TW-37调节NF—kB信号通路抑制体外胰腺癌细胞转移的实验研究  

TW-37 inhibited metastasis in pancreatic cancer via regulating NF-kB signal in vitro

作　　者：吴隆超 王琳娜 刘瑞东[3] 王晓丽[3] 田文霞[3] 李兴涛[3] 张俊[4] 

机构地区：[1]蓬莱市人民医院普外科,山东蓬莱265600 [2]山东省青岛疗养院检验科 [3]莱芜市人民医院检验科 [4]浙江省人民医院肿瘤外科

出　　处：《中华胰腺病杂志》2016年第4期237-242,共6页Chinese Journal of Pancreatology

摘　　要：目的观察TW-37干预对胰腺癌细胞增殖、凋亡、侵袭及血管生成的影响，探讨其可能机制。方法应用TW-37干预胰腺癌细胞株BxPC3和HPAC，以转染NF-KBp65eDNA（p65eDNA）、靶向NF—KBp65的siRNA（siRNA—p65）细胞及未干预细胞作为对照组。采用MTT和ELISA法检测细胞增殖和细胞凋亡；应用Transwell小室检测细胞侵袭能力；采用体外人脐静脉内皮细胞（HUVECs）成管实验观察细胞培养上清对血管生成的影响；ELISA法检测细胞NF．KB活性；蛋白质印迹法检测细胞NF-kB靶向调节蛋白VEGF、MMPO表达。结果TW-37呈浓度和时间依赖性抑制BxPC3、HPAC细胞的增殖，诱导两株细胞的凋亡（A405值1．29±0．21比0．09±0．01，1．07±0．18比0．08±0．01），抑制细胞NF．KB活性及NF-kB p65、VEGF、MMPO蛋白表达，差异均有统计学意义（P值均〈0．05）。对照组、0．75μmoVLTW．37干预组的穿膜细胞数分别为（46．7±5．24）、（10．3±1．26）＋／200倍视野；HUVECs形成的血管数分别为（39．4±4．36）、（7．84±1．25）＋／200倍视野，差异均有统计学意义（P值均为0．001）。转染p65eDNA的两株细胞的NF—KB活性较对照组显著增加，用TW-37干预后NF—kB活性下降；转染siRNA．p65的两株细胞的NF-kB活性较对照组显著下降，用TW-37干预后NF—kB活性进一步下降，差异均有统计学意义（P〈0．05或〈0．01）。转染p65eDNA对两株细胞凋亡无明显影响，用TW-37干预后凋亡略有变化；转染siRNA—p65的两株细胞凋亡显著增加，用TW-37干预后又进一步增加，差异均有统计学意义（P值均〈0．01）。结论TW-37通过NF—kB抑制胰腺癌细胞增殖、侵袭和血管生成，诱导细胞凋亡。Objective To study the effect and mechanisms of TW-37 on cell proliferation, apoptosis, invasion and angiogenesis in pancreatic cancer cells in vitro and further explore the potential mechanism. Methods BxPC3 and HPAC cells were pretreated with TW-37 using untransfected or transfected with NF-kB 1365 eDNA（p65 eDNA）or NF-kB p65 siRNA（ siRNA-p65 ）cells as controls. Cell viability was determined by MTT assay. Cell apoptosis was assessed by enzyme-linked immunosorbent assay （ELISA）. Cell invasion and angiogenesis was detected by Transwell and endothelial tube formation assay of HUVECs. ELISA assay was used to measure the activity of NF-kB, and its target proteins of MMP-9 and VEGF were detected by western blot. Results TW-37 suppressed cell growth and induced apoptosis （ A405 ： 1.29 ± 0.21 vs 0. 09 ± 0.01, 1.07 ± 0.18 vs 0. 08 ± 0.01 ）, inhibited NF-kB activity and protein expression of NF-KB p65, VEGF and MMP-9（ all P 〈 0.05 ）in a dose- and time-dependent manner. The number of cells that invaded across the matrigel in the transwell chamber was （46.7 ± 5.24） and （ 10.3 ± 1.26 ） / x 200 in BxPC3 control and 0.75 μmol/L TW-37 group （ P = 0.001 ）. The number of tube formation was （ 39.4± 4.36 ） and （ 7.84 ± 1.25 ） / x 200, （ P = 0.001 ）. NF-kB activity was increased by p65 cDNA transfection, and decreased by TW-37 treatment in both of the two cell lines （P 〈 0.05 ）. However, NF-kB activity was deereased by p65 siRNA transfection, and greatly decreased by TW-37 treatment in both two cell lines （P 〈0.05 or P 〈0.01 ）. Transfection of p65 cDNA did not siguifieantly affect eell apoptosis. Transfeetion of p65 siRNA increased cell apoptosis, and greatly increased by TW-37 treatment in both two cell lines （all P 〈 0.01 ）. Conclusions TW-37 eould inhibit the proliferation, invasion and angiogenesis in panereatie cancer eells by regulating NF-kB signal pathway.

关 键 词：胰腺肿瘤 生长 肿瘤转移 病理性血管生成 TW-37 

分 类 号：R735.9[医药卫生—肿瘤]