TRIB3通过激活β-catenin信号通路促进鼻咽癌细胞增殖和侵袭迁移

TRIB3 promotes the proliferation,invasion and migration of nasopharyngeal carcinoma cells through activatingβ-catenin signaling pathway

目的探讨TRIB3对鼻咽癌细胞增殖和侵袭迁移的影响,并研究其作用机制。方法使用荧光定量PCR检测鼻咽癌组织和正常癌旁组织中TRIB3的相对表达量,并检测正常鼻咽上皮细胞(N69)和鼻咽癌细胞(HNE-1、CNE-2Z、5-8F)中TRIB3的相对表达量。体外培养人鼻咽癌细胞HNE-1、CNE-2Z,每株细胞分2组:即阴性对照组(NC组)和敲低TRIB3组(sh-TRIB3组)。荧光定量PCR和Western blot法检测转染后各组细胞TRIB3的表达水平。敲低TRIB3后,CCK-8实验和集落克隆实验检测鼻咽癌细胞增殖能力,Transwell实验和划痕实验检测鼻咽癌细胞迁移和侵袭能力,Western blot法检测EMT相关蛋白Snail、E-cadherin、N-cadherin和Vimentin和β-catenin信号通路相关蛋白GSK-3β、p-GSK-3β、β-catenin和p-β-catenin的相对表达水平。结果荧光定量PCR结果显示,与癌旁组织相比,鼻咽癌组织中TRIB3表达高度上调(P<0.001);与正常鼻咽上皮细胞(N69)相比,鼻咽癌细胞(HNE-1、CNE-2Z)中TRIB3的表达显著升高(P<0.01)。在HNE-1和CNE-2Z细胞中,与NC组相比,sh-TRIB3组TRIB3的表达量下调(P<0.05),增殖、迁移和侵袭能力被显著抑制(P<0.05),EMT相关蛋白E-cadherin表达上调,Snail、N-cadherin和Vimentin表达下调(均P<0.05),β-catenin信号通路相关蛋白p-GSK-3β和β-catenin表达下调,GSK-3β和p-β-catenin表达上调(均P<0.05)。结论TRIB3可以通过激活β-catenin信号通路促进鼻咽癌细胞的增殖和侵袭迁移,提示TRIB3可能是鼻咽癌治疗的潜在靶点。

Objective To explore effects of TRIB3 on the proliferation,invasion and migration of nasopharyngeal carcinoma cells and its mechanism.Methods The relative expression of TRIB3 was detected in nasopharyngeal carcinoma tissues and normal paracancerous tissues,normal nasopharyngeal epithelial cells(N69)and nasopharyngeal carcinoma cells(HNE-1,CNE-2Z,5-8F)by fluorescence quantitative PCR.Human nasopharyngeal carcinoma cells HNE-1 and CNE-2Z were cultured in vitro,and were respectively divided into two groups:negative control group(NC group)and TRIB3 knockdown group(sh-TRIB3 group).The expression level of TRIB3 was detected by quantitative real-time polymerase chain reaction(RT-qPCR)and Western blot.After knockdown of TRIB3,the proliferation of nasopharyngeal carcinoma cells was detected by CCK-8 assay and colony cloning test,the migration and invasion abilities of nasopharyngeal carcinoma cells were detected by Transwell test and wound-healing test,and the relative expression levels of EMT-related proteins Snail,E-cadherin,N-cadherin,Vimentin andβ-catenin signaling pathway-related proteins GSK-3β,p-GSK-3β,β-catenin,p-β-catenin were detected by Western blot.Results RT-qPCR results showed that the expression of TRIB3 was significantly higher in nasopharyngeal carcinoma tissues than that in normal paracancerous tissues(P<0.001),and was also significantly higher in nasopharyngeal carcinoma cells(HNE-1,CNE-2Z)than that in normal nasopharyngeal epithelial cells(N69)(P<0.01).Compared with NC group,the expression of TRIB3 in HNE-1 and CNE-2Z cells was downregulated in sh-TRIB3 group(P<0.05),the proliferation,migration and invasion abilities were significantly inhibited(P<0.05),the expression of E-cadherin was upregulated,and the levels of snail,N-cadherin and Vimentin were downregulated(P<0.05),p-GSK-3βandβ-catenin were downregulated,and the expression of GSK-3βand p-β-catenin was upregulated(P<0.05).Conclusion TRIB3 can promote the proliferation,invasion and migration of nasopharyngeal carcinoma cells via the activation ofβ-catenin signaling pathway,suggesting that TRIB3 may be a potential target in the treatment of nasopharyngeal carcinoma.