摘要
目的探讨低氧诱导因子-1α（HIF-1α）在胆囊癌中的表达、临床意义及其在二甲双胍抑制胆囊癌GBC—SD细胞转移中的作用及机制。方法收集2016年6月至2017年2月在郑州大学第一附属医院接受外科手术的24例患者的胆囊癌组织和5例慢性胆囊炎组织，免疫组化及实时荧光定量（qPCR）检测胆囊癌、癌旁及慢性胆囊炎中HIF-1α表达，分析其临床意义；利用低氧环境培养诱导胆囊癌转移；分别采用划痕试验及Transwell侵袭小室检测细胞转移能力，应用蛋白印迹（Western blot）技术及免疫荧光检测细胞HIF-1α及VEGF表达量，应用2-甲氧基雌二醇（2-MeoE2）特异性抑制HIF-1α表达。结果胆囊癌组织中HIF-1α阳性表达率高于癌旁组织，差异有统计学意义（P〈0．05）。患者HIF-1α阳性率与胆囊癌的淋巴结转移、TNM分期相关（P〈0．05）。对照组（常氧）及低氧（1％ O2）组患者48h划痕修复率分别为（46．5±4．8）％、（67．3±4．0）％；对照组及低氧组患者24h穿膜细胞数分别为（147．4±11．7）个、（234．4±17．7）个。与对照组相比，低氧组胆囊癌GBC—SD细胞转移能力增加，HIF-1α及VEGF表达量增加（P〈0．05）。对照组、二甲双胍组、低氧组、二甲双胍联合低氧组48h划痕修复率分别为（40．6±7．1）％、（16．4±9．4）％、（69．5±4．0）％、（22．4±7．4）％；对照组、二甲双胍组、低氧组、二甲双胍联合低氧组24h穿膜细胞数分别为（148．4±6．9）个、（90．0±8．4）个、（185．8±10．2）个、（113．4±8．6）个。与低氧组比较，二甲双胍抑制低氧诱导胆囊癌细胞转移，同时HIF-1α及VEGF表达量下降（P〈0．05）。对照组、2-MeoE2组、低氧组、2-MeoE2联合低氧组48h划痕修复率分别为（43．4±4．4）％、（25．9±9．0）％、（63．3±2．2）％、（46．2±4．5）％。对照组、2-MeoE2组、低氧组、2-MeoE2�
Objective To study the expression and the clinical significance of hypoxia-induced factor-1α （HIF-1α） in gallbladder cancer tissues, and the role and mechanism of HIF-1α in metformin-suppressed metastasis in gallbladder carcinoma GBC-SD cells. Methods 24 specimens of gallbladder cancer tissues and 5 specimens of chronic cholecystitis were collected from the First Affiliated Hospital of Zhengzhou University between June 2016 and February 2017. Immunohistochemistry and qPCR were used to detect the expression of HIF-1α in gallbladder cancer tissues, in adjacent non-cancer tissues and in chronic cholecystitis, and the clinical significance was analyzed. The model of metastasis was induced by hypoxia; the wound healing assay and the Transwell assay were used to detect the ability of cell metastasis; the expressions of HIF-1α and VEGF in gallbladder carcinoma GBC-SD ceils were detected by western blotting assay and immunofluorescence. Results The expression of HIF-1α in gallbladder cancer tissues was higher than the adjacent non-cancer tissues and in chronic cholecystitis. The expression of HIF-1α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues （ P 〈0. 05）. The wound healing rate after 48 h in the negative control group and in the treatment with hypoxia group （ 1% O2 ） in GBC-SD ceils were （46. 5 ± 4. 8 ） % and （67.3 ± 4.0 ） %, respectively. The Transwell data showed that the numbers of metastasis after 24 h in the negative control group and in the treatment with hypoxia group GBC-SD cells were （ 147.4 ± 11.7 ） and （234.4 ±17.7 ）, respectively. When compared with the negative control group, treatment with hypoxia significantly increased the ability of metastasis and up-regnlated the expression of HIF-1α and VEGF in GBC-SD cells （P 〈 0.05 ）. The wound healing rate after 48 h in the negative control group, the metformin group, the hypoxia group and the metformin and hypoxia group in GBC-SD cells were （40. 6 ± 7. 1 ） %, （ 16. 4
出处
《中华肝胆外科杂志》
CSCD
北大核心
2017年第6期383-388,共6页
Chinese Journal of Hepatobiliary Surgery
基金
河南省高校科技创新人才支持计划（14HASTIT030）
郑州大学优秀青年教师发展基金（1421412090）
关键词
胆囊癌
低氧诱导因子-1Α
二甲双胍
低氧
转移
Gallbladder carcinoma
Hypoxia-induced factor-1α
Metformin
Hypoxia
Metastasis
