Effect Of Mice Immune Cells Stimulation On Glucose Metabolic Activities And Levels Of HepG2 Cells

Objective:In this work we investaged glucose metabolic activities and malignant developments of HepG2 cells in the microenvironment caused by mouse immune cells, and evaluated the application of detecting changes of glucose metabolic activities and co-expression of CD44 and IL-6 in hepatoma Identification.Methods:We used immune cells to treat HepG2 cells and observed their effect on glucose metabolism of HepG2 cells. Glucose detection kit was used to detect the glucose consumption rate, gas chromatography to detect the lactate production rate, qPCR to detect the mRNA level of HK2, PDK1, PFK2, HIF-1α, etc associated with the glycolytic, and immunocytochemistry to detect the protein level of HIF-3α, CD44, MMP-9, etc associated with the malignant nature of liver cancer. We performed soft agar colony formation assay to study the ability of self-renew in vitro and MTT to detect cell resistance to drugs. We used ELISA, qPCR etc to detect the expression levels of IL-6, IL-6R and GP130, and used immunohistochemistry to verify the expression levels of CD44 and IL-6 in malignant hepatoma tissues.Results:1. Mouse immune cells, included peritoneal and spleen cells can increase the rate of glucose consumption and lactate production through increasing the expression of some key glycolytic enzymes in HepG2 cells.2. Mouse immune cells can up-regulate the expression of HIF-3α, CD44, MMP-9, and TGF-β1, which is related to malignant devopment of cancers, and decrease the expression of FN. The stimulation mediated by mouse immune cells enhance the self-renewal ability and drug resistance to paclitaxel of HepG2 cells.3. Mouse immune cells can induce HepG2 cells autocrine IL-6, meanwhile up-regulate the expression of IL-6R and GP130, which further activate IL-6 signaling pathway.4. In malignant hepatoma cell or tissue, CD44 and IL-6 can co-express at high levels, which can be used to identify clinically malignant hepatoma.5. Doxorubicin may have a better therapeutic effect on inflammatory-related hepatoma.Conclusion: The microenvironment caused by mouse immune cells can promote glycolysis and malignant development of HepG2 cells. The promotaion may be involved with interaction between various immune cells or cytokines and cancer cells, while IL-6 may be the important factors of them. This study demonstrates that mouse immune cells can induce HepG2 cells to secrete IL-6, and promote glycolysis and malignant progression of HepG2 cells. The study also showed that high levels of co-expression of CD44 and IL-6 may be used to identify hepatoma, and doxorubicin exibited a better therapeutic effect on inflammatory-related hepatoma. These results have certain reference significance for the clinical assessment and treatment of malignant hepatoma.