The Study Of CXCL1 Induced Endoplasmic Reticulum Stress

Objective:At present, many studies suggest tumor biological characteristics and the tumor microenvironment have a close relationship. Many of the cells factor in the tumor microenvironment can promote the development of the tumor, but also because of high interstitial pressure, insufficient blood supply, the relative lack of nutrition that easy cause tumor cells endoplasmic reticulum stress and activate the related pathways, promote the cytokine expression and the development of tumor. Recently many reporting have think thdit the CXCL1 have the correlation of tumorigenesis, development, migration, invasion ect. for example, it’s over expression in melanoma, hepatic carcinoma, gastric cancer, ovarian cancer etc tumor and it promote the malignant degree of tumor. However, the mechanisms that CXCL1 promote the development, migration, invasion of tumor remain unfolding. Tumor microenvironment plays an important role in the development, migration and invasion of tumor. Therefore, in this study, study the effect of CXCL1 on tumor from the viewpoint of tumor microenvironment. The study provided a new direction for further research on the basis of CXCL1 and hepatic carcinoma therapy.Method:Firstly, HepG2 cells were stimulated with different concentrations of Tunicamycin (TM) to Induced endoplasmic reticulum stress, Using Q-PCR to detect the mRNA level of ELR+family chemokines and using ELISA to detect the level of CXCL1 in the medium. The HepG2 cells were stimulated by the tunicamycin and after 12h replaced fresh medium for it. The medium was collected after 24 hours and to induct HepG2 cells and maturation of THP-1 cells. Using Q-PCR to detect the mRNA levels of the endoplasmic reticulum stress related genes ATF-6, PERK and IRE1. Synthesis SiCXCL1 sequence and transiently transfected into HepG2 cells to knock down the expression of its CXCL1, then stimulated by the tunicamycin and after 12h replaced fresh medium for it. The medium was collected after 24 hours and to induct HepG2 cells and maturation of THP-1 cells. Using Q-PCR to detect the mRNA levels of the endoplasmic reticulum stress related genes ATF-6, PERK and IRE1. Using gene recombination technique to Construct CXCL1 vector and transiently transfected into HepG2 cells to over expression. Using Q-PCR to detect the mRNA levels of the endoplasmic reticulum stress related genes ATF-6, PERK and IRE1. All of the above data are analyzed by SPSS 16.0 software, the results were compared by one-way ANOVA, P<0.05 has the statistical significance.Results:Under different concentrations of tunicamycin, the mRNA levels of CXCL1,CXCL2 and CXCL3 were higher than those in control group (P<0.05), CXCL8 reduction, CXCL5, CXCL6, CXCL7, CXCLR1, CXCLR2 was not detected in HepG2 cells. The levels of CXCLlis elevated in the culture medium. When HepG2 cells and maturation of THP-1 cells were stimulated by Culture medium, the mRNA of the endoplasmic reticulum stress related genes ATF-6, PERK and IRE1’s levels were increased (P<0.05).When CXCL1 is knocked down, the mRNA of the endoplasmic reticulum stress related genes ATF-6, PERK and IRE1’levels were reduced (P<0.05). CXCL1 vector was constructed successfully and expression, When CXCL1 is over expressed, the mRNA of the endoplasmic reticulum stress related genes ATF-6, PERK and IREl’s levels were increased.Conclusion:In hepatic carcinoma microenvironment, the cells are in the endoplasmic reticulum stress can promote the expression of CXCL1, CXCL2, CXCL3. While ER-stress can was caused by CXCL1 to through autocrine or paracrine. They promote each other.