Li Jie, Yan Kun, Yang Yi, Li Hua, Wang Zhidong, Xu Xin
Department of General Surgery, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, 710004.
Nan Fang Yi Ke Da Xue Xue Bao. 2019 Dec 30;39(12):1436-1442. doi: 10.12122/j.issn.1673-4254.2019.12.07.
To investigate the regulatory role of Musashi-1 (MSI1) in the proliferation and growth of hepatocellular carcinoma (HCC) cells.
We examined the expression of MSI1 in HCC and paired adjacent tissues from 24 patients using immunohistochemistry and Western blotting. A MSI1-expressing vector was constructed and stably transfected into HepG2 cells, and short hairpin RNAs (shRNAs) that targeted MSI1 mRNA were ligated into the vector and stably transfected in Huh7 cells. The effects of MSI1 overexpression and silencing on the proliferation, viability and cell cycle of HepG2 cells were investigated using flow cytometry or MTT assay. The expressions of PCNA, cyclin D1, APC and β-catenin in the HCC cells were detected with Western blotting.
MSI1 expression was significantly up-regulated in HCC tissues as compared with that in the adjacent tissues. Overexpression of MSI1 in HepG2 cells resulted in significantly enhanced cell growth ( < 0.01) and significantly reduced G0/G1 phase cells from (58.42±3.18)% to (40.67±1.22)% and increased S phase cells from (28.51± 1.93)% to (40.06±1.92)% ( < 0.01), causing also increases in the expressions of PCNA and Cyclin D1. Knockdown of MSI1 in Huh7 cells obviously inhibited the cell growth and caused cell cycle arrest at the G1/S phase ( < 0.01) with reduced protein expressions of PCNA and cyclin D1. Overexpression of MSI1 in HepG2 cells also down-regulated the expression of APC and up-regulated the expression of β-catenin protein, while MSI1 knockdown caused reverse changes in Huh7 cells.
MSI1 promotes the progression of HCC through positive modulation of cell growth and cell cycle the Wnt/β-catenin pathway.
探讨Musashi-1（MSI1）在肝细胞癌（HCC）细胞增殖和生长中的调控作用。
我们采用免疫组织化学和蛋白质印迹法检测了24例患者肝癌组织及配对癌旁组织中MSI1的表达。构建了表达MSI1的载体并稳定转染至HepG2细胞，将靶向MSI1 mRNA的短发夹RNA（shRNAs）连接到载体中并稳定转染至Huh7细胞。使用流式细胞术或MTT法研究MSI1过表达和沉默对HepG2细胞增殖、活力和细胞周期的影响。通过蛋白质印迹法检测肝癌细胞中PCNA、细胞周期蛋白D1、APC和β-连环蛋白的表达。
与癌旁组织相比，肝癌组织中MSI1表达显著上调。HepG2细胞中MSI1过表达导致细胞生长显著增强（<0.01），G0/G1期细胞从（58.42±3.18）%显著减少至（40.67±1.22）%，S期细胞从（28.51±1.93）%增加至（40.06±1.92）%（<0.01），同时PCNA和细胞周期蛋白D1的表达也增加。Huh7细胞中MSI1敲低明显抑制细胞生长并导致细胞周期停滞在G1/S期（<0.01），PCNA和细胞周期蛋白D1的蛋白表达降低。HepG2细胞中MSI1过表达还下调了APC的表达并上调了β-连环蛋白的表达，而MSI1敲低在Huh7细胞中导致相反的变化。
MSI1通过正向调节细胞生长和细胞周期以及Wnt/β-连环蛋白途径促进肝癌进展。
