miR-143-3p靶向调控HNRNPC抑制鼻咽癌的体外转移活性

miR-143-3p inhibits in vitro metastasis activity of nasopharyngeal carcinoma by targeting and regulating HNRNPC

[目的]探讨miR-143-3p与HNRNPC在鼻咽癌细胞中的相互作用关系。[方法]将鼻咽癌细胞分为4组:miR NC组、miR-143-3p mimic组、pcDNA3.1 NC组和pcDNA3.1 HNRNPC组。用结晶紫染色检测单克隆形成数目、流式细胞术检测细胞凋亡情况,同时RT-PCR和免疫组化实验测定miR-143-3p、HNRNPC的表达水平,蛋白免疫印迹法测定HNRNPC蛋白的表达水平。用生信学方法预测miR-143-3p与HNRNPC的结合位点,荧光素酶报告基因检测miR-143-3p对CNE2细胞HNRNPC活性的调控作用。[结果]miR-143-3p在鼻咽癌组织中的表达水平显著降低(1.25±0.03 vs 3.67±0.06),然而HNRNPC在鼻咽癌组织的表达水平升高(1.32±0.05 vs 0.46±0.03)。此外,miR-143-3p能够靶向抑制HNRNPC表达。在鼻咽癌细胞系中,miR-143-3p mimic处理完细胞之后,鼻咽癌细胞增殖能力受到抑制(58.15±7.28 vs 13.27±3.56),凋亡增加(9.05±0.28 vs 30.07±1.59)%。pcDNA3.1 HNRNPC处理完细胞之后,鼻咽癌细胞增殖能力提高(61.22±5.38 vs 113.57±5.87),凋亡减少(9.33±0.08 vs 5.01±0.09)%。[结论]miR-143-3p在鼻咽癌组织中低表达,miR-143-3p通过靶向作用HNRNPC进而抑制鼻咽癌细胞的生长,并促进鼻咽癌细胞凋亡。

[Objective]To investigate the interaction between miR-143-3p and HNRNPC in nasopharyngeal carcinoma cells.[Method]NPC cells were divided into four groups:miR NC group,miR-143-3p mimic group,pcDNA3.1 NC group and pcDNA3.1 HNRNPC group.The number of monoclonal colonies was detected by crystal violet staining,and cell apoptosis was measured by flow cytometry.The level of miR-143-3p and HNRNPC were determined by RT-PCR and immunohistochemistry,and the expression levels of HNRNPC proteins were determined by Western Blot.The binding sites of miR-143-3p and HNRNPC were predicted by bioinformatics methods,and the regulation effect of miR-143-3p on HNRNPC activity in CNE2 cells was detected by luciferase reporter gene.[Result]The expression level of miR-143-3p was significantly decreased in NPC tissues(1.25±0.03 vs 3.67±0.06),while the expression level of HNRNPC was increased in NPC tissues(1.32±0.05 vs 0.46±0.03).In addition,miR-143-3p could target and inhibit HNRNPC expression.After treatment with miR-143-3p mimic,the proliferation ability of NPC cells was inhibited(58.15±7.28 vs 13.27±3.56),and the apoptosis rate was increased(9.05±0.28 vs 30.07±1.59)%.After treatment with pcDNA 3.1 HNRNPC,the proliferation ability of NPC cells was increased(61.22±5.38 vs 113.57±5.87),and the apoptosis was decreased(9.33±0.08 vs 5.01±0.09)%.[Conclusion]miR-143-3p is lowly expressed in nasopharyngeal carcinoma tissues.miR-143-3p inhibits the growth of nasopharyngeal carcinoma cells and promotes the apoptosis of nasopharyngeal carcinoma cells by targeting HNRNPC.