MiRNA-206通过抑制CDK9表达对鼻咽癌CNE2细胞的生长及转移的影响

Effect of miRNA-206 on the growth and metastasis of nasopharyngeal carcinoma CNE2 cells by inhibiting expression of CDK9

目的:探讨miRNA-206对人鼻咽癌CNE2细胞的增殖和转移能力的影响,并分析细胞周期蛋白依赖性激酶9(cyclin-dependent kinase 9,CDK9)在其中的作用。方法:采用实时荧光定量PCR(real-timequantitative polymerase chain reaction,RT-qPCR)法检测miR-206在鼻咽上皮NP69细胞和人鼻咽癌CNE2细胞中的表达水平后,将miR-206模拟物或对照物分别导入CNE2细胞。通过CCK-8试验、Annexin V-FITC/PI染色、克隆形成试验、Transwell细胞迁移试验和Transwell细胞侵袭试验分别评价miR-206过表达对CNE2细胞活力、凋亡、克隆形成、迁移和侵袭的影响。最后采用蛋白质印迹法检测CDK9在NP69细胞和CNE2细胞中的表达。进一步检测过表达CDK9对miR-206过表达CNE2细胞的迁移和侵袭的影响。结果:MiR-206在CNE2细胞系中低表达。MiR-206过表达降低CNE2细胞的活性、克隆形成、迁移、侵袭并诱导细胞凋亡。CNE2细胞中CDK9呈现高表达。过表达miR-206显著抑制CNE2细胞中CDK9表达水平;而CDK9过表达能逆转miR-206过表达对CNE2细胞侵袭与迁移的抑制作用。结论:MiRNA-206对CNE2细胞的生长和转移起抑制作用,可通过抑制CDK9表达来介导。

Objective: To investigate the effect of miR-206 on proliferation and metastasis of human nasopharyngeal carcinoma CNE2 cells, and to analyze the role of cyclin-dependent kinase 9 (CDK9). Methods: Expression levels of miR-206 in nasopharyngeal epithelial NP69 cells and human nasopharyngeal carcinoma CNE2 cells were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The mimic and control of miR- 206 were introduced into CNE2 cells, respectively. CCK-8 assay, Annexin V-FITC/PIstaining, clone formation assay, transwell migration assay and transwell invasion assay were used to evaluate the effect of miR-206 over expression on cell viability, apoptosis, colony formation, migration and invasion in CNE2 cells, respectively. Finally, Expression levels of CDK9 in CNE2 cells and NP69 cells were detected by Western blot. The effect overexpression of CDK9 on migration and invasion in over-expression of miR-206 of CNE2 cells was further detected. Results: MiR-206 was lower expression in CNE2 cells. Over-expression of miR-206 decreased cell viability, colony formation, migration and invasion and induced apoptosis in CNE2 cells. CDK9 was higher expression in CNE2 cells. Over-expression of miR-206 inhibited Expression of CDK9 in CNE2 cells. However, over-expression of CDK9 could reverse the inhibitory effect of over-expression of miR-206 on the invasion and migration of CNE2 cells. Conclusion: MiRNA-206 inhibits the growth and metastasis in CNE2 cells, which may be mediated though inhibiting Expression of CDK9.