摘要
目的观察钙网蛋白(calreticulin,CRT)对内质网应激诱导的肝癌细胞转移和侵袭能力的影响。方法利用衣霉素刺激肝癌细胞SMMC-7721、HepG2建立内质网应激模型,用Western blot技术检测内质网应激指标免疫球蛋白结合蛋白(binding immunoglobulin protein,BIP)及钙网蛋白、MMP-9的表达,通过划痕实验、Transwell侵袭小室检测肝癌细胞转移和侵袭能力,并用小干扰RNA沉默钙网蛋白联合衣霉素处理后观察肝癌细胞转移、侵袭能力及MMP-9的表达。结果与阴性对照组相比,衣霉素处理肝癌细胞SMMC-772I及HepG2后BIP、细胞膜上钙网蛋白、MMP-9表达量增加,且转移和侵袭能力增加(P〈0.05);沉默钙网蛋白联合衣霉素处理后,与衣霉素单独处理相比,沉默钙网蛋白联合衣霉素组肝癌细胞SMMC-7721及HepG2MMP-9表达量降低,且转移和侵袭能力降低(P〈0.05)。结论内质网应激诱导钙网蛋白发生膜转位增强肝癌细胞的转移和侵袭能力。
Objective To explore the role of calreticulin (CRT) on endoplasmic reticulum stressinduced migration and invasion of SMMC-7721 and HepG2 hepatocellular carcinoma cells. Methods Endoplasmic retieulum stress was established in SMMC-7721 and HepG2 hepatoeellular carcinoma cells using ER stress inducer tunieamycin. Expressions of binding immunoglobulin protein (BIP), CRT, MMP-9 were examined by Western blot. The ability of migration and invasion of hepatocellular carcinoma ceils were detected by wound healing assay and transwell assay respectively. Using small interfering RNA and tunieamyein to observe its effect on the expression of MMP-9, migration and invasion in hepatocellular carcinoma cells. Results The expressions of BIP, CRT on the membrane, MMP-9, the wound healing rate and the numbers of invasion dramatically increased after tunieamyein incubation ( P 〈 0. 05 ). Data showed that expressions of MMP-9 and the wound healing rate and the numbers of invasion dramatically decreased after treatment with small interfering RNA (CRT) and tunieamyein( P 〈 0. 05). Conclusions Endoplasmic reticulum stress-induced calreticulin transloeation to cell surface promotes hepatocellular cancer ceils migration and invasion.
出处
《中华普通外科杂志》
CSCD
北大核心
2015年第10期804-807,共4页
Chinese Journal of General Surgery
基金
国家自然科学基金资助项目(U1204815)
河南省高校科技创新人才支持计划基金资助项目(14HASTIT030)
郑州大学青年教师专项科研基金资助项目(14214112090)
关键词
癌
肝细胞
内质网应激
钙网蛋白
Carcinoma, hepatoeellular
Endoplasmic retieulum stress
Calretieulin