. The viability of dairy cow mammary epithelial ceils was analyzed by CASY-YY method. The content of 13-casein was detected by ELISA. The effect of Pten on milk protein related signaling pathway genes in dairy cow mammary epithelial cells was analyzed by qRT-PCR and Western blotting, respectively. The results showed that Pten mRNA expression was decreased significantly in lactation mammary gland with high milk quality, compared with lactation mammary gland with low milk quality and pregnancy period tissue. After Pten silence cell viability was enhanced, β-casein content was increased, and CSN2, AKT, MTOR, STAT5 expression was increased also. The results suggest that Pten may inhibit mammary epithelial cells lactation by suooressinz cell viability and milk nrotain svntha^i~

关 键 词：PTEN RNA干扰 奶牛乳腺 细胞活力 乳蛋白 

分 类 号：Q527[生物学—生物化学]