摘要
目的探究番泻苷B对视网膜母细胞瘤HXO-Rb44细胞增殖、凋亡、侵袭及Wnt/β-catenin信号通路的影响。方法通过采用不同剂量(0、5、10、20μmol/L)番泻苷B处理HXO-Rb44细胞,将细胞随机分为4组:Control组,番泻苷B 5、10、20μmol/L组。MTT法检测细胞活力;克隆形成实验检测克隆形成率;流式细胞仪检测细胞凋亡率;Transwell检测侵袭细胞数;细胞成球实验检测细胞成球直径和细胞成球数目;蛋白质印迹检测cleaved caspase-3、caspase-3、MMP-2、MMP-9、SOX2、OCT4、CD44、Wnt1、β-catenin蛋白表达。结果与Control组比较,番泻苷B 10、20μmol/L组细胞活力、克隆形成率显著降低(P<0.05),细胞凋亡率和cleaved caspase-3/caspase-3表达显著升高(P<0.05),侵袭细胞数和MMP-2、MMP-9蛋白表达显著降低(P<0.05),细胞成球直径、细胞成球数目和SOX2、OCT4、CD44蛋白表达显著降低(P<0.05),Wnt1、β-catenin蛋白表达显著降低(P<0.05)。结论番泻苷B可抑制视网膜母细胞瘤HXO-Rb44细胞增殖、侵袭和干细胞样特性,诱导细胞凋亡,抑制Wnt/β-catenin信号通路的活化。
Objective To investigate the effect of sennoside B on proliferation,apoptosis,invasion and Wnt/β-catenin signaling pathway of retinoblastoma HXO-Rb44 cells.Methods HXORB44 cells were treated with different doses(0,5,10 and 20μmol/L)of sennoside B,and the cells were randomly divided into four groups:Control group,sennoside B 5μmol/L group,sennoside B 10μmol/L group and sennoside B 20μmol/L group.Cell viability was detected by MTT assay.Colony formation assay was used to detect the colony formation rate.Cell apoptosis rate was detected by flow cytometry.Cell invasion were detected by Transwell assay.Cell pellet-forming experiment was used to detect the diameters and numbers of cell pellets.The protein expression levels of Cleaved Caspase-3,Caspase-3,MMP-2,MMP-9,SOX2,OCT4,CD44,Wnt1,β-catenin were detected by Western blotting.Results The cell viabilities and colony formation rates in the sennoside B 10 and 20μmol/L groups were significantly decreased when compared with those in the Control group(P<0.05),the cell apoptosis rates and cleaved Caspase-3/Caspase-3 expression levels were significantly increased when compared with those in the Control group(P<0.05).The numbers of invaded cells and the protein expression levels of MMP-2 and MMP-9 were significantly decreased in the sennoside B 10 and 20μmol/L groups(P<0.05),the cell pellet diameters,the cell pellet numbers and the protein expression levels of SOX2,OCT4 and CD44 were significantly decreased(P<0.05),and the protein expression levels of Wnt1 andβ-catenin were significantly decreased in the sennoside B 10 and 20μmol/L groups when compared with those in the Control group(P<0.05).Conclusion Sennoside B can induce apoptosis,inhibit the proliferation,invasion and stem-like characteristics of retinoblastoma HXO-Rb44 cells,and inhibit the activation of Wnt/β-catenin signaling pathway.
作者
孙蒙蒙
崔博坤
贾梦
冉柳
王丹荣
冯素婷
张虎
郝建章
SUN Mengmeng;CUI Bokun;JIA Meng;RAN Liu;WANG Danrong;FENG Suting;ZHANG Hu;HAO Jianzhang(Operating Room,Baoding First Hospital,Baoding,Hebei,071000,China;Department of Ophthalmology,Baoding First Hospital,Baoding,Hebei,071000,China)
出处
《医学分子生物学杂志》
CAS
2024年第2期141-145,153,共6页
Journal of Medical Molecular Biology
基金
河北省重点研发计划项目(No.223777105D)
保定市科技计划项目(No.2041ZF240)。
