| Prohibitin 表达及对乳腺癌细胞系MCF-7 抑制生长的机制研究 |
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| 引用本文： | 周晓艳,李 越,何 谦,袁晓红,张海宝.Prohibitin 表达及对乳腺癌细胞系MCF-7 抑制生长的机制研究[J].现代检验医学杂志,2020,0(3):6-10. |
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| 作者姓名： | 周晓艳 李 越 何 谦 袁晓红 张海宝 |
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| 作者单位： | （1. 西安交通大学第二附属医院检验科，西安 710004；
2. 西安交通大学环境与疾病相关基因教育部重点实验室，西安 710061） |
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| 摘 要： | 目的 该研究从细胞水平研究了抗增殖蛋白（prohibitin）高表达在乳腺癌细胞中的生物学功能，并研究其抑制
MCF-7 生长的分子机制。方法 使用基因重组法构建pEGFP-PHB 重组质粒，脂质体瞬时转染乳腺癌细胞系MCF-7，
real time-PCR 和Western Blotting 验证其高表达后，检测细胞生物学功能。包括:MTT（噻唑蓝）法检测细胞生长、流
式细胞仪检测细胞周期与凋亡、肿瘤相关基因P53, Bcl-2, ERBB2 及E2F-1 的检测和Transwell 小室侵袭实验。两样本差
异比较采用t 检验，多样本差异采用单因素方差分析。结果 MTT 显示PHB 高表达组在转染24, 48 和72h 对细胞生长
的抑制率分别为20.98%, 14.93% 和62.94%，差异有统计学意义；流式细胞仪检测细胞分裂周期，phb 高表达G1 期无差异，
S 期降低，G2 期增加；凋亡检测中，高表达组细胞总凋亡率为33.67%±8.68%，阴性对照组的凋亡率12.13%±3.76%，
空白对照组凋亡率为6.99%±2.33%，高表达组细胞凋亡率高于对照组，差异有统计学意义（F=18.992, P=0.003）；
PHB 高表达时，肿瘤相关蛋白P53 和ERBB2 的表达量高于对照组（t=4.590, P=0.044; t=9.489, P=0.011），Bcl-2 表达降
低，差异有统计学意义(t=7.143, P=0.019)，E2F-1 蛋白各组间差异无统计学意义(t=2.175, P=0.162)；PHB 高表达组细胞
侵袭能力低于对照组，差异有统计学意义（t=3.221, P=0.01;t=4.057, P=0.003）。结论 PHB 高表达抑制S 期DNA 的合
成，阻滞细胞于G2/M 期，增加细胞凋亡率，抑制肿瘤细胞MCF-7 的生长。其与肿瘤相关蛋白P53 和ERBB2 的增高，
Bcl-2 表达的降低紧密相关。
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| 关 键 词： | 抗增殖蛋白 乳腺癌 增殖 凋亡 |
Expression of Prohibitin and Its Restrain Mechanism to Growth
in the Breast Cancer Cell MCF-7 |
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| ZHOU Xiao-yan,LI Yue,HE Qian,YUAN Xiao-hong,ZHANG Hai-bao.Expression of Prohibitin and Its Restrain Mechanism to Growth
in the Breast Cancer Cell MCF-7[J].Journal of Modern Laboratory Medicine,2020,0(3):6-10. |
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| Authors: | ZHOU Xiao-yan LI Yue HE Qian YUAN Xiao-hong ZHANG Hai-bao |
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| Affiliation: | （1. Department of Clinical Laboratory, the Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004,
China; 2. Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education,
Xi’an Jiaotong University, Xi’an 710061, China ） |
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| Abstract: | Objective To explore the biological function of PHB in breast cancer cells from the level of molecular cell and the
mechanism of restrain to MCF-7. Methods After construction of pEGFP-PHB recombinant plasmid, it was transfected into
breast cancer cell line MCF-7 by liposome and verified prohibitin high expression with RT-PCR and western blotting latter. The
biological function of PHB in MCF-7 cells was detected by a series of experiments. The biological functions of the cells were
detected, including MTT (thiazole blue) method for cell growth, flow cytometry for cell cycle and apoptosis, detectiion of tumor
related genes P53,bcl-2 ERBB2 and E2F-1, and Transwell’s compartment invasion experiment t test was used in the mean of the
two samples, and variance analysis was used for the diversity. Results MTT showed that the inhibition rate of cell growth was
20.98%, 14.93% and 62.94% in the high expression group of PHB with 24, 48 and 72h, respectively. Flow cytometry was used to
detect the cell intersecting period, and there was no difference in the G1 phase of PHB high expression group compared with
controls, and the period of S phase decreased and the G2 phase increased. Apoptosis detection, OE group of apoptosis rate was
33.67% ± 8.68 %, apoptosis rate of the negative control group 12.13% ± 3.76 %, the apoptosis rate of blank control group was
6.99%± 2.33%, group high expression of apoptosis rate was higher than the control group, and the difference was statistically significant (F=18.992, P=0.003). When PHB was high expressed, P53 and ERBB2 were higher than those in the control group
(t=4.590, P=0.044; t=9.489, P=0.011), the expression of Bcl-2 in over-expression group decreased, and the difference was
statistically significant (t=7.143, P=0.019), and there was no difference between the groups of E2F-1 protein (t=2.175, P=0.162).
The cell invasion ability of PHB high expression group was higher than that of control group, and the difference was statistically
significant (t=3.221, P=0.01). Conclusion The high expression of PHB inhibited the synthesis of S-phase DNA, blocked the
cells in G2/M phase, increased the rate of apoptosis, and inhibited the growth of tumor cells MCF-7. It was closely related to the
increase of tumor-related proteins P53 and ERBB2, and the decrease of bcl-2 expression. |
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