【摘要】 目的 探讨丝氨酸蛋白激酶ATM/CXC型趋化因子配体12(CXCL12)在去势耐受性前列腺癌细胞诱发的巨噬细胞M2极化中的作用。方法 选取人前列腺癌细胞C4-2为研究对象，使用pcDNA3.1将CXLC12过表达和/或使用小干扰RNA(siRNA)将ATM敲除后，分别与单核细胞系THP-1共同培养后，Western blot检测ATM和CXCL12水平，Transwell实验评估C4-2细胞的侵袭和迁移能力及其对巨噬细胞的募集效果，定量PCR检测M2巨噬细胞表型标志物的mRNA水平。结果 将ATM敲除的C4-2细胞与THP-1细胞共同培养后，C4-2细胞的侵袭和迁移能力受到明显抑制(P<0.05);但在将CXLC12过表达载体同时转染细胞后，ATM siRNA的抑制作用消失。进一步研究显示，伴随着ATM的敲除，抗炎标志物趋化因子配体22和白介素12亚基p40以及炎症因子转化生长因子β和白介素10基因的表达水平均降低(P<0.05),伴有巨噬细胞募集和M2极化受抑制；此作用在将CXCL12过表达后消失。结论 ATM/CXCL12信号通路的活化可通过使肿瘤微环境中M2表型躲避免疫抑制，进而促进去势耐受性前列腺癌细胞的侵袭和迁移。更多还原

【Abstract】 Objective To explore the role of serine-threonine kinase ATM/CXC chemokine ligand 12(CXCL12) in macrophage M2 polarization induced by castrated tolerant prostate cancer cells. Methods Human prostate cancer cell line C4-2 was selected as the object of study. After transfection of pcDNA3.1 to overexpress CXLC12 and/or small interfering RNAC(siRNA) to silence ATM, C4-2 cells were co-cultured with monocyte line THP-1. Western blot was used to detect levels of ATM and CXCL12. Transwell assay was applied to evaluate the invasion and migration abilities of C4-2 cells and their recruitment effect on macrophages. The mRNA levels of phenotypic markers in M2 macrophages were detected by quantitative PCR. Results After knocking out ATM in C4-2 cells and co-culturing with THP-1 cells, the invasion and migration abilities of C4-2 cells were significantly inhibited(P<0.05). However, after simultaneously transfecting CXLC12 into cells, the inhibitory effect of ATM siRNA disappeared. Further research had shown that with the knockout of ATM, levels of anti-inflammatory markers CC chemokine ligand 22 and p40 subunit of interleukin 12, as well as the inflammatory factor transforming grouth factor β and interleukin 10, were down-regulated(P<0.05), accompanied by inhibition of macrophage recruitment and M2 polarization. This effect disappeared after transfection with CXCL12. Conclusion Activation of the ATM/CXCL12 signaling pathway can promote the invasion and migration of castration tolerant prostate cancer cells by enabling the M2 phenotype in the tumor microenvironment to evade immune suppression.更多还原