结核分枝杆菌通过诱导巨噬细胞M2极化促进A549细胞增殖、迁移和侵袭  

Mycobacterium tuberculosis promotes proliferation,migration and invasion of A549 lung cancer cells by inducing macrophage M2 polarization

作　　者：陈洋 杨小琦 旷华兵 叶萌 李晓倩 邓成敏 张隆泽 谭心 吴杰 Chen Yang;Yang Xiaoqi;Kuang Huabing;Ye Meng;Li Xiaoqian;Deng Chengmin;Zhang Longze;Tan Xin;Wu Jie(Scientific Research Center,the Third Affiliated Hospital of Zunyi Medical University(the First People’s Hospital of Zunyi),Zunyi Guizhou 563000,China;College of Basic Medicine,Zunyi Medical University,Zunyi Guizhou 563006,China)

机构地区：[1]遵义医科大学第三附属医院,遵义市第一人民医院中心实验室,贵州遵义563000 [2]遵义医科大学基础医学院,贵州遵义563006

出　　处：《遵义医科大学学报》2024年第11期1061-1068,共8页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:82060506);贵州省科技厅科技计划项目[NO:黔科合支撑(2020)4Y121];遵义市科技计划项目[NO:遵市科人才(2023)5]。

摘　　要：目的探讨结核分枝杆菌感染对巨噬细胞极化的影响并分析极化后的巨噬细胞对A549肺癌细胞恶性生物学行为影响。方法采用佛波酯(PMA)诱导人单核细胞THP-124 h为M0巨噬细胞,继续用IL-4、IL-13刺激M0巨噬细胞48 h向M2极化(M2),用M.tuberculosis H37Rv裂解总蛋白刺激M0巨噬细胞48 h(M0H)。WB检测M0、M2、M0H巨噬细胞IL-10、CD163、TNF-α蛋白表达水平;CCK-8、伤口愈合实验、Transwell实验检测M0、M2、M0H巨噬细胞条件培养基(CM)对A549细胞增殖、迁移和侵袭能力的影响。结果与对照M0组相比,M2极化标志物IL-10和CD163蛋白表达水平在M2组(IL-10:M0 vs M2=0.84±0.29 vs 1.76±0.34;CD163:M0 vs M2=0.84±0.16 vs 1.66±0.21)和M0H组(IL-10:M0 vs M0H=0.84±0.29 vs 1.47±0.29;CD163:M0 vs M0H=0.84±0.16 vs 1.65±0.36)中显著升高(P<0.01);与对照M0组相比,M1极化标志物TNF-α蛋白表达水平在M2组无明显变化(M0 vs M2=0.90±0.13 vs 1.04±0.07,P=0.12),在M0H组显著下降(M0 vs M0H=0.90±0.13 vs 0.65±0.10,P<0.05);与对照M0-CM组相比,M2-CM组和M0H-CM组能够增加A549细胞增殖[48 h:M0-CM vs M2-CM=(100.00±4.75)%vs(129.51±4.59)%,P<0.01;M0-CM vs M0H-CM=(100.00±4.75)%vs(152.04±8.11)%,P<0.01]、横向迁移[24 h:M0-CM vs M2-CM=(17.08±3.16)%vs(30.98±3.28)%,M0 vs M0H=(17.08±3.16)%vs(27.51±0.65)%;48 h:M0-CM vs M2-CM=(52.62±5.45)%vs(70.75±2.33)%;M0-CM vs M0H-CM=(52.62±5.45)%vs(70.91±4.29)%,P<0.01]、纵向迁移(24 h:M0-CM vs M2-CM=100.00±4.93 vs 160.00±18.73,M0 vs M0H=10.00±4.93 vs 195.00±6.24;48 h:M0-CM vs M2-CM=43.11±4.66 vs 103.56±10.62,M0 vs M0H=43.11±4.66 vs 136.56±23.49,P<0.01)和侵袭(24 h:M0-CM vs M2-CM=62.4±4.56 vs 127.2±12.39,M0 vs M0H=62.4±4.56 vs 131.40±10.88;48 h:M0-CM vs M2-CM=86.40±8.96 vs 162.40±15.95,M0 vs M0H=86.40±8.96 vs 177.00±13.11,P<0.01)。结论结核分枝杆菌感染通过诱导巨噬细胞M2极化促进A549细胞增殖、迁移和侵袭。Objective To investigate the effect of Mycobacterium tuberculosis infection on the polarization of macrophages and analyze the effect of polarized macrophages on the malignant behavior of A549 lung cancer cells.Methods Human monocytic THP-1 cells were induced into M0 macrophages by Phorbol 12-myristate 13-acetate(PMA)for 24 h,and M0 macrophage was incubated with IL-4 and IL-13 for 48 h to induce M2 macrophage polarization(M2)or stimulated with M.tuberculosis H37Rv whole-cell lysate(M0H)for 48 h.The expression levels of IL-10,CD163 and TNF-αin M0,M2 and M0H macrophages were detected by WB.The effects of M0,M2 and M0H macrophage conditioned medium(CM)on proliferation,migration and invasion of A549 cells were detected by CCK-8,wound healing assay and transwell assay.Results Compared with the control M0 group,the M2 polarisation markers IL-10 and CD163 protein expression levels were significantly increased in the M2 group(IL-10:M0 vs M2=0.84±0.29 vs 1.76±0.34;CD163:M0 vs M2=0.84±0.16 vs 1.66±0.21)and the M0H group(IL-10:M0 vs M0H=0.84±0.29 vs 1.47±0.29;CD163:M0 vs M0H=0.84±0.16 vs 1.65±0.36)(P<0.01);compared to the control M0 group,the M1 polarisation marker TNF-αprotein expression level was significantly higher did not change in the M2 group(M0 vs M2=0.90±0.13 vs 1.04±0.07,P=0.12)did not change significantly and decreased significantly in the M0H group(M0 vs M0H=0.90±0.13 vs 0.65±0.10,P<0.05);compared with the control M0-CM group,the M2-CM group and the M0H-CM group were able to increase the proliferation of A549 cells[48 h:M0-CM vs M2-CM=(100.00±4.75)%vs(129.51±4.59)%,P<0.01;M0-CM vs M0H-CM=(100.00±4.75)%vs(152.04±8.11)%,P<0.01],lateral migration[24 h:M0-CM vs M2-CM=(17.08±3.16)%vs(30.98±3.28)%,M0 vs M0H=(17.08±3.16)%vs(27.51±0.65)%;48 h:M0-CM vs M2-CM=(52.62±5.45)%vs(70.75±2.33)%;M0-CM vs M0H-CM=(52.62±5.45)%vs(70.91±4.29)%,P<0.01],vertical migration(24 h:M0-CM vs M2-CM=100.00±4.93 vs 160.00±18.73,M0 vs M0H=10.00±4.93 vs 195.00±6.24;48 h:M0-CM vs M2-CM=43.11±4.66 vs 103.56±10.62,M0

关 键 词：结核分枝杆菌 巨噬细胞极化 肺癌 细胞增殖 迁移 侵袭 

分 类 号：R378.91[医药卫生—病原生物学]