目的探讨微小RNA-140-3p (miR-140-3p)对子宫肌瘤细胞增殖、迁移和侵袭的影响及作用机制。方法选取于2016年2月至2018年9月在西安市中医医院行手术治疗的59例子宫肌瘤患者为研究对象,同时期41例因子宫肌瘤切除全子宫的正常子宫肌组织作...目的探讨微小RNA-140-3p (miR-140-3p)对子宫肌瘤细胞增殖、迁移和侵袭的影响及作用机制。方法选取于2016年2月至2018年9月在西安市中医医院行手术治疗的59例子宫肌瘤患者为研究对象,同时期41例因子宫肌瘤切除全子宫的正常子宫肌组织作为对照,实时荧光定量PCR (RT-qPCR)检测组织中miR-140-3p和锚蛋白B (ANK2) mRNA表达水平。子宫肌瘤细胞分为miR-140-3p组(转染miR-140-3p模拟物mimics)、miR-NC组(转染mimics阴性对照序列)、si-ANK2组(转染ANK2小干扰RNA)、si-NC组(转染乱序无意义阴性序列)、miR-140-3p+pcDNA-ANK2组(共转染miR-140-3p mimcs与ANK2过表达载体)和miR-140-3p+pcDNA组(共转染miR-140-3p mimcs与空载体)。RT-qPCR和蛋白印迹(Western blot)分别检测miR-140-3p或ANK2蛋白水平验证转染效果,四甲基噻唑蓝染色法(MTT)检测各组细胞增殖情况,Transwell检测各组细胞的迁移和侵袭能力,Western blot检测各组细胞中细胞周期蛋白D1 (CyclinD1)、p21、基质金属蛋白酶2 (MMP-2)、E-钙黏附素(E-cadherin)蛋白表达。双荧光素酶报告基因实验验证子宫肌瘤细胞中miR-140-3p和ANK2调控关系。结果与正常宫颈组织比较,子宫肌瘤组织中miR-140-3p表达水平降低[(0.38±0.04) vs (1.06±0.09)],ANK2 mRNA表达水平升高[(2.61±0.24) vs (1.07±0.10)],差异均具有统计学意义(P<0.05);与miR-NC组比较,miR-140-3p组子宫肌瘤细胞存活率[(62.37±1.59)%vs (99.63±3.28)%]、迁移细胞数[(72.58±8.19) vs (139.25±12.34)]、侵袭细胞数[(53.91±6.18) vs (115.21±10.69)]、CyclinD1蛋白水平[(0.23±0.04) vs (0.98±0.11)]和MMP-2蛋白水平[(0.19±0.03) vs (0.53±0.06)]降低,而p21蛋白水平[(0.82±0.08) vs (0.29±0.05)]和E-cadherin蛋白水平[(0.57±0.07) vs (0.21±0.05)]升高,差异均具有统计学意义(P<0.05);与si-NC组比较,si-ANK2组子宫肌瘤细胞存活率[(72.59±2.08)%vs (98.67±3.58)%]、迁移细胞数[(75.69±8.54)vs (142.58±12.86)]、侵袭细胞数[(58.39±6.58) vs (123.67±展开更多
Background:Ovarian cancer is one of the most widespread malignant diseases of the female reproductive system worldwide.The plurality of ovarian cancer is diagnosed with metastasis in the abdominal cavity.Epithelial-me...Background:Ovarian cancer is one of the most widespread malignant diseases of the female reproductive system worldwide.The plurality of ovarian cancer is diagnosed with metastasis in the abdominal cavity.Epithelial-mesenchymal transition(EMT)exerts a vital role in tumor cell metastasis.However,it remains unclear whether long non-coding RNA(lncRNA)are implicated in EMT and influence ovarian cancer cell invasion and metastasis.This study was designed to investigate the impacts of lncRNA AC005224.4 on ovarian cancer.Methods:LncRNA AC005224.4,miR-140-3p,and snail family transcriptional repressor 2(SNAI2)expression levels in ovarian cancer and normal ovarian tissues were determined using real-time quantitative polymerase chain reaction(qRT-PCR).Cell Counting Kit-8(CCK-8)and Transwell(migration and invasion)assays were conducted to measure SKOV3 and CAOV-3 cell proliferation and metastasis.E-cadherin,N-cadherin,Snail,and Vimentin contents were detected using Western blot.Nude mouse xenograft assay was utilized to validate AC005224.4 effects in vivo.Dual-luciferase reporter gene assay confirmed the targeted relationship between miR-140-3p and AC005224.4 or SNAI2.Results:AC005224.4 and SNAI2 upregulation and miR-140-3p downregulation were observed in ovarian cancer tissues and cells.Silencing of AC005224.4 observably moderated SKOV3 and CAOV-3 cell proliferation,migration,invasion,and EMT process in vitro and impaired the tumorigenesis in vivo.miR-140-3p was a target of AC005224.4 and its reduced expression level was mediated by AC005224.4.miR-140-3p mimics decreased the proliferation,migration,and invasion of ovarian cancer cells.SNAI2 was identified as a novel target of miR-140-3p and its expression level was promoted by either AC005224.4 overexpression or miR-140-3p knockdown.Overexpression of SNAI2 also facilitated ovarian cancer cell viability and metastasis.Conclusion:AC005224.4 was confirmed as an oncogene via sponging miR-140-3p and promoted SNAI2 expression,contributing to better understanding of ovarian cancer 展开更多
文摘目的探讨微小RNA-140-3p (miR-140-3p)对子宫肌瘤细胞增殖、迁移和侵袭的影响及作用机制。方法选取于2016年2月至2018年9月在西安市中医医院行手术治疗的59例子宫肌瘤患者为研究对象,同时期41例因子宫肌瘤切除全子宫的正常子宫肌组织作为对照,实时荧光定量PCR (RT-qPCR)检测组织中miR-140-3p和锚蛋白B (ANK2) mRNA表达水平。子宫肌瘤细胞分为miR-140-3p组(转染miR-140-3p模拟物mimics)、miR-NC组(转染mimics阴性对照序列)、si-ANK2组(转染ANK2小干扰RNA)、si-NC组(转染乱序无意义阴性序列)、miR-140-3p+pcDNA-ANK2组(共转染miR-140-3p mimcs与ANK2过表达载体)和miR-140-3p+pcDNA组(共转染miR-140-3p mimcs与空载体)。RT-qPCR和蛋白印迹(Western blot)分别检测miR-140-3p或ANK2蛋白水平验证转染效果,四甲基噻唑蓝染色法(MTT)检测各组细胞增殖情况,Transwell检测各组细胞的迁移和侵袭能力,Western blot检测各组细胞中细胞周期蛋白D1 (CyclinD1)、p21、基质金属蛋白酶2 (MMP-2)、E-钙黏附素(E-cadherin)蛋白表达。双荧光素酶报告基因实验验证子宫肌瘤细胞中miR-140-3p和ANK2调控关系。结果与正常宫颈组织比较,子宫肌瘤组织中miR-140-3p表达水平降低[(0.38±0.04) vs (1.06±0.09)],ANK2 mRNA表达水平升高[(2.61±0.24) vs (1.07±0.10)],差异均具有统计学意义(P<0.05);与miR-NC组比较,miR-140-3p组子宫肌瘤细胞存活率[(62.37±1.59)%vs (99.63±3.28)%]、迁移细胞数[(72.58±8.19) vs (139.25±12.34)]、侵袭细胞数[(53.91±6.18) vs (115.21±10.69)]、CyclinD1蛋白水平[(0.23±0.04) vs (0.98±0.11)]和MMP-2蛋白水平[(0.19±0.03) vs (0.53±0.06)]降低,而p21蛋白水平[(0.82±0.08) vs (0.29±0.05)]和E-cadherin蛋白水平[(0.57±0.07) vs (0.21±0.05)]升高,差异均具有统计学意义(P<0.05);与si-NC组比较,si-ANK2组子宫肌瘤细胞存活率[(72.59±2.08)%vs (98.67±3.58)%]、迁移细胞数[(75.69±8.54)vs (142.58±12.86)]、侵袭细胞数[(58.39±6.58) vs (123.67±
基金Xinjiang Uygur Autonomous Region Science and Technology Supporting Xinjiang Project(2017E0263)Tianjin Science and Technology Support Program Project(18YFZCSY00100)+3 种基金Program for New Century Excellent Talents in University in China(NCET-11-1066)Training Plan of leading subject talents in Tianjin colleges and universitiesthe National Natural Science Foundation of China(81972572)CAMS Innovation Fund for Medical Sciences(2016-I2M-1-001)
文摘Background:Ovarian cancer is one of the most widespread malignant diseases of the female reproductive system worldwide.The plurality of ovarian cancer is diagnosed with metastasis in the abdominal cavity.Epithelial-mesenchymal transition(EMT)exerts a vital role in tumor cell metastasis.However,it remains unclear whether long non-coding RNA(lncRNA)are implicated in EMT and influence ovarian cancer cell invasion and metastasis.This study was designed to investigate the impacts of lncRNA AC005224.4 on ovarian cancer.Methods:LncRNA AC005224.4,miR-140-3p,and snail family transcriptional repressor 2(SNAI2)expression levels in ovarian cancer and normal ovarian tissues were determined using real-time quantitative polymerase chain reaction(qRT-PCR).Cell Counting Kit-8(CCK-8)and Transwell(migration and invasion)assays were conducted to measure SKOV3 and CAOV-3 cell proliferation and metastasis.E-cadherin,N-cadherin,Snail,and Vimentin contents were detected using Western blot.Nude mouse xenograft assay was utilized to validate AC005224.4 effects in vivo.Dual-luciferase reporter gene assay confirmed the targeted relationship between miR-140-3p and AC005224.4 or SNAI2.Results:AC005224.4 and SNAI2 upregulation and miR-140-3p downregulation were observed in ovarian cancer tissues and cells.Silencing of AC005224.4 observably moderated SKOV3 and CAOV-3 cell proliferation,migration,invasion,and EMT process in vitro and impaired the tumorigenesis in vivo.miR-140-3p was a target of AC005224.4 and its reduced expression level was mediated by AC005224.4.miR-140-3p mimics decreased the proliferation,migration,and invasion of ovarian cancer cells.SNAI2 was identified as a novel target of miR-140-3p and its expression level was promoted by either AC005224.4 overexpression or miR-140-3p knockdown.Overexpression of SNAI2 also facilitated ovarian cancer cell viability and metastasis.Conclusion:AC005224.4 was confirmed as an oncogene via sponging miR-140-3p and promoted SNAI2 expression,contributing to better understanding of ovarian cancer
