The Effect of Chemotactic Factor CXCL1 Secreted by Hepatic Stellate Cells on Hepatic Carcinoma Outcomes

Abstract: This research investigated the influence of human hepatic stellate cells (HSC) on the migration， invasion and EMT of hepatocellular carcinoma (HCC) cells and underlying mechanism. HCC cells (HepG2， SMMC-7721) were co-cultured with conditional medium of HSC or HSC themselves. Cell migration and invasion were detected by methods of cell wound healing and Transwell chamber assays. Product of CXCL1 in HSC and HSC-conditional medium (HSC-CM)， the expression of CXCR2 (CXCL1 receptor 2) in HCC cells， and p-PI3K， p-AKT， p-GSK-3β and Snail in conditional cultivated HCC cells were detected by Western blot. The changes of epithelial markers， E-cadherin， mesenchymal markers， N-cadherin and Vimentin in HCC cells were detected by laser scanning confocal microscopy (LSCM) and Western blot. The results showed that HSC produced much chemokines CXCL1， while HCC cells (HepG2， SMMC-7721) highly expressed CXCL1 receptor CXCR2. Moreover， their morphology changed， adhesion ability decreased， migration and invasion ability enhanced， the expression of epithelial marker E-cadherin was downregulated and mesenchymal markers N-cadherin and Vimentin were up-regulated in conditional cultivated HCC cells. Furthermore， the phosphorylation levels of the important members of PI3K/AKT signal pathway， PI3K and AKT were up-regulated， as well as levels of p-GSK-3β and transcription factor Snail under conditional cultivation. In contrast， the expression of epithelial marker E-cadherin was up-regulated， but mesenchymal markers N-cadherin and Vimentin were down-regulated， and intracellular p-PI3K， p-AKT， p-GSK-3β and Snail were down-regulated， when the conditional cultivated HCC cells were treated by CXCR2 inhibitor (SB265610). Our results suggested that hepatic stellate cells promoted epithelial-mesenchymal transition of HCC cells through activating PI3K/AKT signaling pathway by CXCL1/CXCR2 axis.

CSTR: 32200.14.cjcb.2015.06.0007