Lethal Effect of Macrophages Promoted by Silencing Prx Ⅱ Gene on Hepatoma Cells
To explore the effect of Prx Ⅱ on RAW264.7(mouse monocyte macrophage line)cells killing HepG2(human hepatoma cell line)cells,and to clarify the role of Prx Ⅱ gene in regulating RAW264.7 cells to secrete cytokines to kill HepG2 cells,RAW264.7 cell line with Prx Ⅱ gene silencing was prepared using lentivirus mediated siRNA interference,survival rate of HepG2 cell was detected by MTT assay,RAW264.7 cell secretion of inflammatory factors were detected by ELISA method,and related protein expression levels of NF-κB signal pathway were detected by Western blot.The results showed that RAW264.7 cell line with Prx Ⅱgene silencing was successfully constructed;MTT assay showed that the survival rate of shPrx ⅡHepG2 cells was significantly lower than that in mock group in a time dependent manner after treated with RAW264.7 cells supernatant;ELISA results showed that TNF-α secreted by shPrx Ⅱ RAW264.7 cells was higher than that in shPrx Ⅱ group,while the content of IL-6 and IL-1β was lower;Western blot analysis showed that NF-κB signal pathway in shPrx Ⅱ RAW264.7 cells was obviously activated.It was concluded that Prx Ⅱ gene could regulate RAW264.7 cells to secrete cytokines to kill HepG2 cells,leading to an increase in the death level of HepG2 cells,which could provide a basic theory for immunotherapy targeting macrophages and provide a new idea for immunotherapy of liver cancer patients.
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