沉默PrxⅡ基因促进巨噬细胞对肝癌细胞的杀伤作用  

Lethal Effect of Macrophages Promoted by Silencing Prx 域 Gene on Hepatoma Cells

作　　者：冯琳 孙率洋 姜鹏 吕树旗 韩英浩[1] Feng Lin;Sun Shuaiyang;Jang Peng;Lv Shuqi;Han Yinghao(College of Life Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319)

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,大庆163319

出　　处：《黑龙江八一农垦大学学报》2024年第2期77-83,共7页journal of heilongjiang bayi agricultural university

基　　金：黑龙江省自然科学基金项目(LH2021C061)。

摘　　要：为探究PrxⅡ对RAW264.7(小鼠单核巨噬细胞系)细胞杀伤HepG2(人肝癌细胞系)细胞的影响,阐明PrxⅡ基因调控RAW264.7细胞分泌细胞因子杀伤HepG2细胞的作用,利用慢病毒干扰技术制备PrxⅡ基因沉默的RAW264.7细胞系、MTT assay测定HepG2细胞存活情况、ELISA法测定RAW264.7细胞分泌炎性因子情况、蛋白质免疫印迹法检测NF-κB信号通路相关蛋白质表达水平。结果表明:已成功制备PrxⅡ基因沉默的RAW264.7细胞系;MTT检测发现RAW264.7细胞上清液处理HepG2细胞后,shPrxⅡ组HepG2细胞存活率明显低于mock组,且呈时间依赖性;ELISA结果表明shPrxⅡ组RAW264.7细胞分泌的TNF-α含量明显高于mock组,IL-6和IL-1β含量明显低于mock组;蛋白质免疫印迹结果与mock组相比,PrxⅡ基因沉默的RAW264.7细胞中NF-κB信号通路明显被激活。由此得出结论,PrxⅡ基因能调控RAW264.7细胞分泌细胞因子,杀伤HepG2细胞,导致HepG2细胞死亡水平上升,研究结果可为以巨噬细胞为靶点的免疫疗法提供理论基础,为肝癌患者的免疫治疗提供新思路。To explore the effect of PrxⅡon RAW264.7(mouse monocyte macrophage line)cells killing HepG2(human hepatoma cell line)cells,and to clarify the role of PrxⅡgene in regulating RAW264.7 cells to secrete cytokines to kill HepG2 cells,RAW264.7 cell line with PrxⅡgene 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 PrxⅡgene silencing was successfully constructed;MTT assay showed that the survival rate of shPrxⅡHepG2 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-αsecreted by shPrxⅡRAW264.7 cells was higher than that in shPrxⅡgroup,while the content of IL-6 and IL-1βwas lower;Western blot analysis showed that NF-κB signal pathway in shPrxⅡRAW264.7 cells was obviously activated.It was concluded that PrxⅡgene 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.

关 键 词：RAW264.7细胞系 HEPG2细胞系 PrxⅡ NF-ΚB 炎性因子 

分 类 号：Q28[生物学—细胞生物学]