siRNA下调Ezrin表达对子宫内膜癌症HEC-1B细胞凋亡和增殖的影响

中国综合临床 Pub Date : 2019-05-01 DOI:10.3760/CMA.J.ISSN.1008-6315.2019.03.014

Hua Chang, Xue Zhang, Baixue Li, K. Guo

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Guo","doi":"10.3760/CMA.J.ISSN.1008-6315.2019.03.014","DOIUrl":null,"url":null,"abstract":"Objective \nTo observe the changes of cell cycle, apoptosis and proliferation of endometrial cancer cells after the expression and down-regulation of Ezrin in endometrial cancer cells and to explore whether Ezrin may be a candidate gene for targeted therapy. \n \n \nMethods \nEndometrial cancer cells were from Shanghai Institute of Cell Research, of Chinese Academy of Medical Sciences in February 2017 and divided into blank control group and siEzrin group according to the intervention methods.Western blot and qRT-PCR was used to detect the expression of Ezrin protein and mRNA in endometrial cell lines.Small interfering RNA(siRNA) was used to transfect HEC-1B cell and down-regulate Ezrin.Cell cycle and apoptosis were detected by flow cytometry.MTT assay was used to detect multiplication. \n \n \nResults \nWestern blot showed that Ezrin protein was expressed in Ishikawa(31.742±5.832)、HEC-1A(16.326±3.135)、HEC-1B(17.636±4.426) and KLE(14.862±5.109) and qRT-PCR showed that mRNA was expressed in Ishikawa (2.513±0.725), HEC-1A (1.655±0.692), HEC-1B (3.237±0.411) and KLE (0.962±0.235) cell lines, and expressed highest in HEC-1B cells (F=6.173, P<0.05; F=7.042, P<0.05). 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引用次数: 0

摘要

目的观察Ezrin基因在子宫内膜癌细胞中表达及下调后对子宫内膜癌细胞周期、凋亡及增殖的影响，探讨Ezrin基因是否可作为靶向治疗的候选基因。方法2017年2月取自中国医学科学院上海细胞研究所的子宫内膜癌细胞，按干预方法分为空白对照组和siEzrin组。采用Western blot和qRT-PCR检测Ezrin蛋白和mRNA在子宫内膜细胞系中的表达。小干扰RNA(siRNA)转染HEC-1B细胞，下调Ezrin。流式细胞术检测细胞周期和凋亡情况。MTT法检测增殖。结果Western blot结果显示，Ezrin蛋白在Ishikawa(31.742±5.832)、HEC-1A(16.326±3.135)、HEC-1B(17.636±4.426)和KLE(14.862±5.109)细胞中均有表达，qRT-PCR结果显示，mRNA在Ishikawa(2.513±0.725)、HEC-1A(1.655±0.692)、HEC-1B(3.237±0.411)和KLE(0.962±0.235)细胞中均有表达，且在HEC-1B细胞中表达量最高(F=6.173, P<0.05;F = 7.042, P < 0.05)。流式细胞术检测结果显示，与空白对照组相比，G1期和G2期细胞较少，S期细胞较多(t=3.118, P<0.05);t = 5.435, P < 0.05;t = 3.332, P < 0.05)。下调Ezrin后，HEC-1B细胞的凋亡率由(9.84±2.37)%升高至(17.64±5.96)%(t=8.963, P<0.01)。MTT检测显示，转染siEzrin 72 h和96 h后HEC-1B细胞的增殖均低于空白对照组(t=3.209, P<0.05;t = 3.726, P < 0.05)。结论下调Ezrin可使更多的子宫内膜癌细胞停留在S期，促进细胞凋亡，抑制细胞增殖，Ezrin可能成为靶向治疗的靶标候选基因。关键词:子宫内膜癌;Ezrin蛋白;细胞周期;细胞凋亡;扩散

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Effect of apoptosis and proliferation of endometrial cancer HEC-1B cell by siRNA down-regulating expression of Ezrin

Objective To observe the changes of cell cycle, apoptosis and proliferation of endometrial cancer cells after the expression and down-regulation of Ezrin in endometrial cancer cells and to explore whether Ezrin may be a candidate gene for targeted therapy. Methods Endometrial cancer cells were from Shanghai Institute of Cell Research, of Chinese Academy of Medical Sciences in February 2017 and divided into blank control group and siEzrin group according to the intervention methods.Western blot and qRT-PCR was used to detect the expression of Ezrin protein and mRNA in endometrial cell lines.Small interfering RNA(siRNA) was used to transfect HEC-1B cell and down-regulate Ezrin.Cell cycle and apoptosis were detected by flow cytometry.MTT assay was used to detect multiplication. Results Western blot showed that Ezrin protein was expressed in Ishikawa(31.742±5.832)、HEC-1A(16.326±3.135)、HEC-1B(17.636±4.426) and KLE(14.862±5.109) and qRT-PCR showed that mRNA was expressed in Ishikawa (2.513±0.725), HEC-1A (1.655±0.692), HEC-1B (3.237±0.411) and KLE (0.962±0.235) cell lines, and expressed highest in HEC-1B cells (F=6.173, P<0.05; F=7.042, P<0.05). Flow cytometry assay showed that compared with blank control group less cells stayed in G1 phase and G2 phase, more stayed in S phase (t=3.118, P<0.05; t=5.435, P<0.05; t=3.332, P<0.05). The apoptotic rate of HEC-1B cells increased from (9.84±2.37)% to (17.64±5.96)%(t=8.963, P<0.01) after Ezrin was down-regulated.MTT assay showed that the proliferation of HEC-1B cells in 72 h and 96 h siEzrin transfection group was lower than that in blank control group (t=3.209, P<0.05; t=3.726, P<0.05). Conclusion Down-regulating of Ezrin may promote more endometrial cancer cells stay in S phase and promote apoptosis, inhibit proliferation, Ezrin may become target candidate gene in target therapy. Key words: Endometrial cancer; Ezrin protein; Cell cycle; Apoptosis; Proliferation

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来源期刊

中国综合临床

CiteScore

0.10

自引率

0.00%

发文量

16855

期刊介绍： Clinical Medicine of China is an academic journal organized by the Chinese Medical Association (CMA), which mainly publishes original research papers, reviews and commentaries in the field. Clinical Medicine of China is a source journal of Peking University (2000 and 2004 editions), a core journal of Chinese science and technology, an academic journal of RCCSE China Core (Extended Edition), and has been published in Chemical Abstracts of the United States (CA), Abstracts Journal of Russia (AJ), Chinese Core Journals (Selection) Database, Chinese Science and Technology Materials Directory, Wanfang Database, China Academic Journal Database, JST Japan Science and Technology Agency Database (Japanese) (2018) and other databases.