Abstract：Objective To investigate the effect of miR-582-5p on the proliferation, invasion, and migration of cisplatin-resistant nasopharyngeal carcinoma cells and its mechanism. Methods Nasopharyngeal carcinoma HNE1/DDP cells were cultured and divided into control group and miR-582-5p group. The control group was transfected with miR-582-5p negative control reagent, whereas the miR-582-5p group was transfected with miR-582-5p mimics. MiR-582-5p and serine-threonine kinase 3(AKT3) mRNA expression was detected by quantitative real-time polymerase chain reaction(qRT-PCR), cell viability was detected by cell counting kit(CCK-8) assay, cell proliferation was detected by colony formation assay, and cell motility was detected by wound healing assay. The invasion and migration abilities of the cells were detected by Transwell chamber method, AKT3 protein expression was detected by Western blot, and the targeting relationship between miR-582-5p and AKT3 was detected by luciferase reporter experiment. Results (1) The relative mRNA expression levels of miR-582-5p in the control and miR-582-5p groups detected by qRT-PCR were 1.02±0.08 and 3.01±0.05, respectively. Compared with the control group, the relative mRNA expression level of miR-582-5P in the miR-582-5P group increased, and the difference was statistically significant (t=38.76, P<0.001). (2) The results of cell viability test showed that compared with the control group, the optical density (OD) of cell culture in the miR-582-5p group at 0 h was not statistically different (P=1.000). The OD values at 24, 48, and 72h decreased, and the differences were statistically significant (all P values <0.001). (3) The results of cell proliferation test showed that the colony number of HNE1/DDP cells in the control and miR-582-5p groups were 362.3±13.1 and 155.7±5.0, respectively; the colony number of HNE1/DDP cells in the miR-582-5p group was significantly lower than that in the control group (t=25.59, P<0.001). (4) The detection results of cell movement, invasion, and migration showed that the cell migration rate, cell invasion number, and cell migration number in the miR-582-5p group (62.0%±5.0%, 804.3±3.8, 631.0±1.0, respectively) were lower than those in the control group (29.3%±4.0%, 434.0±6.1, 373.3±7.6, respectively), and the differences were statistically significant (t=8.80, 89.53, 58.43; all P values <0.001). (5) The relative expression levels of AKT3 mRNA and protein in the miR-582-5p group (0.99±0.07, 1.34±0.02, respectively) were significantly lower than those in the control group (0.59±0.04, 0.60±0.03, respectively) (t=8.64, 32.03; all P values <0.001). Conclusion MiR-582-5p upregulation can inhibit the proliferation, invasion, and migration of cisplatin-resistant nasopharyngeal carcinoma cells, which may be related to the downregulation of AKT3 expression.
刘耘帆, 马俊结, 苏凯, 王晓敏, 李慧, 马士崟. miR-582-5p对顺铂耐药鼻咽癌细胞增殖、侵袭与迁移的影响及其机制[J]. 中华解剖与临床杂志, 2023, 28(6): 400-405.
Liu Yunfan, Ma Junjie, Su Kai, Wang Xiaomin, Li Hui, Ma Shiyin. Effect and mechanism of miR-582-5p on proliferation, invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells. Chinese Journal of Anatomy and Clinics, 2023, 28(6): 400-405.
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2023, Vol. 28 
