肝再生磷酸酶-3诱导肿瘤相关性巨噬细胞分泌白细胞介素-6促进结肠癌细胞转移  

Phosphatase of regenerating liver-3 promotes colon cancer cells metastasis through inducing the secretion of interleukin-6 by tumor associated macrophages

作　　者：黄汉源[1] 许鹤洋[2] 来伟[2] 曾献清 张旸[2] 陈志坚[1] 李定文[1] 林峰[1] 褚忠华[2] 

机构地区：[1]广东省五华县人民医院普外科,514400 [2]中山大学孙逸仙纪念医院

出　　处：《中华实验外科杂志》2014年第7期1508-1510,共3页Chinese Journal of Experimental Surgery

基　　金：广东科技技术国际合作项目（A002013033）;吴阶平医学基金会临床科研专项基金资助项目（320.6750.12376）;广东省梅州市科技计划项目（20128104）

摘　　要：目的 探讨肝再生磷酸酶-3（PRL-3）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（5 mg/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。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 re- action （Real-time PCR） was used to screen the elevated mRNA of cytokines of the TAMs, and western blot- ting 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 sig- nificantly 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 cocuhure medium, the invasion of Lovo-P ceils 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.

关 键 词：肝再生磷酸酶-3 肿瘤相关性巨噬细胞 白细胞介素-6 结肠癌 

分 类 号：R735.35[医药卫生—肿瘤]