BC047440通过ERK1/2-β-catenin信号通路调控肝癌干细胞分化的分子机制研究  

Molecular mechanism of BC047440 in regulating the differentiation of liver cancer stem cells though ERK1/2-β-Catenin signaling pathway

作　　者：尤楠 王峥 吴柯 王梁 YOU Nan;WANG Zheng;WU Ke;WANG Liang(Department of Hepatobiliary Surgery,Xinqiao Hospital of Army Military Medical University,Chongqing 400037,China)

机构地区：[1]陆军军医大学新桥医院肝胆外科,重庆400037

出　　处：《重庆医学》2023年第11期1627-1632,共6页Chongqing medicine

基　　金：重庆市基础科学与前沿技术研究专项项目(cstc2017jcyjAX0348)。

摘　　要：目的探讨BC047440与细胞外信号调节激酶1/2(ERK1/2)-β-连环蛋白(β-catenin)信号通路之间的关系及其对肝癌干细胞(LCSCs)分化的调控作用。方法采用shRNA慢病毒技术抑制LCSCs中BC047440的表达,实验用LCSCs分为BC047440抑制组(sLCSCs组),空白组(LCSCs组)。在指定时间点,利用酶联免疫吸附试验(ELISA)分别检测两组细胞血清清蛋白(ALB)和甲胎蛋白(AFP)表达水平,肝细胞合成代谢功能采用糖原染色实验和吲哚菁绿摄取实验检测,电镜检测两组细胞超微结构形态学变化,采用Western blot检测各组细胞中ERK1/2、磷酸化ERK1/2(p-ERK1/2)、β-catenin蛋白表达水平。通过激酶特异抑制剂PD98059抑制sLCSCs组中ERK1/2的活性后,观察β-catenin、ALB和AFP表达水平。结果shRNA慢病毒处理可以抑制LCSCs组中BC047440表达;抑制BC047440表达后,LCSCs组向成熟细胞分化导致干细胞特性减弱。进一步分子机制的研究证实,shRNA抑制BC0474407后,p-ERK1/2表达上升、β-catenin表达下降,进一步抑制sLCSCs组中ERK1/2的活性后,可上调β-catenin表达,部分恢复LCSCs组干细胞特性。结论BC047440可参与调控LCSCs分化,该分化过程可能通过ERK1/2-β-catenin信号通路实现。Objective To investigate the relationship between BC047440 and extracellular-regulated kinase 1/2(ERK1/2)-β-catenin signaling pathway as well as its regulatory effect on the differentiation of liver cancer stem cells(LCSCs).Methods The expression of BC04744 in LCSCs was inhibited using shRNA lentiviruses.The experimental LCSCs were divided into the BC047440 inhibition group(sLCSCs)and the blank group(LCSCs).At the designated time points,the changes in serum albumin(ALB)and alpha fetoprotein(AFP)expressions of both groups were detected through enzyme-linked immunosorbent assay(ELISA).Subsequently,the anabolic function of liver cells was determined with glycogen staining and indocyanine green uptake,while the ultrastructural morphological changes of cells in both groups were observed using electron microscopy and the protein expressions of ERK1/2,p-ERK1/2 andβ-catenin in each group were measured by Western blot.After the inhibition of ERK1/2 activity in sLCSCs using kinase-specific inhibitor PD98059,the changes inβ-catenin,ALB and AFP expressions were observed.Results shRNA lentivirus treatment could inhibit the expression of BC047440 in LCSCs.After the inhibition of BC047440 expression,LCSCs differentiated into mature cells,leading to the weakeni ng of stem cell characteristics.Further study on the molecular mechanism confirmed that after shRNA inhibited BC0474407,the expression of p-ERK1/2 increased while that ofβ-catenin decreased.Following the further inhibition of ERK1/2 activity in sLCSCs,the expression ofβ-catenin was up-regulated,and the characteristics of stem cells in the LCSCs group were partially restored.Conclusion BC047440 can participate in the regulation of LCSCs differentiation,which may be mediated via the ERK1/2-β-catenin signaling pathway.

关 键 词：肝癌干细胞 BC047440 细胞外信号调节激酶1/2 Β-连环蛋白 分化 

分 类 号：R735.7[医药卫生—肿瘤]