摘要
目的 探讨长链非编码RNA UPK1A反义RNA 1(UPK1A-AS1)在子宫内膜癌(EC)中的表达及其调控EC发生的分子机制.方法 利用starBase数据库或qPCR法检测UPK1A-AS1 在EC中的表达.将Ishikawa细胞分为Control组(空白)、si-NC组(阴性对照)、si-UPK1A-AS1 组(下调UPK1A-AS1 表达)和si-UPK1A-AS1+Wnt组(下调UPK1A-AS1 表达联合激活Wnt信号通路).通过细胞计数试剂盒-8(CCK-8)、划痕实验和Transwell实验评估Ishikawa细胞的增殖、迁移和侵袭;qPCR和Western blot检测Wnt1、β-catenin和c-Myc的表达.结果 UPK1A-AS1 在EC组织中的表达异常上调.与hEEC细胞相比,UPK1A-AS1 在RL-952、KLE、HEC-1B和Ishikawa细胞中高表达.与si-NC组比较,si-UPK1A-AS1 组的细胞活力、划痕愈合率和侵袭细胞数均降低(P＜0.01).与si-UPK1A-AS1 组相比,si-UPK1A-AS1+Wnt组的细胞活力、划痕愈合率和侵袭细胞数均增加(P＜0.01).si-UPK1A-AS1 组Wnt1、β-catenin、c-Myc的mRNA和蛋白水平均低于si-NC组(P＜0.01).si-UPK1A-AS1+Wnt组Wnt1、β-catenin、c-Myc的表达均高于si-UPK1A-AS1 组(P＜0.01).结论 UPK1A-AS1 激活Wnt信号通路促进了EC的进展和转移.UPK1A-AS1/Wnt/β-catenin/c-Myc轴可能是EC的潜在治疗靶点.
Abstract
Objective To investigate the expression level of long non-coding RNA UPK1A antisense RNA 1(UPK1A-AS1)in endometrial carcinoma(EC)and the molecular mechanism that regulates the development of EC.Methods Expressions of UPK1A-AS1 in EC were detected in starBase database or by qPCR.Ishikawa cells were allocated into Control group(blank),si-NC group(negative control),si-UPK1A-AS1 group(down-regulation of UPK1A-AS1)and si-UPK1A-AS1+Wnt1 group(down-regulation of UPK1A-AS1 plus activation of Wnt signaling pathway).Cell Counting Kit-8(CCK-8),Wound-healing and Transwell assays were conducted to assess the proliferation,migration,and invasion of Ishikawa cells.Moreover,qPCR and Western blot were carried out to determine the expressions of Wnt1,β-catenin and c-Myc.Results The expressions of UPK1A-AS1 were found to be aberrantly upregulated in EC tissues.Compared with the hEEC cells,UPK1A-AS1 were high-expressed in RL-952,KLE,HEC-1B and Ishikawa cells(P＜0.01).Compared with si-NC group,the cell viability,healing rate and number of invaded cells in si-UPK1A-AS1 group were decreased(P＜0.01).While the cell viability,healing rate and number of invaded cells in si-UPK1A-AS1+Wnt1 group were increased as compared to si-UPK1A-AS1 group(P＜0.01).Besides,si-UPK1A-AS1 group showed lower expressions of Wnt1,β-catenin and c-Myc in both mRNA and protein levels than si-NC group(P＜0.01),but si-UPK1A-AS1+Wnt1 group exhibited higher expressions of Wnt1,β-catenin and c-Myc as compared with si-UPK1A-AS1 group(P＜0.01).Conclusion UPK1A-AS1 activates Wnt signaling to promote the progression and metastasis of EC.The UPK1A-AS1/Wnt/β-catenin/c-Myc axis could be a potential therapeutic target of EC.
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