摘要
目的 探讨小檗碱(BER)对人肾癌细胞侵袭、迁移的抑制作用及可能机制。方法 以人肾癌OSRC-2细胞为对象进行实验。采用alamarBlue法测定0(对照组)、25、50、75、100、125、150、175、200μmol/L BER作用24、48 h后对OSRC-2细胞增殖的抑制作用。用0(对照组)、50、100μmol/L BER作用于细胞48 h后,通过流式细胞术分析BER对细胞周期的影响;通过细胞划痕实验观察24、36 h时各组细胞的迁移能力;通过Transwell实验观察作用24 h时各组细胞的侵袭能力;并通过Western blot法检测上述药物作用48 h后细胞中甲基转移酶样蛋白3(METTL3)的蛋白表达,通过RNA甲基化定量试剂盒检测各组细胞内RNA的N6-甲基腺嘌呤(m6A)水平。结果 与对照组比较,不同浓度BER均可显著升高OSRC-2细胞的增殖抑制率(P<0.01),并呈时间和浓度依赖性趋势;50、100μmol/L BER作用48 h后可将细胞周期阻滞在G0/G1期(P<0.01);50、100μmol/L BER组细胞的迁移、侵袭能力均显著降低(P<0.05或P<0.01),细胞中METTL3蛋白的表达水平和RNA的m6A水平均显著降低(P<0.01)。结论 BER可通过下调METTL3的表达,进而抑制m6A水平,从而抑制人肾癌细胞的侵袭、迁移。
OBJECTIVE To investigate the inhibitory effect of berberine(BER)on the invasion and migration of human renal carcinoma cells and its potential mechanism.METHODS Using human renal carcinoma OSRC-2 cell as object,alamarBlue assay was adopted to detect the inhibitory effects of 0(control group),25,50,75,100,125,150,175 and 200μmol/L BER on the proliferation of OSRC-2 cell after treatment for 24 h and 48 h.After treated with 0(control group),50,100μmol/L BER for 48 h,the effect of BER on cell cycle was analyzed by flow cytometry.The migration of OSRC-2 cells in 24 h and 36 h was observed by cell scratch test,and the invasion ability of OSRC-2 cells in 24 h was detected by Transwell assay.The protein expression of methyltransferase-like 3(METTL3)was detected by Western blot after treatment for 48 h,and RNA methylation quantification kit was used to detect the levels of N6-methyladenosine(m6A)in OSRC-2 cells.RESULTS Compared with control group,BER at different concentrations could significantly decrease the survival rate of OSRC-2 cells(P<0.01),and showed a dose-dependent and time-dependent manner.After 48 h of BER treatment at 50,100μmol/L,the cell was arrested in G0/G1 phase(P<0.01).Compared with control group,the migration and invasion abilities of cells in 50,100μmol/L BER group were significantly decreased(P<0.05 or P<0.01);the protein expression of METTL3 and the level of m6A in RNA were significantly decreased(P<0.01).CONCLUSIONS BER can inhibit level of m6A by down-regulating the expression of METTL3,thereby inhibiting the invasion and metastasis of human renal carcinoma cells.
作者
杨天宇
帅立雄
许竞泽
程馨阳
许志明
张永胜
YANG Tianyu;SHUAI Lixiong;XU Jingze;CHENG Xinyang;XU Zhiming;ZHANG Yongsheng(Dept.of Pathology,the Second Affiliated Hospital of Soochow University,Jiangsu Suzhou 215004,China)
出处
《中国药房》
CAS
北大核心
2023年第15期1863-1868,共6页
China Pharmacy
基金
苏州市科技发展项目(No.SYS2020146)
苏州市医学重点扶持学科(No.SZFCXK202137)。
