microRNA-622调控DYRK2在结肠癌中表达并促进结肠癌细胞SW1116侵袭转移

目的 探讨微小RNA622(microRNA-622,miR-622)及双重特异性酪氨酸调节激酶2(DYRK2)在结肠癌组织及结肠细胞系SW1116、SW480中的表达情况并研究其对SW1116侵袭转移能力的影响.方法 选取82例结肠癌及癌旁组织标本,培养结肠癌细胞系SW1116、SW480及正常结肠上皮细胞系NCM460细胞.Real time PCR检测组织及细胞中miR-622的表达,Real time PCR、免疫组织化学、Western blot检测DYRK2基因及蛋白的表达并行Pearson相关性分析.在SW1116中转染miR-622 mimics上调miR-622表达,同时对照(NC)组转染阴性序列并验证,Real time PCR及Western blot进一步检测上调miR-622后SW1116中DYRK2基因及蛋白表达水平,同时用Transwell法检测SW1116细胞侵袭转移能力的变化.结果 Real time PCR及Western blot结果显示,相比于癌旁组织和正常结肠上皮细胞系NCM460,结肠癌组织及结肠癌细胞SW1116中miR-622 mRNA呈高表达而DYRK2 mRNA及蛋白呈低表达,两者表达呈明显负相关(r=0.916,P＜0.01).转染miR-622 mimics后,Real time PCR及Western blot结果显示,相比于NC组,miR-622 mimics组DYRK2 mRNA及蛋白表达水平减低(P＜0.01).相应的,Transwell结果显示,相比于NC组,SW1116细胞转染miR-622 mimics后侵袭转移能力明显增强(P＜0.01).结论 结肠癌中miR-622呈高表达而DYRK2呈低表达,上调miR-622可负性调控DYRK2表达并促进SW1116细胞侵袭转移.

MicroRNA-622 regulates DYRK2 expression in colon cancer and promotes migration in colon cancer cell SW1116

Objective To investigate the expressiorn of microRNA-622(miR-622) and dual specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2) in colon cancer tissues and cell lines and explore the effect of miR-622 on SW11l6 cells migration and invasion.Methods Eighty-two colon cancer tissues and paired para-tumor tissue specimens were collected.C.olon cancer cell line SW1116,SW480 and normal human colon epithelial cell line NCM460 were cultured.MiR-622 was detected by using Real time PCR,DYRK2 expression was measured by using immunohistochemistry,Real time PCR anid Western blot in tissue level and cell level,respectively.The relation of miR-622 and DYRK2 was analyzed by Pearson correlation analysis.miR-622 mimics transfection was conducted to up-regulate miR-622,while negative control,NC group were transfected with control sequence.Expression of DYRK2 was evaluated by using Real time PCR and Western blot,while Transwell chamber assays were used to assess the migration ability changes.Results Real time PCR and Western blot results showed that miR-622 mRNA was highly expressed in colorectal cancer tissue and colon cancer cell SW1116,whereas DYRK2 mRNA and protein were lowly expressed when compared with paracancerous tissue and normal colonic epithelial cell line NCM460.An obvious negative correlation was showed between miR-622 and DYRK2(r=0.916,P＜0.01).Compared to NC group,DYRK2 mRNA and protein expression were down-regulated after transfection of miR-622 mimics,which was observerd through Real time PCR and Western blot(P＜0.01).Correspondingly,compared to NC group,the migration ability of SW116 was remarkably enhanced after transfection of miR-622 mimics(P＜0.01).Conclusion The expression of miR-622 is high and DYRK2 is low in colon cancer.Up-regulation of miR-622 could negatively regulate DYRK2 expression and promote SW1116 cells migration.