黄柏酮对肺癌细胞核糖体合成、增殖的影响及机制

Effects of obacunone on ribosome biogenesis and cell proliferation of lung cancer cells and the mechanism

目的探讨黄柏酮对非小细胞肺癌细胞(A549细胞)增殖的影响及潜在的作用机制。方法常规培养A549细胞,将对数生长期A549细胞随机分为对照组、黄柏酮组,分别加入DMSO溶剂、50μmol/L黄柏酮继续培养。用尿嘧啶核苷类似物(EU)标记法检测细胞核糖体RNA(rRNA)合成,用胸腺嘧啶核苷类似物(EdU)标记法检测细胞增殖能力,用实时荧光定量PCR检测细胞中rRNA(5.8S rRNA、18S rRNA、28S rRNA、45S pre-rRNA)表达,用West‑ern blotting法检测细胞中磷酸化的雷帕霉素靶蛋白(p-mTOR)、磷酸化的S6核糖体蛋白235/236[p-RPS6(235/236)]、p-RPS6(240/244)蛋白表达。结果黄柏酮组荧光信号强度与对照组荧光信号强度的比值为0.684±0.008(P<0.05)。与对照组比较,黄柏桐组EdU阳性细胞率低,5.8S rRNA、18S rRNA、28S rRNA、前体45S rRNA相对表达量低,p-mTOR、p-RPS6(235/236)、p-RPS6(240/244)蛋白相对表达量低(P均<0.05)。结论黄柏酮可抑制肺癌细胞核糖体合成及增殖,其作用机制可能与抑制mTOR/RPS6信号通路激活有关。

Objective To investigate the effect of obacunone on the proliferation of non-small-cell lung cancer cells A549 and its potential mechanism.Methods A549 cells in the logarithmic growth phase were randomly divided into the control group and the obacunone treatment group,which were added with DMSO solvent and 50μmol/L obacunone,re‑spectively,and we continued to culture them.We used EU labeling method to detect cell ribosomal RNA(rRNA)synthe‑sis,and EdU labeling method to detect cell proliferation ability.Real-time fluorescent quantitative PCR was used to detect the expression of rRNA(5.8S rRNA,18S rRNA,28S rRNA,45S pre-rRNA)in cells,and Western blotting was used to detect the protein expression of p-mTOR,p-RPS6(235/236),and p-RPS6(240/244)in cells.Results The ratio of the fluorescence signal intensity of the obacunone group to that of the control group was 0.684±0.008(P<0.05).Com‑pared with the control group,the rate of EdU positive cells in the obacunone treatment group was lower(P<0.05).Com‑pared with the control group,the relative expression levels of 5.8S rRNA,18S rRNA,28S rRNA,and pre-45S rRNA de‑creased,and the relative expression levels of p-mTOR,p-RPS6(235/236),and p-RPS6(240/244)proteins decreased in the obacunone group(all P<0.05).Conclusion Obacunone can inhibit ribosome synthesis and proliferation of lung cancer cells,and its mechanism may be related to the inhibition of mTOR/RPS6 signaling pathway.