[1]周晓艳,李　越,何　谦,等.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,2020,35(03):6-10.[doi:10.3969/j.issn.1671-7414.2020.03.002]
　ZHOU Xiao-yan,LI Yue,HE Qian,et al.Expression of Prohibitin and Its Restrain Mechanism to Growth in the Breast Cancer Cell MCF-7[J].Journal of Modern Laboratory Medicine,2020,35(03):6-10.[doi:10.3969/j.issn.1671-7414.2020.03.002]
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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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《现代检验医学杂志》[ISSN:/CN:]

卷:
第35卷
期数:
2020年03期
页码:
6-10
栏目:
论著
出版日期:
2020-07-20

文章信息/Info

Title:
Expression of Prohibitin and Its Restrain Mechanism to Growth in the Breast Cancer Cell MCF-7
文章编号:
1671-7414（2020）03-006-06
作者:
周晓艳1; 李　越1; 何　谦1; 袁晓红1; 张海宝2
（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）
Author(s):
ZHOU Xiao-yan1;  LI Yue1;  HE Qian1;  YUAN Xiao-hong1;  ZHANG Hai-bao2
（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 ）
关键词:
抗增殖蛋白; 乳腺癌; 增殖; 凋亡
分类号:
R737.9；R730.43
DOI:
10.3969/j.issn.1671-7414.2020.03.002
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。
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.

参考文献/References:

[1]　 MISHRA S, NYOMBA B G. Prohibitin - At the crossroads of obesity-linked diabetes and cancer[J]. Exp Biol Med，2017, 242(11): 1170-1177.
[2]　 PENG Yating, CHEN Ping , OUYANG Ruoyun, et al. Multifaceted role of prohibitin in cell survival and apoptosis[J]. Apoptosis, 2015, 20(9):1135-1149.
[3]　 TSUTSUMI T, MATSUDA M, AIZAKI H, et al. Proteomics analysis of mitochondrial proteins reveals overexpression of a mitochondrial protein chaperon, prohibitin, in cells expressing hepatitis C virus core protein[J]. Hepatology, 2010,50(2): 378-386.
[4]　 THEISS, A L, Sitaraman S V. The role and therapeutic potential of prohibitin in disease[J]. Biochimica et Biophysica Acta (BBA)/Molecular Cell Research, 2011, 1813(6):1137-1143.
[5]　 THUAUD, F, RIBEIRO N, NEBIGIL C G, et al. Prohibitin ligands in cell death and survival: mode of action and therapeutic potential[J]. Chem Biol, 2013， 20(3): 316-331.
[6]　 HE Qian, ZHANG Shuqun, CHU Yonglie, et al. Separation and identification of differentially expressed nuclear matrix proteins in breast carcinoma forming[J]. Acta Oncologica, 2010, 49(1): 76-84.
[7]　　 赵丽华, 何谦, 张淑群, 等. prohibitin 在乳腺癌 MCF-7 细胞表达抑制的鉴定[J]. 现代检验医学杂志, 2011, 26(6):17-20. 　　 ZHAO Lihua, HE Qian, ZHANG Shuqun, et al. Effect of shRNA expression vectors for Prohibitin transfected MCF-7 cell in vitro[J]. Journal of Modern Laboratory Medicine, 2011, 26(6):17-20.
[8]　　 KAHL A, ANDERSON C J , Qian Liping, et al. Neuronal expression of the mitochondrial protein prohibitin confers profound neuroprotection in a mouse model of focal cerebral ischemia[J]. Journal of Cerebral Blood Flow & Metabolism,2017:38(6): 271678X17720371.
[9]　　 CIRILO P D R, DE SOUSA ANDRAPE L N, CORREA B R S, et al. MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1[J]. BMC Cancer, 2017, 17(1): 750.
[10] KIM N H, YOSHIMARU T, Chen Y A, et al. BIG3 inhibits the estrogen-dependent nuclear translocation of PHB2 via multiple karyopherin-alpha proteins in breast cancer Cells [J]. PLOS One,2015,10(6): e127707.
[11] EL-ETREBY N M, GHAZY A A, RASHAD R, et al. Prohibitin: targeting peptide coupled to ovarian cancer, luteinization and TGF-β pathways[J]. Journal of Ovarian Research, 2017, 10(1)：28.
[12] KOUSHYAR S, ECONOMIDES G, ZAAT S, et al. The prohibitin-repressive interaction with E2F1 is rapidly inhibited by androgen signalling in prostate cancer cells [J]. Oncogenesis, 2017, 6(5): e333.
[13] CHOWDHURY I, THOMPSON W E, WELCH C, et al. Prohibitin (PHB) inhibits apoptosis in rat granulosa cells (GCs) through the extracellular signal-regulated kinase 1/2 (ERK1/2) and the Bcl family of proteins [J]. Apoptosis，2013, 18(12): 1513-1525.
[14] CHOWDHURY I, THOMPSON W E, THOMAS K, et al. Prohibitins role in cellular survival through Ras-Raf- MEK-ERK pathway[J]. J Cell Physiol,2014,229(8): 998-1004.

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备注/Memo

备注/Memo:
收稿日期：2019-11-26 修回日期：2020-01-02基金项目：陕西省国际科技合作重点项目（2012KW-35）。 作者简介：周晓艳（1990-），女，硕士研究生，检验技师，主要从事肿瘤功能学研究，E-mail：xiaoyanzhou10@163.com。 通讯作者：何谦（1971-）, 女，主任技师，主要从事生化和分子生物学研究。
更新日期/Last Update: 2020-07-20