蒙药乌力地格调控PI3K/AKT/mTOR信号通路抑制肝门部胆管癌裸鼠移植瘤的机制研究  

To study the mechanism of Mongolian medicine Ulidig on inhibiting hilar cholangiocarcinoma xenografts in nude mice by regulating PI3K/AKT/mTOR signaling pathway

作　　者：胡向贇 贾剑超 戚晓辉 王石 HU Xiangyun;JIA Jianchao;QI Xiaohui;WANG Shi(Inner Mongolia Autonomous Region People's Hospital Department of Emergency Medicine,Emergency Medicine Center,Hohhot 010110,China;Inner Mongolia Autonomous Region People's Hospital Clinical Research Centera,Hohhot 010110,China;Inner Mongolia Autonomous Region People's Hospital Department of Hepatobiliary Pancreatic and Spleen Surgery,Hohhot 010110,China)

机构地区：[1]内蒙古自治区人民医院急诊医学中心急诊医学科,内蒙古呼和浩特010110 [2]内蒙古自治区人民医院临床医学研究中心,内蒙古呼和浩特010110 [3]内蒙古自治区人民医院肝胆胰脾外科,内蒙古呼和浩特010110

出　　处：《中国实验诊断学》2025年第2期189-196,共8页Chinese Journal of Laboratory Diagnosis

基　　金：内蒙古自治区自然科学基金项目(2019MS08081、2023MS08016);内蒙古自治区卫生健康科技计划项目(202201009);内蒙古医科大学联合项目(YKD2023LH080)。

摘　　要：目的探索蒙药乌力地格对肝门部胆管癌细胞株QBC939荷瘤裸鼠的作用效果并同时研究其作用效果与PI3K/AKT/mTOR信号通路的关系。方法用QBC939细胞在裸鼠皮下造模成功后,随机将荷瘤裸鼠分为吉西他滨组、生理盐水空白对照组、二甲基亚砜(Dimethy sulfoxide,DMSO)溶剂对照组、乌力地格组。按照分组情况给药,并且记录各组移植瘤体积及荷瘤裸鼠重量变化。连续给药15天,眼球采血后处死荷瘤裸鼠。用酶联免疫吸附试验(ELISA)检测血清中肿瘤标记物CEA、CA19-9的含量。取部分荷瘤裸鼠肿瘤组织进行HE染色及免疫组织化学法观察肿瘤细胞的染色情况、结构形态、坏死程度,以及PI3K、AKT、mTOR蛋白的表达情况;其余肿瘤组织通过qRT-PCR测定PI3K、AKT、mTOR的RNA相对表达量。结果(1)通过测量各个组移植瘤体积,发现与生理盐水组和DMSO组相比,乌力地格组肿瘤体积较小,差异有统计学意义(P<0.05);乌力地格组与吉西他滨组相比,肿瘤体积差异无统计学意义(P>0.05)。(2)通过比较各个组荷瘤裸鼠体重,发现与吉西他滨组相比,乌力地格组体重较大,差异有统计学意义(P<0.05);与生理盐水组相比,乌力地格组体重较小,差异有统计学意义(P<0.05);而乌力地格组与DMSO组相比,体重差异无统计学意义(P>0.05)。(3)与生理盐水组及DMSO组相比,乌力地格组的血清CEA、CA19-9含量均较低,差异有统计学意义(P<0.05);乌力地格组与吉西他滨组相比,CEA、CA19-9含量差异无统计学意义(P>0.05)。(4)HE染色验证造模成功后用免疫组织化学染色法检测,发现与生理盐水组、DMSO组相比,乌力地格组PI3K、AKT、mTOR蛋白均表达下降,差异有统计学意义(P<0.05);乌力地格组与吉西他滨组相比,差异无统计学意义(P>0.05)。(5)qRT-PCR检测显示,与生理盐水组和DMSO组相比,乌力地格组中PI3K、AKT、mTOR的RNA相对表达量下降,差异有统计学意义(P<0.05);而乌力地格Objective To explore the effect of Mongolian medicine ulidig on hilar cholangiocarcinoma cell line QBC939 tumor-bearing nude mice and its relationship with PI3K/AKT/mTOR signaling pathway.Methods The nude mice bearing QBC939 tumor were randomly divided into four groups:gemcitabine group,normal saline control group,Dimethy sulfoxide(DMSO)vehicle control group and ulidig group.The drugs were administered based on the group-ings,and the changes in transplanted tumor volume and the weight of tumor-bearing nude mice in each group were doc-umented.After 15 days of continuous treatment,the tumor-bearing nude mice were euthanized following eyeball blood collection.The serum levels of tumor markers CEA and CA19-9 were measured using an enzyme-linked immunosorbent assay(ELISA).Some tumor tissues of tumor-bearing nude mice were stained with HE and immunohistochemistry to observe the staining,structural morphology,necrosis degree of tumor cells,and the expression of PI3K,AKT and mTOR proteins.The relative RNA expression of PI3K,AKT and mTOR in the rest of the tumor tissues was deter-mined by qRT-PCR.Results(1)The volume of transplanted tumors in the ulidig group was smaller than that in both the saline and DMSO groups(P<0.05).However,no significant difference in tumor volume was observed between the ulidig and gemcitabine groups(P>0.05).(2)A comparison of body weights across the groups revealed that the ulidig group had a significantly higher body weight than the gemcitabine group(P<0.05).However,the body weight in the ulidig group was significantly lower than that of the saline group(P<0.05).However,there was no significant differ-ence in body weight between the ulidig group and the DMSO group(P>0.05).(3)However,there was no significant difference in body weight between the ulidig and DMSO groups(P>O.O5).Additionally,serum levels of CEA and CA19-9 in the ulidig group were significantly lower compared to the saline and DMSO groups(P<0.05).No significant difference in CEA and CA19-9 levels was observed between the gemcitabine

关 键 词：肝门部胆管癌 PI3K/AKT/MTOR QBC939 

分 类 号：R29[医药卫生—民族医学]