贵州医科大学学报

2019, v.44;No.226(07) 792-797

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PKA抑制剂在雌激素调控乳腺癌细胞纤连蛋白表达中的作用及机制
The Role and Mechanism of PKA Inhibitors in the Expression of Fibronectin of Estrogen-induced Breast Cancer Cells

王宏健;刘晓红;董宇华;郭阳;王旭东;
WANG Hongjian;LIU Xiaohong;DONG Yuhua;GUO Yang;WANG Xudong;Department of Physiology,Guizhou Medical University;

摘要(Abstract):

目的:观察蛋白激酶A(PKA)的抑制剂H89在雌激素(E2)依赖人乳腺癌细胞(MCF-7)纤连蛋白(fibronectin,FN)表达及其与CANP2的关系。方法:分别用E2和PKA抑制剂H89刺激对数生长期MCF-7细胞和CANP2-shRNA转染MCF-7模型细胞,采用伤口愈合实验检测细胞迁移率、Western-blot法测FN蛋白的表达。结果:与对照组比较,H89预处理后,E2诱导的MCF-7细胞迁移率可明显增加,E2与H89联用可使FN蛋白表达明显上调,差异有统计学意义(P<0.05);H89和E2+H89组的可促使CANP2-shRNA转染MCF-7细胞的迁移率明显降低,并抑制CANP2-shRNA转染MCF-7细胞中FN蛋白表达,差异有统计学意义(P<0.01)。结论:PKA抑制剂能明显增强E2诱导的乳腺癌细胞迁移和FN蛋白表达,而该效应可被CANP2基因沉默所逆转。
Objective:To observe the impact of H89 on the cell migration and the protein expression of fibronectin(FN) in MCF-7 breast cancer cells and its relationship with calpain2.Methods:Estrogen(E2) and protein kinase A(PKA) inhibitor H89 were used to stimulate MCF-7 cells and CANP2-shRNA transfected model cells in logarithmic growth phase respectively.Cell migration and FN protein expression were measured by wound healing test and Western-blot method.Results:Compared with the control group,the cell migration rate induced by E2 increased significantly after H89 pretreatment(P<0.05),and the expression of FN protein increased significantly after E2 combined with H89(P<0.01).After gene silencing of calcium-activated neutral protease 2(CANP2),the cell migration of H89 group and E2 + H89 group decreased significantly(P<0.01),and the up-regulation of FN protein in H89 group and E2 + H89 group was significantly inhibited(P<0.01 compared with NC group).Conclusion:PKA inhibitors can significantly enhance E2-induced cell migration and FN protein expression in breast cancer cells,which can be reversed by CANP2 gene silencing.

关键词(KeyWords): 乳腺肿瘤;雌激素类;蛋白酶抑制剂;纤连蛋白类;细胞迁移;钙蛋白酶2
breast neoplasms;estrogen;protease inhibitors;fibronectin;cell migration;calpain 2

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅-贵州医科大学科技合作计划项目[黔科合LH(2015)7317];; 国家自然科学基金项目(31360252,31660345)

作者(Author): 王宏健;刘晓红;董宇华;郭阳;王旭东;
WANG Hongjian;LIU Xiaohong;DONG Yuhua;GUO Yang;WANG Xudong;Department of Physiology,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2019.07.010

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