白细胞介素-1β对胆囊癌细胞生长和迁移的影响

Effects of IL-1 β on proliferation and migration of gallbladder cancer cells

目的探讨白细胞介素-1β（IL-1β）对胆囊癌细胞生长、迁移和侵袭的影响。方法ELISA法检测胆囊癌、慢性胆囊炎组织、正常胆囊组织标本以及细胞培养基中IL-1β的含量；实时-PCR技术检测IL-1β在胆囊癌细胞株（GBC-SD和SGC996）和肝内胆管上皮细胞株HIBEpic中的表达水平；采用WST-1法和动物模型检测IL-1β在体内和体外对胆囊癌细胞生长的影响；采用Transwell试验检测胆囊癌细胞迁移能力改变；实时-PCR和蛋白印迹法检测IL-1β作用后Twist基因表达水平，并在沉默Twist基因表达后进一步检测IL-1β对细胞生长和迁移的作用。结果IL-1β在胆囊癌和慢性胆囊炎组织标本中含量[（616．4±95．7）pg／ml和（422．3±48．9）pg／ml]远远高于胆囊正常组织[（66．4±35．0）pg／ml，P〈0．05]；IL-1β在GBC-SD和SGC996细胞培养液中的含量[（587．4±99．8）pg／ml和（657．2±76．6）pg／ml]高于HIBEpic[（38．4±12．1）pg／ml，P〈0．05]；加入外源性重组IL-1β可以显著促进GBC-SD和SGC996细胞的生长和迁移（P〈0．05）；外源性IL-1β可以诱导Twist表达升高。沉默Twist表达，外源性IL-1β无法促进胆囊癌细胞生长和迁移。结论IL-1β通过诱导Twist基因表达促进胆囊癌细胞生长和转移。

Objective To investigate the effects of -1β on proliferation and migration of gallbladder cancer cells. Methods The secretion of -1β in tissues of gallbladder cancer, chronic cholecystitis and normal gallbladder as well as in supernatant of gallbladder cancer cell lines （ GBC-SD, SGC996） and HIBEpic cells was determined by enzyme-linked immunosorbent assay （ELISA） method. The levels of -1β mRNA in GBC-SD, SGC996 and HIBEpic cells were measured by RT-PCR assay. The effects of exogenous IL-1β on the proliferation of GBC-SD and SGC996 cells in vitro and in vivo were evaluated using WST-1 assay and xenograft tumor model, respectively. The effects of exogenous -1β on the migration of GBC-SD and SGC996 cells in vitro were measured by Tranwell assay. The levels of Twist protein in GBC-SD and SGC996 cells were examined by western blot assay after treatment with exogenous IL-1 β. In addition, the proliferation and migration of GBC-SD and SGC996 cells after gene silencing of Twist by Twist-siRNA were also evaluated. Results The level of -1β protein in normal gallbladder was low （ 66.4 ± 35.0） pg/ml, while it was significantly increased in gallbladder cancer and chronic cholecystitis [ （616. 4 ± 95. 7 ） pg/ml and （422.3 ±48.9）pg/ml, P 〈 0.05 ]. The levels of IL-1β in GBC-SD and SGC996 cell culture medium [ （ 587.4 ± 99.8 ） pg/ml and （ 657.2 ±76.6 ） pg/ml ] were much higher than those in the HIBEpic cells [ （38.4 ± 12.1 ）pg/ml, P 〈 0.05 ]. Exogenous -1β promoted the proliferation of GBC-SD and SGC996 cells both in vitro and in vivo as well as migration in vitro （ P 〈 0.05 ）. The level of Twist protein was significantly increased after treatment with exogenous IL-1 β. In addition, gene silencing of Twist blocked IL-1 β- induced proliferation and migration of GBC-SD and SGC996 cells. Conclusion IL-1β promoted proliferation and migration of gallbladder cancer cells via Twist activation.