Cheng Bianqiao, Huang Zhiteng, Wu Linlin, Lin Yanchen, Lin Weiguo, Zhu Qi
Department of Gastroenterology, The Second Hospital of Fuzhou Affiliated Xiamen University, Fuzhou, Fujian, China.
J BUON. 2021 Mar-Apr;26(2):435-443.
The purpose was to investigate the effect of activated human hepatic stellate cell (HSC) microenvironment on the metastatic capacity of hepatocellular carcinoma (HCC) cells and its underlying mechanism.
LX-2 HSCs were stimulated with Human Transforming Growth Factor-Beta 1(TGF-β1), and protein expression of α-smooth muscle actin (α-SMA) and filamentous actin (F-actin) were determined to verify the activation of LX-2 cells. Next, SMMC7721 HCC cells were cultured in the conditioned medium originating from activated LX-2 cells. Wound healing and Transwell assays were performed to examine cell migration and invasion. The expression of metastasis-related genes Matrix Metalloproteinase9 (MMP9), N-cadherin, and Vascular endothelial growth factor (VEGF) was detected. ELISA was carried out to determine the interleukin (IL) -1β level. Finally the inhibitors of TGF-β1 and IL-1β were employed to investigate the roles of LX-2 activation and IL-1β in the metastasis-related gene alterations.
TGF-β1 activated LX-2 cells, as evidenced by up-regulated α-SMA and F-actin expression. Compared with the control medium, the conditioned medium derived from LX-2 cells significantly promoted the migration and invasion of SMMC7721 cells. And it also up-regulated mRNA and protein expression of the metastasis-related genes in SMMC7721 cells. Furthermore, it resulted in a significant increase in the IL-1β level in SMMC7721 cells. Importantly, TGF-β1 inhibitor and IL-1β inhibitor either individually or synergistically abolished the up-regulated expression of conditioned medium-induced metastasis-related gene in SMMC7721 cells.
The conditioned medium generating from TGF-β1-activated LX2 cells can enhance the metastatic ability of SMMC7721 cells through up-regulating IL-1 expression.
研究活化的人肝星状细胞（HSC）微环境对肝癌（HCC）细胞转移能力的影响及其潜在机制。
用人类转化生长因子-β1（TGF-β1）刺激LX-2肝星状细胞，检测α-平滑肌肌动蛋白（α-SMA）和丝状肌动蛋白（F-actin）的蛋白表达，以验证LX-2细胞的活化。接下来，将SMMC7721肝癌细胞培养于源自活化LX-2细胞的条件培养基中。进行伤口愈合实验和Transwell实验以检测细胞迁移和侵袭能力。检测转移相关基因基质金属蛋白酶9（MMP9）、N-钙黏蛋白和血管内皮生长因子（VEGF）的表达。采用酶联免疫吸附测定（ELISA）法测定白细胞介素（IL）-1β水平。最后，使用TGF-β1和IL-1β抑制剂研究LX-2活化和IL-1β在转移相关基因改变中的作用。
TGF-β1激活了LX-2细胞，表现为α-SMA和F-actin表达上调。与对照培养基相比，源自LX-2细胞的条件培养基显著促进了SMMC7721细胞的迁移和侵袭。并且它还上调了SMMC7721细胞中转移相关基因的mRNA和蛋白表达。此外，它导致SMMC7721细胞中IL-1β水平显著升高。重要的是，TGF-β1抑制剂和IL-1β抑制剂单独或协同作用均消除了条件培养基诱导的SMMC7721细胞中转移相关基因的上调表达。
由TGF-β1激活的LX2细胞产生的条件培养基可通过上调IL-1表达增强SMMC7721细胞的转移能力。
