UNR调控上皮性卵巢癌细胞上皮间质转化的作用

摘要

目的 检测UNR基因在上皮性卵巢癌细胞中的表达,并观察其对卵巢癌细胞上皮间质转化(EMT)的影响.方法 采用实时荧光定量PCR(qPCR)检测并比较体外培养的人上皮性卵巢癌细胞系SKOV-3、HO-8910、ES-2和永生化人卵巢上皮细胞系MOODY中UNR基因的表达,采用qPCR和Western blot检测卵巢癌SKOV-3细胞的UNR-RNAi干扰效率,应用Transwell侵袭小室实验检测SKOV-3细胞的侵袭能力,采用Western blot检测SKOV-3细胞E-cadherin的蛋白表达.结果 UNR在上皮性卵巢癌细胞系SKOV-3、HO-8910、ES-2中均有显著表达.shUNR转染SKOV-3后,Transwell侵袭小室实验显示,UNR-RNAi组穿膜细胞数为每视野(29.33±5.14)个,显著低于阴性对照组(76.00±5.42)个(P＜0.01).此外,Western blot显示UNR基因干扰后SKOV-3细胞中E-eadherin的蛋白表达显著上调.结论 shUNR可以抑制卵巢癌SKOV-3细胞UNR基因的表达,降低卵巢癌细胞的侵袭能力.干扰后E-cadherin的表达上调提示UNR可能促进卵巢癌EMT的发生,以UNR基因为靶点的干扰技术有望成为卵巢癌治疗的新策略.%Objective To detect the expression of UNR gene in epithelial ovarian cancer cells,and to observe the effect of UNR gene on epithelial mesenchymal transition (EMT) of ovarian cancer cells.Methods Real-time quantitative PCR (qPCR) was used to detect and compare the expression of UNR gene in human epithelial ovarian cancer cell lines SKOV-3,HO-8910,ES-2 and immortalized human ovarian epithelial cell lines MOODY in vitro,qPCR and Western blot were used to detect the UNR-RNAi interfering efficiency of ovarian cancer cell SKOV-3,the invasion ability of SKOV-3 cells was detected by Transwell invasion assay.The protein expression of E-cadherin in SKOV-3 cells was detected by Western blot.Results UNR was significantly expressed in the epithelial ovarian cancer cell lines SKOV-3,HO-8910 and ES-2.After SKOV-3 was transfected by shUNR,Transwell assay showed that the number of transmembrane cells in UNR-RNAi group was (29.33±5.14) per field,which was significantly lower than that in negative control group [(76.00±5.42) per field] (P ＜ 0.01).In addition,Western blot showed that the expression of E-cadherin protein in SKOV-3 cells was significantly up-regulated after UNR gene interference.Conclusion shUNR can markedly inhibit the expression of UNR gene in ovarian cancer SKOV-3 cells,decrease the invasive ability of ovarian cancer cells.The up-regulated expression of E-cadherin after interference shows that UNR may promote the occurrence of ovarian cancer EMT,thus,the interfering technology taking UNR gene as target is expected to become a new strategy for the treatment of ovarian cancer.