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

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。

Influence of HPK1 overexpression in proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells and its mechanism

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.