中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1908-1910
,共3页
黄伟华%来伟%蓝球生%曾献清%张旸%褚忠华
黃偉華%來偉%藍毬生%曾獻清%張旸%褚忠華
황위화%래위%람구생%증헌청%장양%저충화
肝再生磷酸酶-3%结肠癌%肿瘤相关性巨噬细胞%迁移
肝再生燐痠酶-3%結腸癌%腫瘤相關性巨噬細胞%遷移
간재생린산매-3%결장암%종류상관성거서세포%천이
Phosphatase of regenerating liver-3%Colon cancer%Tumor-associated macrophages%Migration
目的 观察肝再生磷酸酶-3(PRL-3)促进结肠癌细胞LoVo分泌趋化因子26(CCL26),对肿瘤相关性巨噬细胞的趋化、聚集作用.方法 实时定量反转录聚合酶链反应(RT-qPCR)检测CCL26 mRNA水平表达差异;酶联免疫吸附试验(ELISA)检测CCL26蛋白水平表达差异,检测添加核因子κB(NF-κB)抑制剂BAY 11-7082(5、10、15 mol/L)后,CCL26蛋白水平;Transwell迁移实验检测LoVo-P、LoVo-C与TAM共培养24h后,对TAM迁移作用.结果 RT-qPCR检测结果显示,LoVo-P对比LoVo-C,CCL26升高(46±5)倍,ELISA检测CCL26蛋白水平,LoVo-P为(91±5) ng/L,LoVo-C为(61 ±3) ng/L,LoVo-P加入核因子(NF)-κB抑制剂BAY 11-7082(5、10、15 mol/L)后,CCL26蛋白水平呈浓度梯度降低(P<0.01);Transwell迁移实验结果显示,LoVo-P对比LoVo-C明显对TAM有趋化作用,加入NF-κB抑制剂BAY 11-7082后,迁移作用明显减弱,不同浓度的CCL26(1、10、100 μg/L),能对TAM的趋化作用呈浓度梯度增高(P<0.01).结论 PRL-3能促进结肠癌LoVo细胞分泌CCL26,通过CCL26可以诱导TAM趋化、聚集.
目的 觀察肝再生燐痠酶-3(PRL-3)促進結腸癌細胞LoVo分泌趨化因子26(CCL26),對腫瘤相關性巨噬細胞的趨化、聚集作用.方法 實時定量反轉錄聚閤酶鏈反應(RT-qPCR)檢測CCL26 mRNA水平錶達差異;酶聯免疫吸附試驗(ELISA)檢測CCL26蛋白水平錶達差異,檢測添加覈因子κB(NF-κB)抑製劑BAY 11-7082(5、10、15 mol/L)後,CCL26蛋白水平;Transwell遷移實驗檢測LoVo-P、LoVo-C與TAM共培養24h後,對TAM遷移作用.結果 RT-qPCR檢測結果顯示,LoVo-P對比LoVo-C,CCL26升高(46±5)倍,ELISA檢測CCL26蛋白水平,LoVo-P為(91±5) ng/L,LoVo-C為(61 ±3) ng/L,LoVo-P加入覈因子(NF)-κB抑製劑BAY 11-7082(5、10、15 mol/L)後,CCL26蛋白水平呈濃度梯度降低(P<0.01);Transwell遷移實驗結果顯示,LoVo-P對比LoVo-C明顯對TAM有趨化作用,加入NF-κB抑製劑BAY 11-7082後,遷移作用明顯減弱,不同濃度的CCL26(1、10、100 μg/L),能對TAM的趨化作用呈濃度梯度增高(P<0.01).結論 PRL-3能促進結腸癌LoVo細胞分泌CCL26,通過CCL26可以誘導TAM趨化、聚集.
목적 관찰간재생린산매-3(PRL-3)촉진결장암세포LoVo분비추화인자26(CCL26),대종류상관성거서세포적추화、취집작용.방법 실시정량반전록취합매련반응(RT-qPCR)검측CCL26 mRNA수평표체차이;매련면역흡부시험(ELISA)검측CCL26단백수평표체차이,검측첨가핵인자κB(NF-κB)억제제BAY 11-7082(5、10、15 mol/L)후,CCL26단백수평;Transwell천이실험검측LoVo-P、LoVo-C여TAM공배양24h후,대TAM천이작용.결과 RT-qPCR검측결과현시,LoVo-P대비LoVo-C,CCL26승고(46±5)배,ELISA검측CCL26단백수평,LoVo-P위(91±5) ng/L,LoVo-C위(61 ±3) ng/L,LoVo-P가입핵인자(NF)-κB억제제BAY 11-7082(5、10、15 mol/L)후,CCL26단백수평정농도제도강저(P<0.01);Transwell천이실험결과현시,LoVo-P대비LoVo-C명현대TAM유추화작용,가입NF-κB억제제BAY 11-7082후,천이작용명현감약,불동농도적CCL26(1、10、100 μg/L),능대TAM적추화작용정농도제도증고(P<0.01).결론 PRL-3능촉진결장암LoVo세포분비CCL26,통과CCL26가이유도TAM추화、취집.
Objective To investigate the phosphatase of regenerating liver-3 (PRL-3) promoting colon cancer cells LoVo to secrete chemokine 26 (CCL26),which enhances tumor-associated macrophages (TAMs) migration.Methods We used real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR),and enzyme linked immunosorbent assay (ELISA) to detect the mRNA and protein expression of CCL26,and the protein expression of CCL26 after treatment of LoVo cells with nuclear factor-κB (NF-κB) inhibitor,BAY 11-7082 (5,10,and 15 mol/L).Invasion assays were applied to determine the effect of CCL26 on the ability of PRL-3 promoting migration of TAMs.Results RT-qPCR showed that the CCL26 mRNA expression of LoVo-P was higher than LoVo-C [(46 ± 5) fold].ELISA displayed that the protein level of CCL26 of LoVo-P was higher than LoVo-C,and it was attenuated in a concentration-dependent manner by treating LoVo-P with the NF-κB inhibitor,BAY 11-7082 (5,10,and 15 mol/L).Transwell invasion assays showed that the migration of TAMs was enhanced when TAMs were cocultured with LoVo-P cells,and it can be inhibited by NF-κB inhibitor,BAY 11-7082.Also the migration of TAMs was enhanced in a concentration-dependent manner when TAMs were cocultured with different concentrations of CCL26 (P < 0.01).Conclusion PRL-3 promotes colon cancer cells to secrete chemokine CCL26,which can enhance TAMs migration.