MRTFA在卵巢癌组织中的表达及对细胞增殖、迁移和侵袭能力的影响

Expression of MRTFA in ovarian cancer tissue and its effect on cell proliferation,migration and invasion

目的探讨心肌素相关转录因子A(MRTFA)在卵巢癌组织中的表达及对细胞增殖、迁移和侵袭能力的影响。方法选取82例卵巢癌患者,应用实时荧光定量聚合酶链式反应(PCR)检测卵巢癌组织和癌旁组织中MRTFA mRNA的相对表达量,分析不同临床特征的卵巢癌患者卵巢癌组织中MRTFA mRNA的相对表达量。培养卵巢癌SKOV3细胞,并分为MRTFA干扰组、阴性对照组和对照组,噻唑蓝(MTT)法检测细胞的增殖活性,划痕实验检测细胞的迁移能力,Transwell小室检测细胞的侵袭能力,蛋白质印迹(Western blot)法检测细胞中MRTFA、血清应答因子(SRF)、E-钙黏素(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)的相对表达量。结果卵巢癌组织中MRTFA mRNA的相对表达量为(1.86±0.12),明显高于癌旁组织的(1.15±0.10),差异有统计学意义(P﹤0.01)。不同肿瘤直径、国际妇产科联盟(FIGO)分期、分化程度和淋巴结转移情况的卵巢癌患者卵巢癌组织中MRTFA mRNA的相对表达量比较,差异均有统计学意义(P﹤0.05)。培养24、48、72、96 h后,MRTFA干扰组细胞的吸光度值均低于阴性对照组和对照组(P﹤0.05);培养24、48 h后,MRTFA干扰组细胞的划痕愈合率均低于对照组和阴性对照组(P﹤0.05);培养24 h后,MRTFA干扰组的侵袭细胞数少于对照组和阴性对照组(P﹤0.05)。MRTFA干扰组细胞中MRTFA、SRF和α-SMA蛋白的相对表达量均低于阴性对照组和对照组,E-cadherin蛋白的相对表达量高于阴性对照组和对照组(P﹤0.05)。结论卵巢癌组织中MRTFA蛋白呈高表达,特异性抑制MRTFA基因表达可抑制细胞的增殖、迁移和侵袭能力,其机制可能与抑制SRF蛋白介导的上皮-间充质转化过程有关。

Objective To investigate the expression of myocardin related transcription factor A(MRTFA)in ovarian cancer tissue,and explore its effect on cell proliferation,migration and invasion ability.Method The research included 82 cases of patients with ovarian cancer.The relative expressions of MRTFA mRNA in ovarian cancer tissues and adjacent tissues were detected by real-time fluorescence quantitative polymerase chain reaction(PCR),and the different expression level of MRTFA mRNA in ovarian tissues with varying clinical features were investigated.Ovarian cancer SKOV3 cells were cultured and divided into MRTFA interference group,negative control group and control group.Cell proliferation activity was detected by thiazolyl blue tetrazolium bromide(MTT)assay.Cell migration was assessed by scratch test.Cell invasion was determined using Transwell assay.Besides,The expressions of MRTFA,serum response factor(SRF),E-cadherin and α-smooth muscle actin(α-SMA)proteins were detected by Western blot.Result The relative expression level of MRTFA mRNA in ovarian cancer tissues was(1.86±0.12),which was significantly higher than that in adjacent tissues at(1.15±0.10),the difference was statistically significant(P<0.01).The relative expression level of MRTFA mRNA in ovarian cancer tissues was related to tumor diameter,International Federation of Gynecology and Obstetrics(FIGO)stage,degree of differentiation and presence of lymph node metastasis(P<0.05).The absorbance values at 24,48,72,96 h in the MRTFA interference group were lower than those in the negative control group and control group(P<0.05).The scratch wound healing rates after culturing for 24 h and 48 h and the number of invasive cells in 24 h of culturing in the MRTFA interference group were lower than those in the negative control group and control group(P<0.05).The relative expression levels of MRTFA,SRF,and α-SMA proteins in the MRTFA interference group were lower than those in the negative control group and the control group,the relative expression level of E-cadherin in the MRTFA interference group were higher than that in the negative control group and the control group(P<0.05).Conclusion MRTFA protein was highly expressed in ovarian cancer tissues.Specific inhibition of MRTFA gene expression could restrain the cell proliferation,migration and invasion abilities,and its mechanism might be related to inhibition of SRF protein-mediated epithelial-mesenchymal transition process.