【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。更多还原

【Abstract】 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 200 μmol/L BER on the proliferation of OSRC-2 cell after treatment for 24 h and 48 h. After treated with 0(control group), 50, 100 μmol/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, 100 μmol/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, 100 μmol/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.更多还原