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

发布时间：2018-06-29 03:13

本文选题：HPK + 过表达　；参考：《郑州大学学报(医学版)》2017年05期

【摘要】：目的:检测造血祖细胞激酶1(HPK1)在人非特异型浸润性乳腺癌组织及乳腺癌细胞中的表达,并构建HPK1慢病毒载体以探究HPK1过表达对人乳腺癌细胞增殖、凋亡的影响。方法:采用Western blot检测48例乳腺癌组织和配对癌旁组织中HPK1蛋白的表达,采用Western blot、RT-PCR法检测正常乳腺上皮细胞(MCF10A)和乳腺癌MCF-7细胞中HPK1的表达水平。PCR扩增HPK1序列,构建慢病毒p CDH-HPK1-puro重组载体,包装病毒、转染MCF-7细胞(过表达组),以感染空载体病毒的MCF-7细胞作为对照组,分别采用Western blot、RT-PCR检测2组细胞HPK1的表达水平,MTT法检测细胞增殖能力的变化,流式细胞术检测细胞凋亡及细胞周期。结果:与癌旁组织相比,HPK1蛋白在乳腺癌组织中低表达(P=0.036)。与MCF10A细胞相比,HPK1蛋白及mRNA在MCF-7细胞中低表达;与对照组相比,过表达组中HPK1蛋白及mRNA的表达水平上调,细胞增殖能力下降,细胞凋亡率升高,G0/G1期细胞比例升高(P均0.05)。结论:HPK1在乳腺癌组织及细胞中低表达,HPK1过表达可抑制MCF-7细胞增殖和诱导凋亡,并使细胞周期阻断在G0/G1期。
[Abstract]:Aim: to detect the expression of hematopoietic progenitor cell kinase 1 (HPK1) in human non-heterogenous invasive breast cancer and breast cancer cells, and construct HPK1 lentivirus vector to investigate the effect of HPK1 overexpression on proliferation and apoptosis of human breast cancer cells. Methods: the expression of HPK1 protein was detected by Western blot in 48 breast cancer tissues and matched adjacent tissues. The expression level of HPK1 in normal breast epithelial cells (MCF10A) and breast cancer MCF-7 cells was detected by RT-PCR. The recombinant vector of lentivirus pCDH-HPK1-puro was constructed, and the vector was packaged and transfected into MCF-7 cells (overexpression group). MCF-7 cells infected with empty vector virus were used as control group. The expression level of HPK1 in the two groups was detected by Western blottin RT-PCR and the cell proliferation was detected by MTT assay. Apoptosis and cell cycle were detected by flow cytometry. Results: the expression of HPK1 protein in breast cancer tissues was lower than that in paracancerous tissues (P0. 036). Compared with MCF10A cells, the expression of HPK1 protein and mRNA in MCF-7 cells was lower than that in MCF10A cells, and the expression level of HPK1 protein and mRNA in overexpression group was up-regulated, the cell proliferation ability was decreased, and the percentage of HPK1 protein and mRNA was increased in G0 / G1 phase (P 0.05). Conclusion the overexpression of HPK1 in breast cancer tissues and cells can inhibit the proliferation and induce apoptosis of MCF-7 cells and block the cell cycle in G _ 0 / G _ 1 phase.
【作者单位】：郑州大学第一附属医院超声科;郑州大学第一附属医院肿瘤科;郑州大学第一附属医院生物细胞治疗中心;郑州大学第一附属医院综合ICU;郑州大学第一附属医院药学部;
【基金】：科技部科技惠民计划专项基金资助项目2013GS410101 河南省重大科技专项基金资助项目151100310800
【分类号】：R737.9

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