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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 a co-culture system of human mononuclear phagocyte (THP-1) and human lung adenocarcinoma cell line (SPC-A1) to detect the expression of THP-1 mRNA and cytokines in co-culture system Activation of protein kinase pathway (MAPK) expression, initially clarify the impact of lung cancer microenvironment on THP-1 expression of inflammatory cytokines. Methods SPC-A1 cell line and THP-1 cell line were cultured in vitro and co-culture system was established. THP-1 alone culture group was set as the control group. Twenty-four hours later, THP-1 cells of each group were collected and the expression of interleukin (IL) -1β, -6, -8 and tumor necrosis factor-α (TNF-α) were detected by real-time quantitative polymerase chain reaction The expression of MAPK pathway c-Jun N-terminal kinase and its phosphorylated form (JNK / p-JNK), mitogen-activated protein kinase p38 subunit and its phosphorylated form (p38 MAPK / p-p38 MAPK), extracellular regulated protein kinase and its phosphorylated forms (ERK / p-ERK). Results After co-cultured for 24 h, the expression levels of IL-1β and IL-8 mRNA in THP-1 cells in co-culture group were significantly higher than those in control group (5.81 and 4.21 times respectively) The expression levels of p38 MAPK and p-p38MAPK were significantly higher than those of the control group (P <0.05). Conclusions Lung cancer cells can induce the activation of p38 MAPK pathway in mononuclear phagocytes in lung cancer and induce the up-regulation of IL-1β and IL-8 mRNA expression, which is beneficial to the maintenance of tumor inflammation microenvironment and the promotion of tumor progression.