Gene cloning and expression analysis of Gro/KC by primary hepatocytes

Abstract

Abstract Background and purpose: It is now well established that several environmental stresses lead to activation of p38 MAP kinase and JNK in various cell systems which is followed by chemokine producion. We investigated the expression of both CXC chemokines SDF-1a (ELR-) and Gro/KC (ELR+) in rat H4 hepatoma cells in response to heat shock, hyper-osmolarity and oxidative stresses. The pattern of expression of these chemokines by hepatoma cells in response to stress conditions was also studied.Materials and Methods: Hepatoma cells were maintained in MEM medium. Cells were subjected to different stresses [(H2O2 0.15% (w/v), manitol and NaCl (160 mM) and heat shock [(42 °C for 20 minutes)]. At the indicated time points, cells were harvested and RNA was extracted, purified and expression of the chemokines were analysed by RT-PCR. cDNA was separated by gel electrophoresis on a 1% (w/v) agarose gel and visualised on a UV transilluminator.Results: Results obtained in this report showed that there was detectable but low expression of chemokines in H4 hepatoma cells. Heat shock failed to induce expression of chemokines in H4 rat hepatoma cells. Hyper-osmolarity also has not stimulated Chemokines expression. In this study we have also shown that oxidative stress did not induce expression of chemokines. Overall, although detection is possible but regulatory responses were not observed in H4 hepatoma cells.Conclusion: Several known injurious conditions cause recruitment of macrophages, neutrophils and other immune cells to the liver. Immune cells are recruited to the hepatic vasculature following local liver injury and consequent chemokine production. Our results demonstrated that failure in production of these chemokines by Hepatoma cells may be a way to escape from immune surveillance.

Keywords

• Chemokines
• stress
• expression
• hepatoma