摘要
目的建立裸小鼠人肝癌原位移植多药耐药模型，探讨其生物学特征及耐药机制。方法裸小鼠人肝癌（Bel-7402）原位移植，用阿霉素腹腔注射诱导耐药，观察其病理形态学特征，采用荧光定量RT．PCR检测多药耐药基因MDR1mRNA在肝癌组织中表达。以流式细胞仪检测癌细胞MDR1基因产物P-糖蛋白（Pgp）的表达，并通过免疫组化和激光共聚焦显微成像系统对Pgp进行细胞定位及形态观察。结果移植瘤组织形态及生物学行为符合人肝癌特征，实验组MDR1mRNA表达量在1．4×10^3～3．6×10^6copy／μgRNA，阳性率达70．00％，PgP表达为75．45％±5．67％；而对照组MDR1mRNA表达阴性，Pgp表达阳性率为4．25％±1．28％（P〈0．01）。形态学观察显示Pgp大多位于细胞膜。结论裸小鼠人肝癌原位移植模型在形态及生物学特征上能高度模拟人肝癌。阿霉素在体内主要通过诱导人肝癌细胞多药耐药MDR1mRNA基因及影响功能蛋白Pgp的表达。获得耐药性。
Objective To study the pathomorphological characteristics and muhidrug resistance（MDR） mechanism of MDR model of human hepatocellular carcinoma （ HCC ） orthotopic transplanted into nude mice. Methods foursix week nude mice was orthotopic transplanted with HCC tissue （Bel-7402） and treated by injection of adrimycin （ ADM 1.5 mg/kg, for 8 weeks） into the abdomen. The expression of MDR1 mRNA of nude mice liver tumor was detected by fluoregenic probe quantitative RT-PCR. The p-glycoportein（Pgp） expression was examined by flow cytometric assay and S-P immunohistochemical methods. Laser scanning confocal microscopy was used to check Pgp orientation in HCC cell. Results The pathomorphological results showed that the most typical behaves of HCC were observed in nude mice liver tumor. The MDR1 mRNA level of ADM group was 1.4 × 10^3 -3.6 ×10^6 copy/μg RNA with 70. 00% positive expression, but in control group was negative. The pgp level of ADM group was 75.45% ±5.67%, but that in control group was 4. 25% ± 1.28% （P 〈0. 01 ）. Pgp orientation was on the luminal side of their surface membrane in HCC cell. Conclusions The typical morphological characters of HCC were observed in the multidrug resistance model of orthotopic transplantation of HCC. The main mechanism of multidrug resistance is correlated with the expression degree of MDR1 mRNA and Pgp in HCC cell.
出处
《安徽医科大学学报》
CAS
北大核心
2007年第1期5-8,共4页
Acta Universitatis Medicinalis Anhui
关键词
肝肿瘤/病理学
疾病模型
动物
小鼠
裸
抗药性
多药
基因
MDR
肿瘤移植
liver neoplasms/pathology
disease model, animal
mice, nude
drug resistance, multiple
genes, MDR
neoplasm transplantation
