代谢型谷氨酸受体4的真核表达及其在乳腺癌细胞增殖中的作用  

Eukaryotic Expression of mGluR4 and its Role in the Proliferation of Breast Cancer Cell

作　　者：肖斌[1] 陈丽丹[1] 孙朝晖[1] 廖扬[1] 杭建峰[1] 陈建芸[1] 张蓉[1] 李林海[1] 

机构地区：[1]广州军区广州总医院检验科,广东广州510010

出　　处：《生物技术通讯》2017年第2期109-113,共5页Letters in Biotechnology

基　　金：2015年广州市科创委项目(201604040003)

摘　　要：目的:建立代谢型谷氨酸受体4(mGluR4)在乳腺癌细胞MCF-7中高表达的模型,探索mGluR4的亚细胞定位及其在乳腺癌增殖中的作用。方法:提取MCF-7细胞总RNA并逆转录为cDNA;PCR扩增mGluR4基因;利用无缝克隆技术快速构建以pENTER为载体的mGluR4重组表达质粒;通过Western印迹和免疫荧光实验检测mGluR4在MCF-7细胞中的表达及定位;利用CCK8实验观察mGluR4对MCF-7细胞增殖的影响。结果:构建了能够在MCF-7细胞中高表达的mGluR4真核表达质粒,免疫荧光结果显示mGluR4定位于细胞膜和细胞质,CCK8实验表明高表达mGluR4显著促进乳腺癌细胞的增殖能力。结论:为探索mGluR4的相关功能制备了必要工具,为研究mGluR4在乳腺癌增殖中的分子机制奠定了基础。Objective：To express metabotropic glutamate receptor 4（mGluR4）in breast cancer cell line MCF-7,further explore the subcellular localization and the role of mGluR4 in cell proliferation. Methods：The total RNA was extracted from MCF-7 cells and was subsequently reversed into cDNA,and from which,mGluR4 gene was amplified by PCR. Recombinant mGluR4 expression plasmid was constructed using one step cloning kit. Western blotting and immunofluorescence assays were applied to detect the expression and localization of mGluR4 respectively. The effect of mGluR4 on the proliferation of MCF-7 was also measured. Results：The mGluR4 was localized in the cytoplasm and membrane of MCF-7 cells. The expression of mGluR4 could significantly enhance the proliferation of breast cancer cells. Conclusion：This research will provide a useful tool for studying the function of mGluR4 and lay the foundation of the molecular mechanism of mGluR4 in the proliferation of breast cancer.

关 键 词：代谢型谷氨酸受体4 乳腺癌 真核表达 增殖 

分 类 号：Q78[生物学—分子生物学]