摘要
目的:探究miR-411-3p靶向p21活化激酶2(PAK2)基因介导Wnt信号通路对口腔鳞状细胞癌侵袭和转移的影响。方法:收集我院52例口腔癌组织及正常口腔黏膜上皮组织。免疫组化检测组织中PAK2阳性表达,qRT-PCR及Western blot检测miR-411-3p及PAK2表达。筛选PAK2高表达细胞系并将其分为:阴性对照组、miR-411-3p agomir组、miR-411-3p antagomir组、PAK2基因沉默组、联合组(miR-411-3p agomir联合PAK2沉默联合处理细胞)。qRT-PCR和Western blot检测各组细胞miR-411-3p、Wnt1、GSK3β、β-catenin、E-cadherin、N-cadherin表达,CCK8检测细胞活力,划痕实验、Transwell检测细胞迁移和侵袭。Annexin V-FITC/PI双染检测细胞凋亡情况。结果:与正常口腔黏膜上皮组织相比,癌组织中miR-411-3p表达下降,PAK2表达升高。与阴性对照组相比,miR-411-3p过表达或沉默PAK2能够抑制Wnt信号通路相关因子(Wnt1,β-catenin)及N-cadherin表达,促进GSK3β及E-cadherin表达,且联合组效果最为明显(P<0.05),抑制口腔鳞癌细胞活力,减少癌细胞迁移和侵袭,促进其凋亡,且联合组效果更明显(P<0.05)。结论:miR-411-3p可靶向抑制PAK2基因的表达介导Wnt信号通路进而抑制口腔鳞状细胞癌迁移、侵袭,促进癌细胞凋亡,有望成为口腔鳞状细胞癌治疗的重要靶点。
Objective:To explore effect of miR-411-3p on cell invasion and metastasis in oral squamous cell carcinoma by targeting p21 activated kinase 2(PAK2)via Wnt signaling pathway.Methods:Oral squamous cell carcinoma tissues and normal oral mucosal epithelial tissue,were collected from 52 patients in our hospital.Immunohistochemistry was adopted to detect positive expression of PAK2,qRT-PCR and Western blot were utilized to determine expressions of miR-411-3p and PAK2 in tissues.Oral squamous cell carcinoma cell line with highest expression of PAK2 was divided into Negative control group,miR-411-3p agomir group,miR-411-3p antagomir group,PAK2 shRNA group and combined treatment group(cells treated with miR-411-3p agomir and PAK2 silencing).qRT-PCR and Western blot were used to detect expressions of miR-411-3p,Wnt1,GSK3β,β-catenin,E-cadherin and N-cadherin in each group.CCK8 was used to detect cell viability.Wounding healing assay and Transwell assay were adopted to determine migration and invasion.Annexin V-FITC/PI double staining was used to detect cell apoptosis.Results:Compared with normal oral mucosal epithelial tissue,expression of miR-411-3p was decreased while PAK2 expression was increased in cancer tissue.Compared with Negative control group,miR-411-3p overexpression or PAK2 silencing could inhibit expressions of Wnt signaling pathway related factors(Wnt1,β-catenin)and N-cadherin,induce expressions of GSK3βand E-cadherin,combined treatment group showed most obvious change(P<0.05),and inhibited cell viability of oral squamous cell carcinoma cells,downregulated migration and invasion of cancer cells,induced apoptosis while combined treatment group showed more profound change(P<0.05).Conclusion:miR-411-3p could inhibit migration and invasion of oral squamous cell carcinoma cells,induce cell apoptosis through targeting PAK2 via Wnt signaling pathway,who has potential to become an important therapeutic target in oral squamous cell carcinoma.
作者
麦明思
赵焱
马锴
MAI Ming-Si;ZHAO Yan;MA Kai(Department of Stomatology,Sanya People′s Hospital,Sanya 572000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第17期2109-2114,共6页
Chinese Journal of Immunology
基金
海南省自然科学基金(814390)资助。
