HPK1过表达对乳腺癌MCF-7和MDA-MB-231细胞增殖和凋亡的影响及其机制

HPK1过表达对乳腺癌MCF-7和MDA-MB-231细胞增殖
和凋亡的影响及其机制
王娇娇;范智蕊;李砺锋;丁显飞;周学良;赵杰;王留兴
【期刊名称】《吉林大学学报（医学版）》
【年(卷),期】2017(043)005
【摘要】Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1
proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1
overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜
0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.%
目的:构建造血祖细胞激酶1 (HPK1)慢病毒载体,探讨HPK1过表达对乳腺癌MCF-7和MDA-MB-231细胞增殖和凋亡的作用,阐明其可能的机制.方法:通过过表达HPK1的慢病毒感染细胞,获得稳定表达HPK1的细胞系(MCF-7-HPK1和MDA-MB-231-HPK1),将2个细胞系分别分为3组,即空白组、对照组(空载体组)和过表
达组;分别采用RT-PCR法和Western blotting法检测乳腺癌细胞中HPK1 mRNA和蛋白表达水平,MTT法检测细胞增殖率,流式细胞术检测细胞周期和凋亡率,Transwell法分析细胞的迁移能力,Western blotting法检测caspase 3、PTEN、MMP-9、MMP-2、Ki-67和HPK1蛋白表达水平.结果:与空白组和对照组比较,乳腺癌MCF-7和MDA-MB-231细胞过表达组中HPK1 mRNA和蛋白表达水平升
高(P＜0.05),细胞增殖率明显降低(P＜0.05),细胞凋亡率明显升高(P＜0.05),穿越Matrigel的细胞数明显减少(P＜0.05),MCF-7细胞周期阻断在G1期;过表达组细
胞中caspase 3、PTEN蛋白表达水平上调(P＜0.05),MMP-9、MMP-2、Ki-67
蛋白表达水平降低(P＜0.05).结论:HPK1过表达可抑制乳腺癌细胞MCF-7和
MDA-MB-231的增殖及迁移并诱导凋亡,其可能与caspase 3、PTEN蛋白的表达上调及MMP-9、MMP-2和Ki-67蛋白的表达降低有关.
【总页数】9页(P910-917,后插4)
【作者】王娇娇;范智蕊;李砺锋;丁显飞;周学良;赵杰;王留兴
【作者单位】郑州大学第一附属医院超声科,河南郑州450052;郑州大学第一附属医院肿瘤科,河南郑州450052;郑州大学第一附属医院生物细胞治疗中心,河南郑州450052;郑州大学第一附属医院综合ICU,河南郑州450052;郑州大学第一附属医院肿瘤科,河南郑州450052;郑州大学第一附属医院药学部,河南郑州450052;郑州大学第一附属医院肿瘤科,河南郑州450052
【正文语种】中文
【中图分类】R737.9
【相关文献】
1.Bcl-3在人乳腺癌组织中的表达及对乳腺癌MDA-MB-231细胞增殖与凋亡的影响 [J], 袁建良;胡永伟;丁厚中
2.HPK1在乳腺癌组织中的表达及HPK1过表达对人乳腺癌MCF-7细胞增殖、凋亡的影响 [J], 王娇娇;赵杰;王留兴;范智蕊;李砺锋;丁显飞;周学良;杨子悦;袁博;许振涛;马丙钧
3.TM4SF1在人乳腺癌细胞 MCF-7和 MDA-MB-231中的表达及其对细胞增殖、迁移能力的影响 [J], 张卫芳;孙燕;张伟杰;朱换;王留兴
4.姜黄素对乳腺癌MCF-7、MDA-MB-231细胞增殖、侵袭的影响及其机制探讨[J], 孙永;方泰惠;周静;周鸣鸣;徐斌
5.疏肝益肾方联合内分泌药物对人乳腺癌MCF-7、MDA-MB-231细胞增殖、凋亡的影响 [J], 郝满霞;王希瑞;李伟汉
因版权原因，仅展示原文概要，查看原文内容请购买。