肝癌患者来源异种移植瘤模型的建立及药物敏感性研究  

Establishment of liver cancer patient-derived xenograft model and exploration of drug sensitivity

作　　者：刁文婧 吴亚菲 武传星 陈大红 杨蕙华 毛鹏娟 李琴[1] Diao Wenjing;Wu Yafei;Wu Chuanxing;Chen Dahong;Yang Huihua;Mao Pengjuan;Li Qin(Department of Clinical Pharmacy,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China)

机构地区：[1]上海交通大学医学院附属第一人民医院临床药学科,上海200080 [2]上海交通大学医学院附属第一人民医院肝胆外科,上海200080

出　　处：《中华实验外科杂志》2023年第12期2576-2579,共4页Chinese Journal of Experimental Surgery

基　　金：上海市申康第二轮临床三年行动计划-临床研究青年项目(SHDC2020CR4076)。

摘　　要：目的通过建立肝癌患者来源的异种移植瘤模型(PDX)探究肝癌索拉菲尼敏感性相关的长链非编码RNA(lncRNA)。方法选取上海市第一人民医院2021年5月至2022年10月的17例肝癌标本建立PDX模型。PDX模型传代14 d后通过随机数表法分为对照组与索拉菲尼给药组。给药21 d,观察7 d后收取肿瘤组织,给药期间记录小鼠体重。根据公式:肿瘤生长抑制(TGI)指数=ΔT/ΔC,计算PDX肿瘤组织TGI指数并评价药物敏感性,通过lncRNA测序筛选肝内胆管癌(ICC)索拉菲尼敏感与耐药的差异lncRNA。在人ICC细胞系HuCCT1中转染小干扰RNA(siRNA)降低lncRNA SCDAL的表达,根据siRNA序列分组为ctr、si-1、si-2和si-3。采用细胞计数试剂盒(CCK-8)检测细胞增殖能力,根据给药浓度分组为0μmol/L(0.1%二甲基亚砜)、4μmol/L、10μmol/L。组间比较采用t检验,多组间两两比较采用单因素方差分析,Turkey事后检验确定组间差异。结果肝细胞肝癌(HCC)及ICC PDX模型鼠索拉非尼给药组与对照组体重差异无统计学意义[HCC:(24.72±2.21)g比(25.80±2.30)g,t=0.486,P>0.05;ICC:(24.87±4.30)g比(23.11±2.78)g,t=0.478,P>0.05];lncRNA SCDAL在PDX耐药样本中表达显著低于敏感组(|log2FoldChange|=Inf,P<0.05);Si-3敲减组lncRNA SCDAL相对表达量显著低于对照组(0.586±0.080比1.000±0.234,P<0.05)。索拉菲尼给药条件下敲减组细胞吸光度值显著高于对照组(4μmol/L:1.11±0.06比0.86±0.07,t=4.635,P<0.01;10μmol/L:0.52±0.03比0.42±0.02,t=4.389,P<0.05)。结论通过构建肝癌患者PDX模型并测序验证发现lncRNA SCDAL表达与ICC索拉菲尼敏感性呈正相关。Objective Explore sorafenib sensitivity-related long non-coding RNA(lncRNA)in liver cancer through establishing liver cancer patient-derived xenograft(PDX)model.Methods Select 17 cases of liver cancer samples from May 2021 to October 2022 in Shanghai General hospital to establish PDX models.PDX models were randomly divided into control group and sorafenib administration group by random number method.after passaging 14 d.Tumor tissues were collected after 21 d of sorafenib administration and 7d of observation,mice weights were recorded.Drug susceptibility of PDX tumor tissues was evaluated by calculating Tumor Growth Inhibition(TGI)value according to formula:TGI=ΔT/ΔC.LncRNA sequencing was used to screen different lncRNA between sorafenib resistant and sensitive tissues in intrahepatic cholangiocarcinoma(ICC).Knockdown of lncRNA SCDAL in human ICC cell line HuCCT1 was performed by transfecting small interfering RNA(siRNA),which were grouped into ctr,si-1,si-2 and si-3 according to siRNA sequences.Cell counting kit-8(CCK-8)assay was applied to detect cell proliferation ability,which were grouped into 0μmol/L(0.1%dimethyl sulfoxide),4μmol/L and 10μmol/L according to drug concentration.Results There was no significant difference in body weight of hepatocellular carcinoma(HCC)and ICC PDX mice between sorafenib administration group and control group[HCC:(24.72±2.21)g vs.(25.80±2.30)g,t=0.486,P>0.05;ICC:(24.87±4.30)g vs.(23.11±2.78)g,t=0.478,P>0.05].The expression of lncRNA SCDAL in PDX drug-resistant samples was significantly lower than drug-sensitive group(|log2FoldChange|=Inf,P<0.05).Relative expression of lncRNA SCDAL in knockdown group of si-3 was significantly lower than control group(0.586±0.080 vs.1.000±0.234,P<0.05).Knockdown group possessed significant higher cell absorbance value than control group under sorafenib administration(4μmol/L:1.11±0.06 vs.0.86±0.07,t=4.635,P<0.01;10μmol/L:0.52±0.03 vs.0.42±0.02,t=4.389,P<0.05).Conclusion The present study found that lncRNA SCDAL expression was posi

关 键 词：患者来源的异种移植瘤模型 肝癌 索拉菲尼 长链非编码RNA 肝内胆管癌 

分 类 号：R735.7[医药卫生—肿瘤]