转化生长因子-β1及p21对乳腺癌干细胞的作用
The research of the transforming growth factor β1 and p21 effect on breast cancer stem cells
目的 观察转化生长因子-β1 (TGF-β1)诱导乳腺癌耐阿霉素细胞株MCF7/ADM中乳腺癌干细胞富集的作用,比较乳腺癌干细胞和MCF7/ADM中 p21 基因和蛋白水平的表达差异.方法 流式细胞仪检测阿霉素耐药乳腺癌细胞(ADM细胞)、乳腺细胞球培养法富集的乳腺癌细胞(SC细胞)及加入TGF-β1细胞因子诱导的乳腺癌细胞(SC-T细胞)中乙醛脱氢酶1(ALDH1)阳性细胞比例;比较上述3组细胞微球形成能力差异;逆转录-聚合酶链反应(RT-PCR)及Western blot检测3组细胞中p21基因及蛋白水平的表达差异.结果 ADM组、SC组及SC-T组中ALDH1阳性细胞比例分别为8.49%、14.94%及23.44%;细胞微球形成率分别为5.29%、11.67%、23.13%;3组细胞中p21 mRNA及蛋白水平的表达均逐渐升高.结论TGF-β1能诱导乳腺癌干细胞的富集并使其具有更强的自我更新能力.P21在乳腺癌干细胞中高表达.
更多Objective To detect the enrichment of breast cancer stem cells induced by transforming growth factor β1 (TGF-β1),and the difference of p21 expression between breast cancer stem cells and MCF7/ADM cells.Methods Flow cytometry detect the proportion of aldehyde dehydrogenase Ⅰ (ALDH1) positive cells in MCF7/ADM,breast cancer stem cells enriched by mammosphere and breast cancer stem cells induced by TGF-β1.To detect the mammoshpore formation ability in above three teams by mammoshpere formation test.To detect the p21 expression in three teams above by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.Results The proportion of ALDH1 positive cells in the three teams were 8.49%,14.94%,23.44% respectively.The mammosphere formation rate were 5.29%,11.67%,23.13% respectively.The expression of p21 increased in the three teams.Conclusion TGF-β1 can induce the enrichment of breast cancer stem cells,and enhance it' s self-renewal ability.The expression of p21 is higher in breast cancer stem cells than common breast cancer cells.
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