Thrsp蛋白在乳腺癌MCF-7、MDA-MB-231细胞株中表达及鉴定

Expression and identification of Thrsp protein in breast cancer MCF-7 and MDA-MB-231 cell lines

目的鉴定Thrsp甲状腺激素应答蛋白(Thrsp,S14)在乳腺癌MCF-7和MDA-MB-231细胞株中的表达,并建立2株细胞的标准生长曲线,为研究甲状腺激素应答蛋白在乳腺癌细胞转移增殖中的作用及机制奠定基础。方法取对数生长期的乳腺癌MCF-7、MDA-MB-231细胞,用CCK8法检测,用Excel自带统计软件求平均值并绘制生长曲线。细胞爬片,Thrsp蛋白免疫组化SP法染色,DBA显色,显微镜下观察,采用Western Blot法检测2株细胞中Thrsp蛋白表达及相对量。结果连续培养72h MCF-7、MDA-MB-231细胞株达到对数生长期。Thrsp蛋白在乳腺癌细胞株MDA-MB-231中呈弱阳性表达,在MCF-7中呈阳性表达。Thrsp蛋白电泳大小位于17ku。MBA-MD-231细胞株中Thrsp蛋白表达水平低于MCF-7细胞株,与免疫组化结果一致。结论 Luminal A乳腺癌细胞株MCF-7细胞株中Thrsp蛋白表达正常;Basal细胞株MDA-MB-231中Thrsp蛋白表达下调。

Objective To identify the expression of thyroid hormone responsive prote（Thrsp）in MCF-7and MDA-MB-231 cell Lines of breast cancer.The standard growth curve of the 2cell line was established.In order to lay the foundation for the founction and mechanism of transfer and proliferation of the breast cancer cell lines by Thrsp protein.Methods To select breast cancer MCF-7,MDA-MB-231 cells in the logarithmic growth phase,detected by CCK8 method,using Excel to calculate the mean value and built the growth curve,the cell climbing piece,Thrsp protein was stained by immunohistochemistry SP method,DBA coloration,observed by microscope,the Western Blot method was used to detect the expression and the relative amount of Thrsp protein in 2cells.Results Continuous culturing for 72 hours,MCF-7and MDA-MB-231 cell line were reaching the logarithmic growth phase.Thrsp protein were expressed weakly positive in MBA-MD-231,and positive in MCF-7.The electrophoresis size of Thrsp protein were 17 ku.The expression of Thrsp protein in MBA-MD-231 were a bit lower than that in MCF-7 which were tested by Western-blot.And the results were consistent with immunohistochemical results.ConclusionThrsp protein was expressed normally in Luminal A breast cancer MCF-7cell line.Thrsp protein was expressed a bit lower in Basal-like cell line MDA-MB-231.