肺癌细胞促进单核吞噬细胞MAPK相关通路活化诱导炎性细胞因子表达上调的研究

Study on upregulation of expression of inflammatory cytokines induced by MAPK signaling related pathway of mononuclear phagocytes promoted by lung cancer cells

目的通过建立人单核吞噬细胞系(THP-1)和人肺腺癌细胞系(SPC-A1)共培养体系,检测共培养体系中THP-1的炎性细胞因子mRNA表达水平和丝裂原活化蛋白激酶通路(MAPK)的表达情况,初步阐明肺癌微环境对THP-1中炎性细胞因子表达的影响。方法体外培养SPC-A1细胞系、THP-1细胞系,并建立二者共培养体系,设置THP-1单独培养组为对照组。24h后,收集各组THP-1细胞,实时荧光定量聚合酶链式反应(RealTime PCR)检测白细胞介素(IL)-1β、-6、-8及肿瘤坏死因子-α(TNF-α)的mRNA表达水平;Western blot检测MAPK通路蛋白c-Jun氨基末端激酶及其磷酸化形式(JNK/p-JNK)、丝裂原活化蛋白激酶p38亚单位及其磷酸化形式(p38 MAPK/p-p38 MAPK)、细胞外调节蛋白激酶及其磷酸化形式(ERK/p-ERK)的表达。结果共培养24h后,共培养组THP-1的IL-1β、IL-8mRNA表达水平均明显高于对照组(P<0.05),分别为对照组的5.81、4.21倍;Western blot结果显示共培养组p38 MAPK、p-p38MAPK的表达水平均明显高于对照组(P<0.05)。结论肺癌细胞可引起肺癌环境中单核吞噬细胞中p38 MAPK通路的活化,进而诱导IL-1β、IL-8mRNA表达上调,有利于肿瘤炎症微环境的维持和促进肿瘤的进展。

Objective To establish the co‐culture system of mononuclear phagocyte system （T HP‐1） and hu‐man lung adenocarcinoma cell line （SPC‐A1）, detect the expression of mRNA of inflammatory cytokines in the T HP 1 cell line and MAPK pathways in the co‐culture system, and preliminary indicate the effect of lung cancer microenvi‐ronment on the expression of inflammatory cytokines in THP‐1 cell line. Methods The SPC‐A1 and THP‐1 cell lines were cultured in vitro, and the co‐culture system was established. The single cultured THP‐1 cell line was setted as control group. 24 h after culturing, THP‐1 cells were collected and real‐time PCR was used to detect interleukin （IL） 1β,IL6 and IL8, and the expression of tumor necrosis factorα （TNFα） mRNA. The western blot analysis was used to detect the expression of JNK/p, JNK, p38 MAPK/p, p38MAPK, ERK/p and ERK. Results 24 h after co‐cultu‐ring, the expression levels of IL1β and IL8 in the co‐cultured group was obviously higher than those in the control group, and was 5. 81 and 4. 21 fold higher than the control group, respectively （P〈 0. 05）. Western blot analysis showed that the expression levels of p38 MAPK/p and p38 MAPK in the co‐cultured group was significant higher than those in the control group （P〈0. 05）. Conclusion The lung cancer cells could active the p38 MAPK signaling pathway in monocyte macrophage cells and induce further upregulation of the expression of IL1β,IL8 mRNA, which might be benefit for the maintenance of tumor microenvironment and tumor progression.