miR-454-3p通过靶向STAT3调控CNE-2人鼻咽癌细胞上皮-间质转化

The miR-454-3p regulates epithelial-mesenchymal transition in CNE-2 human nasopharyngeal carcinoma cells by targeting STAT3

目的:探讨miR-454-3p和STAT3在鼻咽癌组织中的表达情况,以及miR-454-3p靶向STAT3对鼻咽癌细胞上皮-间质转化的影响。方法:收集2023年6月~8月右江民族医学院附属医院和百色市人民医院的鼻咽癌组织和正常组织各5例,采用qRT-PCR检测各样本中miR-454-3p与STAT3的表达情况;以未处理的鼻咽癌细胞CNE-2作为空白对照组(blank group),采用LipofectamineTM3000将miR-454-3p mimics、mimics NC(negative control)、miR-454-3p inhibitor、inhibitor NC(negative control)分别转染CNE-2细胞作为miR-454-3p mimics组、mimics NC组、miR-454-3p inhibitor组、inhibitor NC组;利用Targetscan预测miR-454-3p与STAT3的靶向关系并用双荧光素酶实验验证;分别采用qRT-PCR检测转染效率及转染后STAT3的mRNA水平变化;划痕愈合实验、Transwell小室实验检测细胞的迁移和侵袭能力;Western blot检测STAT3和上皮-间质转化相关蛋白的表达。结果:鼻咽癌组织的miR-454-3p表达量较正常组织降低,而STAT3表达量升高(P<0.001);双荧光素酶实验提示miR-454-3p靶向调控STAT3表达(P<0.001);与空白组和mimics NC组相比,miR-454-3p mimics组的STAT3、N-cadherin、Vimentin蛋白表达下调,Ecadherin蛋白表达上调,细胞的侵袭和迁移能力均减弱(P<0.05);与空白组和inhibitor NC组相比,miR-454-3p inhibitor组的STAT3、N-cadherin、Vimentin蛋白表达上调,E-cadherin蛋白表达下调,细胞的侵袭和迁移能力均增强(P<0.05)。结论:在鼻咽癌组织中miR-454-3p表达下调,而STAT3表达上调,在CNE-2人鼻咽癌细胞中miR-454-3p可以显著降低STAT3的活性,有效抑制细胞的侵袭和迁移。

Objective:To explore the levels of miR-454-3p and STAT3 in nasopharyngeal carcinoma(NPC)tissues,as well as the effect of miR-454-3p targeting STAT3 on the process of epithelial-mesenchymal transition(EMT)in NPC cells.Methods:NPC tissue and healthy tissue were collected in five cases from the Affiliated Hospital of Youjiang Medical University for Nationalities and Baise People's Hospital from June to August 2023.qRT-PCR detected the expression of miR-454-3p and STAT3 in each tissue sample.Untreated NPC cells CNE-2 was used as a blank control group(Blank group).miR-454-3p mimics,mimics NC(negative control),miR-454-3p inhibitor and inhibitor NC(negative control)were transfected into CNE-2 cells by LipofectamineTM 3000 as miR-454-3p mimics group,mimics NC group,miR-454-3p inhibitor group and inhibitor NC group.The targeting relationship of miR-454-3p and STAT3 was predicted by Targetscan and confirmed by the dual-luciferase assay.The transfection efficiency and STAT3 mRNA levels were assessed using qRT-PCR following transfection.Wound healing and transwell experiments were executed to assess cell migratory and invasive abilities.The expression levels of STAT3 and proteins associated with EMT were assessed using Western blot analysis.Results:The expression of miR-454-3p was reduced in NPC tissues relative to healthy tissues,while the expression of STAT3 was elevated(all P<0.001).The dual-luciferase assay findings indicated that miR-454-3p targeting regulates expression of STAT3(P<0.001).Compared to both the blank group and mimics NC group,STAT3,N-cadherin,and Vimentin expressions of the miR-454-3p mimics group were reduced.Conversely,it exhibited an increase in E-cadherin expression.The migratory and invasive capabilities of the cells were weakened(all P<0.05).Compared to the blank group and inhibitor NC group,the expressions of STAT3,N-cadherin,and Vimentin in the miR-454-3p inhibitor group were upregulated.In contrast,the expression of E-cadherin was downregulated.Cells showed both enhanced migratory and invasive capacity(all P<0.05).Conclusion:In NPC tissues,miR-454-3p demonstrated downregulation,whereas STAT3 exhibited upregulation.Notably,within CNE-2 human NPC cells,miR-454-3p significantly reduced STAT3 activity and effectively suppressed cell invasion and migration.