摘要
目的 探讨微小RNA-572(miR-572)靶向调控蛋白磷酸酶2调节亚基2C(PPP2R2C)对鼻咽癌细胞增殖的影响及其机制。方法 CNE细胞分为对照组、NC组(转染negative control)、PPP2R2C组(转染PPP2R2C)、miR-in组(转染miR-572 inhibitor)和miR-in/siPPP2R2C组(转染miR-572 inhibitor和PPP2R2C-siRNA)。以细胞计数试剂盒-8(CCK-8)检测细胞增殖能力,以克隆形成实验检测细胞集落形成能力,以流式细胞仪检测不同细胞周期的细胞百分比,以蛋白质印迹(Western blot)法检测PPP2R2C蛋白表达水平,通过Targetscan在线分析数据库和双荧光素酶报告基因分别预测和验证miR-572和PPP2R2C之间的靶向调控关系。结果 对照组、NC组、PPP2R2C组、miR-in组和miR-in/siPPP2R2C组细胞增殖活力分别为(94.68±4.97)%、(93.85±3.88)%、(67.63±5.95)%、(52.37±5.60)%和(95.85±2.54)%,细胞克隆形成率分别为(81.32±2.56)%、(80.82±4.32)%、(57.63±5.95)%、(51.54±3.48)%和(79.47±4.37)%,细胞周期G0/G1期细胞比例分别为(40.59±1.84)%、(41.06±1.94)%、(59.36±2.02)%、(63.47±2.24)%和(39.32±1.01)%,S期比例分别为(38.30±1.01)%、(36.25±1.08)%、(20.69±1.15)%、(16.01±1.07)%和(39.71±1.50)%。与对照组、NC组比较,PPP2R2C组、miR-in组细胞增殖活力、细胞克隆形成率和S期细胞比例显著降低,G0/G1期细胞比例显著升高,差异均有统计学意义(均P<0.01),miR-in/siPPP2R2C组上述指标差异均无统计学意义(均P>0.05)。miR-in组和NC组PPP2R2C蛋白表达水平分别为2.15±0.10和0.92±0.03,与NC组比较,差异有统计学意义(P<0.01)。miR-572和PPP2R2C为靶向关系。结论 miR-572负向调控PPP2R2C抑制鼻咽癌细胞增殖。
Objective To investigate the effect of microRNA-572(miR-572)targeting regulation of protein phosphatase 2 regulatory subunit 2C(PPP2R2C)on the proliferation of nasopharyngeal carcinoma cells and its mechanism.Methods CNE cells were divided into control group,NC group(negative control),PPP2R2C group(transfection PPP2R2C),miR-in group(transfection miR-572 inhibitor)and miR-in/siPPP2R2C group(transfection miR-572 inhibitor and PPP2R2C-siRNA).Cell proliferation was detected by cell counting kit 8(CCK-8).Cell colony formation ability was detected by clonal formation assay.The percentage of cells in different cell cycles was detected by flow cytometry.The protein expression level of PPP2R2C was detected by Western blot.The targeted regulatory relationship between miR-572 and PPP2R2C was predicted and verified by Targetscan online analysis database and dual luciferase reporter genes,respectively.Results The cell proliferation activity of control group,NC group,PPP2R2C group,miR-in group and miR-in/siPPP2R2C group was(94.68±4.97)%,(93.85±3.88)%,(67.63±5.95)%,(52.37±5.60)%and(95.85±2.54)%,respectively;cell clone formation rates were(81.32±2.56)%,(80.82±4.32)%,(57.63±5.95)%,(51.54±3.48)%and(79.47±4.37)%;G0/G1 phase ratios of cells were(40.59±1.84)%,(41.06±1.94)%,(59.36±2.02),(63.47±2.24)%and(39.32±1.01)%;S phase proportion was(38.30±1.01)%,(36.25±1.08)%,(20.69±1.15)%,(16.01±1.07)%and(39.71±1.50)%.Compared with control group and NC group,cell proliferation activity,cell clonogenesis rate and the proportion of S-phase cells in PPP2R2C and miR-in groups were significantly decreased.The proportion of GO/Gl phase cells was significantly increased,with statistical significance(all P<0.01);there were no significant differences in miR-in/siPPP2R2C groups(all P>0.05).The protein expression levels of PPP2R2C in miR-in group and NC group were 2.15±0.10 and 0.92±0.03,respectively,and the dfference between two groups was statistically significant(P<0.01).miR-572 and PPP2R2C have a targeting relationship.Conclusion
作者
桂金秋
蔡克瑞
GUI Jin-qiu;CAI Ke-rui(School of Basic Medicine,Mudanjiang Medical University,Mudanjiang 15701l,Heilongjiang Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第18期2645-2649,共5页
The Chinese Journal of Clinical Pharmacology
