[关键词]
[摘要]
【目的】 观察红景天苷抗肝癌机制。【方法】 将 H22肝癌细胞分为对照组(仅更换新鲜培养液)及红景天苷低、中、高剂 量组,相应干预48 h。细胞计数试剂盒(CCK-8)法检测细胞增殖活性,Transwell实验检测侵袭细胞数,单丹磺酰尸胺(MDC) 染色法观察细胞自噬小体数量,实时定量聚合酶链反应(qRT-PCR)法和蛋白免疫印迹(Western Blot)法相应检测细胞高迁移 率族蛋白1(HMGB1),自噬相关基因酵母Atg6同系物(Beclin1)、微管相关蛋白1A/1B-轻链3(LC3Ⅱ/Ⅰ)、选择性自噬接头蛋 白62(p62)mRNA、蛋白表达量。【结果】 与对照组比较,红景天苷低、中、高剂量组细胞增殖活性降低,侵袭细胞数和自噬 小体数减少,p62 mRNA和蛋白表达水平升高,HMGB1、LC3-Ⅱ/Ⅰ和Beclin1 mRNA和蛋白表达水平降低,差异均有统计学 意义(P < 0.05),且呈剂量依赖性。【结论】 红景天苷可能通过下调HMGB1表达及自噬水平抑制肝癌细胞增殖和侵袭。
[Key word]
[Abstract]
Objective To observe the mechanism of anti-hepatocellular carcinoma of salidroside. Methods H22 hepatocellular carcinoma cells were divided into control group(only fresh culture medium was replaced)and salidroside low-,medium- and high- dose groups,and the corresponding intervention was given in each group for 48 hours. Cellular proliferation activity was detected by cell counting kit(CCK)- 8,the number of invasive cells was detected by Transwell assay, the number of autophagosomes was observed by using dansylcadaverine (MDC)staining method,the mRNA and protein expression levels of high mobility group protein 1(HMGB1)and autophagy-related genes yeast Atg6 homolog(Beclin1), microtubule-associated protein 1A/1B-light chain 3 (LC3Ⅱ/Ⅰ)and sequestosome 62(p62)were detected accordingly by real-time quantitative polymerase chain reaction (qRT-PCR)and Western Blot,respectively. Results Compared with the control group,the cellular proliferation activity was decreased,the number of invading cells and autophagosomes were reduced in salidroside low-,mediumand high-dose groups,mRNA and protein expression level and of p62 were increased,and mRNA and protein expression levels of HMGB1,LC3-Ⅱ/Ⅰ and Beclin1 were decreased,the differences being statistically significant (P < 0.05),showing a dose-dependent manner. Conclusion Salidroside may inhibit the proliferation and invasion of hepatocellular carcinoma cells by down-regulating the expression of HMGB1 and autophagy level.
[中图分类号]
R285.5
[基金项目]
河南省高等学校重点科研项目(编号:23B320006)