Xie Yuxiao, Liao Rui, Pan Long, Fan Kai, Peng Cong, Du Chengyou
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Feb;33(2):210-4.
To investigate the influence of hepatic stellate cells( HSCs) on malignant biological behavior of hepatoma cells and related mechanisms.
Human hepatoma cell lines SMMC-7721 and Hep G2,and hepatic stellate cell line LX-2 were cultured separately. HSC conditioned medium( LX2-CM),MAPK specific inhibitor U0126 were used to treat hepatoma cells,separately or together. The invasion and migration abilities of hepatoma cells were detected by TranswellTMassay,and cell proliferation was analyzed by CCK-8 assay. The mRNA and protein expression levels of p-ERK1 /2,ERK1 /2,c-Myc,vimentin and E-cadherin were determined by real-time PCR and Western blot analysis,respectively.
LX2-CM promoted the proliferation,invasion and migration of SMMC-7721 and Hep G2 cells,and these effects were inhibited by U0126. LX2-CM up-regulated the expression levels of p-ERK1 /2,c-Myc,vimentin and down-regulated the expression level of E-cadherin. Conversely,after U0126 treatment,the expression levels of p-ERK1 /2,c-Myc and vimentin decreased significantly,while E-cadherin expression level increased.
LX2-CM could activate c-Myc via ERK1 /2signaling pathway in hepatoma cells,and consequently promote cell proliferation,invasion and migration,and also induce epithelial-mesenchymal transition.
探讨肝星状细胞（HSCs）对肝癌细胞恶性生物学行为的影响及其相关机制。
分别培养人肝癌细胞系SMMC-7721和Hep G2以及肝星状细胞系LX-2。用肝星状细胞条件培养基（LX2-CM）、丝裂原活化蛋白激酶（MAPK）特异性抑制剂U0126单独或联合处理肝癌细胞。采用TranswellTM实验检测肝癌细胞的侵袭和迁移能力，用CCK-8实验分析细胞增殖情况。分别通过实时荧光定量PCR和蛋白质免疫印迹法检测p-ERK1 /2、ERK1 /2、c-Myc、波形蛋白和E-钙黏蛋白的mRNA及蛋白表达水平。
LX2-CM促进SMMC-7721和Hep G2细胞的增殖、侵袭和迁移，而U0126可抑制这些作用。LX2-CM上调p-ERK1 /2、c-Myc、波形蛋白的表达水平，下调E-钙黏蛋白的表达水平。相反，U0126处理后，p-ERK1 /2、c-Myc和波形蛋白的表达水平显著降低，而E-钙黏蛋白表达水平升高。
LX2-CM可通过ERK1 /2信号通路激活肝癌细胞中的c-Myc，进而促进细胞增殖、侵袭和迁移，并诱导上皮-间质转化。
