CAJ | 학술논문

目的探讨肝再生磷酸酶-3(PRLd)在结肠癌微环境中通过肿瘤相关性巨噬细胞(TAMs)促进结肠癌细胞转移的机制.方法将TAMs与稳转染PRL-3的结肠癌Lovo细胞(Lovo-P)进行体外共培养的方式,再通过实时定量聚合酶链反应(Real-time PCR)筛选出TAMs中上调的细胞因子.Western blot检测细胞因子蛋白水平.免疫荧光法检测结肠癌患者肿瘤切片中相关TAMs表达,Kaplan-Meier法绘制患者的生存曲线.结果 Real-time PCR检测结果显示,TAMs与Lovo-P细胞共培养比与Lovo-C细胞共培养后白细胞介素(IL)-6 mRNA表达明显升高(8.3±2.2)倍.Western blot检测结果显示TAMs与Lovo-P细胞共培养后IL-6表达上调180％.在共培养体系中预处理IL-6中和抗体(5mg/L及10 mg/L)后,Lovo-P细胞侵袭性下降[(220±13)个比(132±12)个比(83±7)个].免疫荧光检测结果显示结肠癌患者癌灶处肿瘤切片中TAMs与IL-6荧光重合,分布于肿瘤间质中,并且Ⅰ期患者IL-6阳性TAMs数量要少于Ⅳ期患者[(24±4)个比(56±9)个].生存曲线分析发现IL-6阳性且TAMs＞ 20个的患者生存期明显短于IL-6阳性且TAMs≤20个患者(27.3个月比12.2个月).结论 PRL-3能够通过诱导TAMs分泌IL-6,促进结肠癌细胞转移,并影响患者的预后和生存.
목적 탐토간재생린산매-3(PRLd)재결장암미배경중통과종류상관성거서세포(TAMs)촉진결장암세포전이적궤제.방법 장TAMs여은전염PRL-3적결장암Lovo세포(Lovo-P)진행체외공배양적방식,재통과실시정량취합매련반응(Real-time PCR)사선출TAMs중상조적세포인자.Western blot검측세포인자단백수평.면역형광법검측결장암환자종류절편중상관TAMs표체,Kaplan-Meier법회제환자적생존곡선.결과 Real-time PCR검측결과현시,TAMs여Lovo-P세포공배양비여Lovo-C세포공배양후백세포개소(IL)-6 mRNA표체명현승고(8.3±2.2)배.Western blot검측결과현시TAMs여Lovo-P세포공배양후IL-6표체상조180％.재공배양체계중예처리IL-6중화항체(5mg/L급10 mg/L)후,Lovo-P세포침습성하강[(220±13)개비(132±12)개비(83±7)개].면역형광검측결과현시결장암환자암조처종류절편중TAMs여IL-6형광중합,분포우종류간질중,병차Ⅰ기환자IL-6양성TAMs수량요소우Ⅳ기환자[(24±4)개비(56±9)개].생존곡선분석발현IL-6양성차TAMs＞ 20개적환자생존기명현단우IL-6양성차TAMs≤20개환자(27.3개월비12.2개월).결론 PRL-3능구통과유도TAMs분비IL-6,촉진결장암세포전이,병영향환자적예후화생존.
Objective To investigate the mechanisms of phosphatase of regenerating Liver-3 (PRL-3) which promotes colon cancer cells (CRCs) metastasis through tumor associated macrophages (TAMs).Methods TAMs were cocuhured with Lovo-P cells.Real-time quantitative polymerase chain reaction (Real-time PCR) was used to screen the elevated mRNA of cytokines of the TAMs,and western blotting was used to examine the protein levels of the elevated cytokines.Immunofluorescence was used to examine the related TAMs in the sections of colon cancer patients.Kaplan-Meier was used to analysis the survival curves of the patients.Results Real-time PCR showed that the interleukin (IL)-6 mRNA of TAMs was significantly elevated (8.3 ± 2.2 fold higher),western blotting showed that the level of IL-6 elevated about 180％.After IL-6 antibody (5 mg/L and 10 mg/L) was added to the coculture medium,the invasion of Lovo-P cells was decreased significantly (220 ± 13 vs.132 ± 12 vs.83 ±7).Immunofluorescence showed that the location of TAMs was coincided with IL-6 fluorescence,and the IL-6 positive TAMs amounts of stage I patients are less than the stage Ⅳ patients (24 ±4 vs.56 ±9).A Kaplan-Meier survival curve demonstrated that patients with a high IL-6 positive TAMs counts (＞20) survive significantly shorter than those with low IL-6 positive TAMs counts (＜20) (27.3 m vs.12.2 m).Conclusion PRL-3 could induce TAMs secreting IL-6 and promotes CRCs metastasis,and influence the prognosis and survive of the patients.