[1]王娇娇),范智蕊),李砺锋),等.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].郑州大学学报(医学版),2017,(05):536-540.[doi:10.13705/j.issn.1671-6825.2017.05.005]
 WANG Jiaojiao),FAN Zhirui),LI Lifeng),et al.Expression of HPK1 in breast carcinoma tissue and effects of HPK1 overexpression on proliferation and apoptosis of human breast carcinoma cell MCF-7[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2017,(05):536-540.[doi:10.13705/j.issn.1671-6825.2017.05.005]
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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*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2017年05期
页码:
536-540
栏目:
论著
出版日期:
2017-09-15

文章信息/Info

Title:
Expression of HPK1 in breast carcinoma tissue and effects of HPK1 overexpression on proliferation and apoptosis of human breast carcinoma cell MCF-7
作者:
王娇娇1)范智蕊2)李砺锋23)丁显飞24)周学良2)杨子悦4)袁 博24)许振涛24)马丙钧5)赵 杰5)王留兴2)#
1)郑州大学第一附属医院超声科 郑州 450052 2)郑州大学第一附属医院肿瘤科 郑州450052 3)郑州大学第一附属医院生物细胞治疗中心 郑州 450052 4)郑州大学第一附属医院综合ICU 郑州 450052 5)郑州大学第一附属医院药学部 郑州 450052
Author(s):
WANG Jiaojiao1) FAN Zhirui2) LI Lifeng23) DING Xianfei24) ZHOU Xueliang2) YANG Ziyue4) YUAN Bo24) XU Zhentao24) MA Bingjun5) ZHAO Jie5) WANG Liuxing2)
1)Department of Ultrasound, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 2)Department of Oncology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 3)Biotherapy Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 4)Department of General ICU, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 5)Department of Pharmacy, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052
关键词:
HPK1 过表达 乳腺癌 MCF-7细胞 增殖 凋亡
Keywords:
HPK1 overexpression breast carcinoma MCF-7 cell proliferation apoptosis
分类号:
R737.9
DOI:
10.13705/j.issn.1671-6825.2017.05.005
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pCDH-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 kinase 1(HPK1)in carcinoma tissue of human invasive ductal breast carcinoma-not otherwise specified and breast carcinoma cells, and to investigate the effects of HPK1 overexpression on proliferation and apoptosis of human breast carcinoma cells by constructing HPK1 lentiviral vector.Methods: The expression level of HPK1 protein in carcinoma tissue and paired adjacent tissue from 48 cases of breast carcinoma was detected by Western blot. The expression level of HPK1 in MCF10A and MCF-7 cells was detected by Western blot and RT-PCR. The HPK1 sequence was amplified by PCR, lentivirus recombinant vector pCDH-HPK1-puro was constructed, virus was packaged and MCF-7 cells were transfected(overexpression group). The MCF-7 cells infected with empty vector virus was used as control group. The expression level of HPK1 protein and mRNA was detected by Western blot and RT-PCR, respectively. The cell proliferation ability, cell apoptosis and cell cycle were detected by MTT assay and flow cytometry, respectively.Results: The expression level of HPK1 protein was lower in breast carcinoma tissue compared with adjacent tissue(P=0.036). The level of HPK1 protein and mRNA was lower in MCF-7 cells, compared with that of MCF10A cells(P<0.05). Compared with the control group, the expression level of HPK1 in the overexpression group was up-regulated, cell proliferation ability was decreased significantly, while the apoptosis rate and the proportion of G0/G1 were significantly increased(P<0.05).Conclusion: The expression of HPK1 in both breast carcinoma tissue and cell line is low, and overexpression of HPK1 could inhibit proliferation and induce apoptosis and block the cells in G0/G1 phase.

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备注/Memo

备注/Memo:
*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
#通信作者,男,1952年2月生,本科,教授,主任医师,研究方向:乳腺癌和前列腺癌的基础与临床,E-mail:lxwang2006@126.com
更新日期/Last Update: 2017-09-20