奥巴克拉联合5-氟尿嘧啶对低氧性胰腺癌细胞上皮间质转化的影响

Effects of obatoclax plus 5-Fluorouracil on the epithelial mesenchymal transition of pancreatic cancer cells under hypoxia

目的:研究在低氧条件下,奥巴克拉(obatoclax)联合5-氟尿嘧啶(5-Fluorouracil,5-FU)对胰腺癌细胞株Bx PC-3细胞上皮间质转化(epithelial mesenchymal transition,EMT)的影响。方法:细胞分组为低氧组、5-FU组、obatoclax+5-FU组,利用MTT法检测各组Bx PC-3细胞增殖活性;利用实时荧光定量PCR(RT-q PCR)与Western blot检测各组Bx PC-3细胞中EMT相关基因与蛋白的表达情况。结果:在低氧条件下,obatoclax联合5-FU可抑制Bx PC-3细胞的增殖活性,且具有时间依赖性;obatoclax联合5-FU可进一步促进EMT的管家基因,转录因子Snail与Slug的表达升高;obatoclax联合5-FU促进上皮标记E-Cadherin的表达升高,及间质标记N-Cadherin、Fibronectin、Collagen Ⅰ、Vimentin的表达下降。结论:在低氧状态下obatoclax联合5-FU可抑制胰腺癌Bx PC-3细胞的增殖活性,可通过抑制转录因子Snail与Slug的表达,上调上皮标记E-Cadherin的表达,下调间质标记N-Cadherin、Fibronectin、Collagen Ⅰ及Vimentin的表达,从而抑制肿瘤细胞上皮向间质的转化,进而降低肿瘤细胞的迁移、侵袭能力。

AIM： To explore the effect of obato- clax combined with 5-Fluorouracil （5-FU） in epithelial mesenchymal transition （EMT） of pancreatic cancer BxPC-3 cells and its possible mechanism. METHODS： MTT assay was performed to detect cell viability in hypoxic group, 5-FU group and obatoclax combined with 5-FU group. RT-qPCR and Western blotting were used to detect EMT-related genes and proteins expression in BxPC-3 cells of hypoxic group, 5-FU group and obatoclax combined with 5-FU group. RESULTS ： Under hypoxia condi- tion, obatoclax combined with 5-FU time-dependent- ly inhibited BxPC-3 cell proliferation activity. Obatoclax combined with 5-FU further down-regulated the expression of transcription factors Snail and Slug, as well as mesenchymal markers N-Cadherin,Fibronectin, Collagen I and Vimentin. Obatoclax combined with 5-FU up-regulated the expression of epithelial marker E-Cadherin. CONCLUSION： Under hypoxia condition, obatoclax combined with 5-FU inhibit the viability of BxPC-3 cells. Obatoclax combined with 5-FU can up-regulate E-Cadherin and down-regulate N-Cadherin, Fibronectin, Collagen I and Vimentin through inhibiting Snail and Slug, thus inhibiting EMT, reducing migration and invasion ability of BxPC-3 cells.