YY1对卵巢癌细胞增殖和侵袭作用影响及其分子机制
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摘要
目的探讨Yin Yang 1(YY1)对卵巢癌细胞增殖和侵袭的作用影响及其分子机制。方法选取本院2017年10月至2018年5月收治的85例卵巢癌患者,采用实时定量逆转录聚合酶链式反应(qRT-PCR)和蛋白印迹(Western Blot)分别检测卵巢癌组织(OCT)及其癌旁组织(PT)和正常组织(NT)中YY1的表达;Western Blot检测5种常见的卵巢癌细胞系中YY1的表达;采用siRNA干扰技术下调YY1在卵巢癌细胞系HO8910PM中的表达,qRT-PCR检测干扰前后细胞系YY1的表达;CCK-8和Trans-well检测RNA干扰前后HO8910PM细胞增殖、迁移和侵袭能力的变化;Western Blot分析RNA干扰前后YY1与上皮间充质转化(EMT)标志蛋白之间的变化。结果qRT-PCR和Western Blot检测结果显示,卵巢癌组织中YY1的表达显著高于癌旁组织和正常组织(P<0.05);Western Blot结果显示YY1在HO8910PM细胞系中表达最高;利用siRNA干扰卵巢癌细胞系YY1的表达后,其在HO8910PM表达显著降低(P<0.05);YY1下调后,卵巢癌细胞HO8910PM增殖和侵袭能力及上皮间质转化相关蛋白N-cadherin和Vimentin表达均显著低于未下调YY1的阴性对照组(P<0.05),而E-cadherin的表达显著升高(P<0.05)。结论YY1通过调节EMT机制影响卵巢癌增殖和侵袭能力。
Objective:To investigate the effect of Yin Yang 1(YY1)on proliferation,migration and invasion of ovarian cancer cells and its molecular biology mechanism.Methods:Eighty-five patients with ovarian cancer admitted to our hospital from October 2017 to May2018 were enrolled in this study.The expressions of YY1 in ovarian cancer tissues(OCT)and paracancer tissues(PT)and normal tissues(NT)were detected by using real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot.The expression of YY1 in five ovarian cancer cell lines was detected by using the Western blot.Interfering technique was used to down-regulate the expression of YY1 in ovarian cancer cell line HO8910 PM,and qRT-PCR was used to detect the YY1 expression of cell line before and after interference.The changes in proliferation,migration and invasion of HO8910 PM cells before and after RNA interference were examined by using CCK-8 and Transwell.Western blot was used to analyze the changes of YY1 and epithelial mesenchymal transition(EMT)marker proteins before and after RNA interference.Results:The results of qRT-PCR and Western blot showed that the expression of YY1 in ovarian cancer tissues was significantly higher than in paracancer tissues and normal tissues(P<0.05).Western blot showed that YY1 expression was the highest in HO8910 PM cell line.When siRNA was used to interfere with the YY1 expression of ovarian cancer cell line,the proliferation and invasion ability of ovarian cancer cell line HO8910 PM and the expressions of N-cadherin and vimentin in epithelialmesenchymal transition after YY1 down-regulation were significantly lower than those in the negative control group(P<0.05),while the expression of E-cadherin was significantly increased(P<0.05).Conclusions:YY1 affects the proliferation,migration and invasion of ovarian cancer by regulating the epithelial mesenchymal transition.
Objective:To investigate the effect of Yin Yang 1(YY1)on proliferation,migration and invasion of ovarian cancer cells and its molecular biology mechanism.Methods:Eighty-five patients with ovarian cancer admitted to our hospital from October 2017 to May2018 were enrolled in this study.The expressions of YY1 in ovarian cancer tissues(OCT)and paracancer tissues(PT)and normal tissues(NT)were detected by using real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot.The expression of YY1 in five ovarian cancer cell lines was detected by using the Western blot.Interfering technique was used to down-regulate the expression of YY1 in ovarian cancer cell line HO8910 PM,and qRT-PCR was used to detect the YY1 expression of cell line before and after interference.The changes in proliferation,migration and invasion of HO8910 PM cells before and after RNA interference were examined by using CCK-8 and Transwell.Western blot was used to analyze the changes of YY1 and epithelial mesenchymal transition(EMT)marker proteins before and after RNA interference.Results:The results of qRT-PCR and Western blot showed that the expression of YY1 in ovarian cancer tissues was significantly higher than in paracancer tissues and normal tissues(P<0.05).Western blot showed that YY1 expression was the highest in HO8910 PM cell line.When siRNA was used to interfere with the YY1 expression of ovarian cancer cell line,the proliferation and invasion ability of ovarian cancer cell line HO8910 PM and the expressions of N-cadherin and vimentin in epithelialmesenchymal transition after YY1 down-regulation were significantly lower than those in the negative control group(P<0.05),while the expression of E-cadherin was significantly increased(P<0.05).Conclusions:YY1 affects the proliferation,migration and invasion of ovarian cancer by regulating the epithelial mesenchymal transition.
引文
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[12]Huang T,Wang G,Yang L,et al.Transcription factor YY1modulates lung cancer progression by activating lncRNA-PVT1[J].DNA Cell Biol,2017,36:947-958.
[13]Balkhi MY,Wittmann G,Xiong F,et al.YY1 Upregulates checkpoint receptors and downregulates type I cytokines in exhausted,chronically stimulated human T cells[J].iScience,2018,27:105-122.
[14]Lu Z,Hong CC,Kong G,et al.Polycomb group protein YY1is an essential regulator of hematopoietic stem cell quiescence[J].Cell Rep,2018,22:1545-1559.
[15]Zhao L,Li R,Gan YH.Knockdown of Yin Yang 1enhances anticancer effects of cisplatin through protein phosphatase2A-mediated T308dephosphorylation of AKT[J].Cell Death Dis,2018,9:747.
[16]Zhou L,Wang L,Lu L,et al.A novel target of microRNA-29,Ring1and YY1-binding protein(Rybp),negatively regulates skeletal myogenesis[J].J Biol Chem,2012,287:25255-25265.
[17]Shibue T,Weinberg RA.EMT,CSCs,and drug resistance:the mechanistic link and clinical implications[J].Nat Rev Clin Oncol,2017,14:611-629.
[18]Khachigian LM.The Yin and Yang of YY1in tumor growth and suppression[J].Int J Cancer.,2018,143:460-465.
[2]Weidle UH,Birzele F,Kollmorgen G,et al.Potential microRNA-related targets for therapeutic intervention with ovarian cancer metastasis[J].Cancer Genomics Proteomics,2018,15:1-15.
[3]Praestegaard C,Jensen A,Jensen SM,et al.Cigarette smoking is associated with adverse survival among women with ovarian cancer:Results from a pooled analysis of 19studies[J].Int J Cancer,2017,140:2422-2435.
[4]Bax HJ,Josephs DH,Pellizzari G,et al.Therapeutic targets and new directions for antibodies developed for ovarian cancer[J].MAbs,2016,8:1437-1455.
[5]黄艺,高月.MiR-223通过上皮间质转化抑制宫颈癌细胞侵袭转移[J].中国热带医学,2018,18:991-993,1003.
[6]丁婷,宫瑞婷,王晓强,等.YB-1对卵巢癌细胞EMT标志蛋白表达及黏附能力的影响研究[J].临床和实验医学杂志,2018,13:14-17.
[7]王冉冉,董珊珊,龙瀛,等.HMMR-AS1介导EMT在上皮性卵巢癌顺铂获得性耐药中的作用[J].肿瘤药学,2018,8:498-504.
[8]陈彦臻,刘沈林,邹玺.胃癌干细胞与上皮间质转化研究进展[J].中华肿瘤防治杂志,2018,25:750-754.
[9]Cho AA,Bonavida B.Targeting the overexpressed YY1in cancer inhibits EMT and metastasis[J].Crit Rev Oncog,2017,22:49-61.
[10]Sechler M,Parrish JK,Birks DK,et al.The histone demethylase KDM3A,and its downstream target MCAM,promote Ewing Sarcoma cell migration and metastasis[J].Oncogene,2017,36:4150-4160.
[11]Wang J,Wu X,Wei C,et al.YY1 Positively Regulates Transcription by Targeting Promoters and Super-Enhancers through the BAF Complex in Embryonic Stem Cells[J].Stem Cell Reports,2018,10:1324-1339.
[12]Huang T,Wang G,Yang L,et al.Transcription factor YY1modulates lung cancer progression by activating lncRNA-PVT1[J].DNA Cell Biol,2017,36:947-958.
[13]Balkhi MY,Wittmann G,Xiong F,et al.YY1 Upregulates checkpoint receptors and downregulates type I cytokines in exhausted,chronically stimulated human T cells[J].iScience,2018,27:105-122.
[14]Lu Z,Hong CC,Kong G,et al.Polycomb group protein YY1is an essential regulator of hematopoietic stem cell quiescence[J].Cell Rep,2018,22:1545-1559.
[15]Zhao L,Li R,Gan YH.Knockdown of Yin Yang 1enhances anticancer effects of cisplatin through protein phosphatase2A-mediated T308dephosphorylation of AKT[J].Cell Death Dis,2018,9:747.
[16]Zhou L,Wang L,Lu L,et al.A novel target of microRNA-29,Ring1and YY1-binding protein(Rybp),negatively regulates skeletal myogenesis[J].J Biol Chem,2012,287:25255-25265.
[17]Shibue T,Weinberg RA.EMT,CSCs,and drug resistance:the mechanistic link and clinical implications[J].Nat Rev Clin Oncol,2017,14:611-629.
[18]Khachigian LM.The Yin and Yang of YY1in tumor growth and suppression[J].Int J Cancer.,2018,143:460-465.