TRIM59对人鼻咽癌细胞增殖、凋亡及EMT的影响

Effects of TRIM59 on proliferation,apoptosis and EMT of human nasopharyngeal carcinoma cells

目的探讨三基序蛋白59(tripartite motif containing 59,TRIM59)对人鼻咽癌细胞增殖、凋亡、上皮间质转化(epithelial-mesenchymal transition,EMT)的影响及相关分子机制。方法Western blot检测鼻咽癌细胞系HNE1、CNE-2Z和人正常鼻黏膜上皮细胞系(HNEpC)中TRIM59的蛋白表达情况。实验中HNE1和CNE-2Z分别构建si-NC组(Lip2000+si-NC)、si-TRIM59组(Lip2000+si-TRIM59)和mimic-NC组(Lip2000+mimic-NC)、TRIM59-mimics组(Lip2000+mimic-TRIM59)。CCK-8和集落形成实验检测细胞增殖情况;流式细胞术检测TRIM59对鼻咽癌细胞凋亡的影响;细胞划痕试验和Transwell检测细胞上皮-间质转化(EMT)能力;同时通过Western blot分析下调或上调TRIM59后鼻咽癌细胞中凋亡相关蛋白(Bcl-2和Bax)的表达情况;通过Western blot分析TRIM59对鼻咽癌细胞中EMT相关蛋白(E-cadherin、N-cadherin、Vimentin)的影响。结果与人正常鼻黏膜上皮细胞系(HNEpC)相比,TRIM59在HNE1和CNE-2Z中表达上调(P<0.05)。与si-NC组相比,si-TRIM59组细胞中TRIM59的蛋白表达量明显下降(P<0.05);与mimic-NC组相比,TRIM59-mimics组细胞中TRIM59的蛋白表达量明显上升(P<0.05)。与si-NC组相比,si-TRIM59组细胞的增殖受到抑制(P<0.05);与mimic-NC组相比,TRIM59-mimics组细胞增殖明显升高(P<0.05)。与si-NC组相比,si-TRIM59组细胞的凋亡数量明显上升,Bax上升,Bcl-2下降(均P<0.05);与mimic-NC组相比,TRIM59-mimics组细胞的凋亡数量明显下降,Bax下降,Bcl-2上升(均P<0.05)。与si-NC组相比,si-TRIM59组细胞的EMT能力明显下降,E-cadherin表达上升,N-cadherin和Vimentin表达下降(均P<0.05);与mimic-NC组相比,TRIM59-mimics组细胞的EMT能力明显上升,E-cadherin表达下降,N-cadherin和Vimentin表达上升(均P<0.05)。结论下调TRIM59抑制鼻咽癌细胞的增殖及EMT过程,促进凋亡,上调TRIM59可促进鼻咽癌细胞的增殖及EMT过程,抑制凋亡。

Objective To investigate the effects of TRIM59 on the proliferation,apoptosis and EMT of human nasopharyngeal carcinoma cells and its related molecular mechanisms.Methods Western blot was used to detect the protein expression of TRIM59 in nasopharyngeal carcinoma cell lines HNE1,CNE-2Z and human normal nasal epithelial cell line(HNEpC).In the experiment,HNE1 and CNE-2Z were respectively divided into si-NC group(Lip2000+si-NC),si-TRIM59 group(Lip2000+si-TRIM59),mimic-NC group(Lip2000+mimic-NC),and TRIM59-mimics group(Lip2000+mimic-TRIM59).CCK-8 and colony formation assay were used to detect cell proliferation,and flow cytometry was used to detect the effect of TRIM59 on apoptosis of nasopharyngeal carcinoma cells.Cell scratch test and Transwell were used to detect the ability of EMT.At the same time,the expression of apoptosis-related proteins(Bcl-2 and Bax)was detected in nasopharyngeal carcinoma cells after down-regulation or up-regulation of TRIM59 by Western blot.The effect of TRIM59 on EMT-related proteins(E-cadherin,N-cadherin,Vimentin)in nasopharyngeal carcinoma cells was analyzed by Western blot.Results Compared with HNEpC,the expression of TRIM59 was up-regulated in HNE1 and CNE-2Z(P<0.05).Compared with si-NC group,the protein expression of TRIM59 in si-TRIM59 group was significantly decreased(P<0.05).Compared with mimic-NC group,the protein expression of TRIM59 in TRIM59-mimics group was significantly increased(P<0.05).Compared with si-NC group,the cell proliferation was inhibited in si-TRIM59 group(P<0.05);compared with mimic-NC group,the cell prolife-ration promoted in TRIM59-mimics group(P<0.05).Compared with si-NC group,the number of apoptotic cells in si-TRIM59 group was increased significantly,Bax was increased,and Bcl-2 was decreased(P<0.05).Compared with mimic-NC group,the number of apoptotic cells in TRIM59-mimics group was decreased significantly,Bax was decreased,and Bcl-2 was increased(P<0.05).Compared with si-NC group,the EMT ability of cells in si-TRIM59 group was significantly decreased,the expression of E-cadherin was increased,and the expression levels of N-cadherin and Vimentin were decreased(P<0.05).Compared with mimic-NC group,the EMT ability was significantly increased in TRIM59-mimics group,the expression of E-cadherin was decreased,and the expression of N-cadherin and Vimentin was increased(P<0.05).Conclusion Down-regulation of TRIM59 inhibits the proliferation and EMT process of nasopharyngeal carcinoma cells and promotes the apoptosis.Up-regulation of TRIM59 promotes the proliferation and EMT process of nasopharyngeal carcinoma cells and inhibits the apoptosis.