CAJ | 학술논문

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（Real‐TimePCR）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；Westernblot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（p38MAPK／p‐p38MAPK）、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；Westernblot结果显示共培养组p38MAPK、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p38MAPK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。
목적：통과건립인단핵탄서세포계（THP‐1）화인폐선암세포계（SPC‐A1）공배양체계，검측공배양체계중THP‐1적염성세포인자mRNA표체수평화사렬원활화단백격매통로（MAPK）적표체정황，초보천명폐암미배경대THP‐1중염성세포인자표체적영향。방법체외배양SPC‐A1세포계、THP‐1세포계，병건립이자공배양체계，설치THP‐1단독배양조위대조조。24h후，수집각조THP‐1세포，실시형광정량취합매련식반응（Real‐TimePCR）검측백세포개소（IL）‐1β、‐6、‐8급종류배사인자‐α（TNF‐α）적mRNA표체수평；Westernblot검측MAPK통로단백c‐Jun안기말단격매급기린산화형식（JNK／p‐JNK）、사렬원활화단백격매p38아단위급기린산화형식（p38MAPK／p‐p38MAPK）、세포외조절단백격매급기린산화형식（ERK／p‐ERK）적표체。결과공배양24h후，공배양조THP‐1적IL‐1β、IL‐8mRNA표체수평균명현고우대조조（P＜0．05），분별위대조조적5．81、4．21배；Westernblot결과현시공배양조p38MAPK、p‐p38MAPK적표체수평균명현고우대조조（P＜0．05）。결론폐암세포가인기폐암배경중단핵탄서세포중p38MAPK통로적활화，진이유도IL‐1β、IL‐8mRNA표체상조，유리우종류염증미배경적유지화촉진종류적진전。
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.