肝细胞癌索拉非尼耐药细胞株鸡胚种植瘤模型的建立  

Establishment of xenografts in chicken chorioallantoic membrane model with hepatocellular carcinoma sorafenib resistant cell line

作　　者：张磐石 张菁 刘东伯 孙伟 刘顺芳 Zhang Panshi;Zhang Jing;Liu Dongbo;Sun Wei;Liu Shunfang(Department of Thyroid and Breast Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Department of Oncology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院甲状腺乳腺外科,武汉430030 [2]华中科技大学同济医学院附属同济医院肿瘤科,武汉430030

出　　处：《中华实验外科杂志》2021年第10期2034-2037,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81602626);华中科技大学自主创新基金资助项目(2016YXMS111);湖北省自然科学基金(2019CFB650);湖北陈孝平科技发展基金会肝胆胰恶性肿瘤研究基金(CXPJJH12000001-2020308)。

摘　　要：目的探讨建立肝细胞癌索拉非尼耐药细胞株鸡胚种植瘤模型的可行性。方法采用药物连续诱导法,构建肝癌索拉非尼耐药细胞株;利用鸡胚动物模型,建立索拉非尼耐药细胞株的移植瘤;初步探讨耐药细胞株鸡胚种植瘤模型成瘤特点,分析相关靶点和信号通路的表达。采用t检验。结果筛选到索拉非尼耐药的HepG2细胞(sHepG2)半数抑制浓度(IC50)为(88.7±7.6) μmol/ml,显著高于HepG2(t=16.416,P<0.01);建立HepG2与sHepG2细胞株鸡胚种植瘤模型各5例,成功种植成瘤率为70%[(3+4)/(5+5)];种植瘤肿瘤鲜重HepG2组[(0.093 2±0.012 2) g]低于sHepG2组[(0.141 1±0.019 4) g,t=3.709,P<0.05];成瘤组织蛋白质印迹法(Western blot)检测,sHepG2组磷脂酰肌醇-3激酶(PI3K)(t=9.336,P<0.01)、磷酸化蛋白激酶(p-Akt/p-PKB)(t=7.123,P<0.01)表达显著高于HepG2组,人第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)(t=6.164,P<0.01)、成纤维细胞生长因子受体1 (FGFR1)(t=6.297,P<0.01)表达显著低于HepG2组。结论体外建立索拉非尼诱导的HepG2耐药细胞株,并在鸡胚移植瘤动物模型中成瘤,移植瘤生长以及瘤组织相关靶点与信号通路的差异性表明,鸡胚种植瘤模型可作为肝细胞癌索拉非尼耐药的在体研究方法。Objective To establish the xenografts in chicken chorioallantonic membrane(CAM)model with sorafenib-induced drug-resistant hepatocellular carcinoma cell lines.Methods Applying continuity induction in vitro to obtain hepatocellular carcinoma sorafenib resistant cell lines.Using chicken CAM model to gain xenografts tumors in vivo.Exploring the characteristics of xenografts tumors,analyzing the expression of related target genes and signaling pathway included.For statistical analyses,t-test was used,and the results were considered significant at P<0.05.Results Sorafenib-resistant HepG2 cell line was screened,named sHepG2,which half maximal inhibitory concentration(IC50)[(88.7±7.6)μmol/ml]was higher than that of HepG2 significantly(t=16.416,P<0.01).5 cases of chick CAM models of HepG2 and sHepG2 cells were established respectively,and the xenograft tumor formation rate was 70%[(3+4)/(5+5)].The fresh weight of xenograft tumors in HepG2 group[(0.0932±0.0122)g]was significantly lighter than that in sHepG2 group(t=3.709,P<0.05).ComparedsHepG2 with HepG2,the expression of phosphoinositide 3-kinase(PI3K)(t=9.336,P<0.01)and phospho-protein kinase B(p-Akt)(t=7.123,P<0.01)werehigher,while phosphatase and tensin homolog deleted on chromosome ten(PTEN)(t=6.164,P<0.01)and fibroblast growth factor receptor 1(FGFR1)(t=6.297,P<0.01),lowersignificantly.Conclusion The establishment of sorafenib-induced HepG2 drug-resistant cell lines in vitro,the constructing xenografts in chicken CAM model with HepG2 drug-resistant cells,the exploration growth of xenografts tumors and relative protein expressions indicate that the chicken CAM model could be useful for HCC sorafenib-resistant research in vivo.

关 键 词：肝细胞癌 索拉非尼 鸡胚模型 种植瘤 药物耐药 

分 类 号：R735.7[医药卫生—肿瘤] R-332[医药卫生—临床医学]