实用医学杂志 ›› 2022, Vol. 38 ›› Issue (19): 2414-2418.doi: 10.3969/j.issn.1006⁃5725.2022.19.007

• 基础研究 • 上一篇    下一篇

乙酰肝素酶抑制剂OGT2115通过Wnt/β⁃catenin 信号 通路促进胆囊癌细胞的凋亡 

陈佳伟1常卫才1刘子祥王郑林周少波1
  

  1. 1 蚌埠医学院第二附属医院普外科(安徽蚌埠233000);2 安徽省组织移植重点实验室(安徽蚌埠233000) 

  • 出版日期:2022-10-10 发布日期:2022-10-10
  • 通讯作者: 周少波 E⁃mail:zhoushaobodoctor@ sina.com
  • 基金资助:
    安徽省卫生健康委科研项目(编号:AHWJ2021a012);安徽省高等学校自然科学研究项目(编号:KJ2020A0564);蚌埠医学院自然科学重点项目(编号:2021byzd197)

Heparanase inhibitor OGT2115 promotes apoptosis in gallbladder carcinoma cells through the Wnt/β ⁃ catenin signaling pathway

CHEN Jiawei*,CHANG Weicai ,LIU Zixiang ,WANG Zhenglin ,ZHOU Shaobo.   

  1. The Second Affiliated Hospital of Bengbu Medical CollegeBengbu 233000China*Anhui Province Key Laboratory of Tissue TransplantationBengbu 233000China 
  • Online:2022-10-10 Published:2022-10-10
  • Contact: ZHOU Shaobo E⁃mail:zhoushaobodoctor@ sina.com

摘要:

目的 探讨乙酰肝素酶抑制剂 OGT2115Wnt/β⁃catenin 信号通路在对胆囊癌细胞生物学功能的影响。方法 用不同浓度梯度的 HPSE 抑制剂 OGT2115 作用于胆囊癌细胞 GBC⁃SDCCK⁃8 法检测 OGT2115 GBC⁃SD 细胞的抑制率,流式细胞术检测 GBC⁃SD 的凋亡比例,ELISA 检测 OGT2115 对细胞 上清液 HPSEFas 以及 β⁃catenin 活性,将实验分为对照组、OGT2115 组和 IWR⁃1+OGT2115 组,qRT⁃PCR Western blot 检测 OGT2115 对细胞凋亡基因(Caspase⁃3BaxBcl⁃2)和 Wnt/β⁃catenin 信号通路相关基因 (β⁃cateninc⁃MYCCyclinD1E⁃cadherin)表达的影响。结果 OGT2115可以抑制GBC⁃SD增殖,且具有浓度依 赖性。OGT2115 抑制乙酰肝素酶活性,促进细胞凋亡。qRT⁃PCR Western blot 结果显示,OGT2115 促进 caspase⁃3cleaved⁃caspase⁃3Bax 基因表达水平,而 Bcl⁃2、信号通路相关基因 β⁃cateninc⁃MYCCyclin D1E⁃cadherin表达水平下降,且IWR⁃1抑制该信号通路后,OGT2115对凋亡及信号通路的作用更加明显。结论 乙酰肝素酶抑制剂 OGT2115 抑制 GBC⁃SD 细胞的生长与活性,可能通过 Wnt/β⁃catenin 通路促进凋亡。

关键词:

胆囊癌, 乙酰肝素酶, 肝素酶抑制剂, 细胞凋亡, 信号通路

Abstract:

Objective To investigate the effect of acetyl heparanase inhibitor OGT2115 and Wnt/β⁃catenin signaling pathway on the biological function of gallbladder carcinoma cells. Methods GBC⁃SD cells were treated with different gradients of HPSE inhibitor OGT2115,the inhibition of GBC⁃SD cells was detected by CCK⁃8 assay, and the apoptotic ratio of GBC⁃SD cells was detected by flow cytometry. The detection of OGT2115 on HPSE,Fas and β ⁃ catenin activities in cell supernatants was performed by ELISA. The experiment was divided into control group,OGT2115 group and IWR⁃1+OGT2115 group,the effects of OGT2115 on the expression of apoptotic genes (Caspase⁃3,Bax,Bcl⁃2)and Wnt/β⁃catenin signaling pathway⁃related genes(β⁃catenin,c⁃MYC,CyclinD1, E⁃cadherin)by qRT⁃PCR and Western blot. Results OGT2115 inhibited the proliferation of GBC⁃SD in a concen⁃ tration ⁃ dependent manner. OGT2115 inhibited acetyl heparanase activity and promoted apoptosis. The qRT ⁃ PCR and Western⁃Blot results showed that OGT2115 promoted the expression levels of caspase⁃3,cleaved⁃caspase⁃3, and Bax genes,while the expression levels of Bcl⁃2,signaling pathway⁃related genes β⁃catenin,c⁃MYC,Cyclin D1,and E⁃cadherin decreased,and the effect of OGT2115 on apoptosis and signaling pathway was more obvious after IWR ⁃ 1 inhibited this signaling pathway. Conclusions The acetyl heparinase inhibitor OGT2115 inhibited the growth and activity of GBC⁃SD cells and may promote apoptosis through the Wnt/β⁃catenin signaling pathway.

Key words:

gallbladder carcinoma, heparanase, heparanase inhibitor, apoptosis, signal pathway