肝再生磷酸酶-3上调转化生长因子-beta促进结肠癌LoVo细胞侵袭的机制研究  

Phosphatase of regenerating liver-3 promoted the invasion of LoVo cells through TGF-beta

作　　者：施景龙 罗兴喜[2] 许鹤洋[2] 蓝球生 黄永亮[2] 褚忠华[2] 

机构地区：[1]广州市南沙区第一人民医院普外科,广州511440 [2]中山大学孙逸仙纪念医院胃肠外科,广州510120

出　　处：《岭南现代临床外科》2017年第3期287-289,294,共4页Lingnan Modern Clinics in Surgery

基　　金：广东省科技计对外科技合作项目(2013B051000025;2015A050502021);广东省医学科研基金(B2014133);广东省自然科学基金(2014A030313054)

摘　　要：目的探讨转化生长因子-beta(TGF-beta)通路在肝再生磷酸酶-3(PRL-3)促进结肠癌细胞侵袭和转移中的作用机制。方法酶联免疫吸附试验ELISA检测实验组与对照组结肠癌细胞TGF-beta水平表达差异,Transwell侵袭小室法检测Lo Vo细胞侵袭能力变化,Western blot检测相关通路蛋白水平变化。结果 ELISA实验检测结肠癌细胞转染PRL-3后,稳定表达PRL-3的Lo Vo细胞(Lo Vo-P)与对照组(Lo Vo-C)相比,TGF-beta表达明显升高(114±11 pg/m L比56±8 pg/m L,P<0.01)。Transwell侵袭实验提示不同浓度的TGF-beta中和抗体(1、10、100 ng/m L)作用Lo Vo-P后其侵袭能力逐渐降低,对照组穿膜细胞数为142.7±11.3个,实验组分别为96.1±8.2、67.3±9.4、48.6±6.4个(P<0.05)。蛋白印记实验提示PI3K/AKT信号通路参与了其诱导侵袭的过程,使用PI3K抑制剂LY294002(10 mg/m L)后,Lo Vo-P的AKT磷酸化蛋白的表达水平降低2.5倍(P<0.05)。结论 PRL-3能诱导结肠癌Lo Vo细胞分泌TGF-beta,激活PI3K/AKT信号通路进而促进结肠癌Lo Vo细胞侵袭。Objective To investigate role of TGF-beta in promoting the colorectal cancer Lo Vo cells invasion induced by phosphatase of regenerating liver-3(PRL-3). Methods ELISA assay was to used to detect the level of TGF-beta in Lo Vo-P and Lo Vo-C. Invasion assays were applied to determine the effect of TGF-beta on the ability of PRL-3. Western blot was used to detect the proteins of p-AKT and AKT. Results Elisa assay displayed that the protein level of TGF-beta of Lo Vo-P was higher than Lo Vo-C(114±11 pg/ml vs. 56±8 pg/ml,P<0.01). When added different dosages of neutralizing antibody of TGF-beta(1,10,00 ng/ml)in culture medium of Lo Vo-P,the invasion were decreased significantly(96.1±8.2,67.3±9.4 and 48.6±6.4,respectively)and all Pvaluesless than 0.05). Western blot reminded the proteins of p-AKT in Lo Vo-P were higher than Lo Vo-C,and the expression of p-AKT in Lo Vo-P was decreased by 2.5-fold after treating Lo Vo-P with the PI3 K inhibitor LY294002(10 mg/m L)(P<0.05). Conclusion PRL-3 could up-regulate the expression of TGF-beta in colorectal cancer Lo Vo cells and promote the invasion of Lo Vo cells,in which PI3K/AKT signaling pathway may be involved.

关 键 词：肝再生磷酸酶-3 结肠癌 转化生长因子-beta 侵袭 

分 类 号：R73-37[医药卫生—肿瘤]