CAJ | 학술논문

目的利用荧光素酶标记的McA-RH7777肝癌细胞建立可供活体观察的大鼠肝癌模型,在此基础上观察选择性结扎门静脉和胆管对肿瘤生长和转移的影响.方法以pCDH-puromycin-CMV为载体将荧光素酶基因转染至大鼠McA-RH7777肝癌细胞,皮下接种Buffalo大鼠,取瘤块异体种植于大鼠肝脏左外叶,活体观察肿瘤生长状态.肝癌建模成功后,随机将大鼠分为三组,即种植叶门静脉联合胆管双结扎组、单纯门静脉结扎组和假手术组.分别于结扎后1周和2周处死,解剖肝肺肾,观察肝内外肿瘤生长和转移情况.结果皮下和肝内种植Buffalo大鼠的成瘤率均为100％.转染荧光素酶Luc-McA-RH7777肝内种植2周后,接种肝叶活体观察可见荧光信号.荧光信号范围和强度随时间的推移增加,4周时肿瘤仍呈局部生长,未见明显肝内和肺转移.门脉联合胆管结扎术后不仅原位肿瘤体积增大,并且于结扎后2周出现肺转移.免疫组化显示肝内瘤灶绝大多数肿瘤细胞呈甲胎蛋白(AFP)免疫反应阳性,部分瘤细胞显示Luc阳性.结论 Luc-McA-RH7777细胞能在Buffalo大鼠肝内成瘤并活体显示,门脉联合胆管结扎可加快肝癌生长和转移.
목적 이용형광소매표기적McA-RH7777간암세포건립가공활체관찰적대서간암모형,재차기출상관찰선택성결찰문정맥화담관대종류생장화전이적영향.방법 이pCDH-puromycin-CMV위재체장형광소매기인전염지대서McA-RH7777간암세포,피하접충Buffalo대서,취류괴이체충식우대서간장좌외협,활체관찰종류생장상태.간암건모성공후,수궤장대서분위삼조,즉충식협문정맥연합담관쌍결찰조、단순문정맥결찰조화가수술조.분별우결찰후1주화2주처사,해부간폐신,관찰간내외종류생장화전이정황.결과 피하화간내충식Buffalo대서적성류솔균위100％.전염형광소매Luc-McA-RH7777간내충식2주후,접충간협활체관찰가견형광신호.형광신호범위화강도수시간적추이증가,4주시종류잉정국부생장,미견명현간내화폐전이.문맥연합담관결찰술후불부원위종류체적증대,병차우결찰후2주출현폐전이.면역조화현시간내류조절대다수종류세포정갑태단백(AFP)면역반응양성,부분류세포현시Luc양성.결론 Luc-McA-RH7777세포능재Buffalo대서간내성류병활체현시,문맥연합담관결찰가가쾌간암생장화전이.
Objective To establish a luciferase labeled McA-RH7777 hepatoma rat model,which could be used for gross observation to further observe the effect of selective ligation of the portal vein and bile duct on tumor growth and metastasis.Methods The luciferase gene was transfected into rat McA-RH7777 hepatoma cells with pCDH-puromycin-CMV as the carrier,which were subcutaneously inoculated into Buffalo rats.Tumor pieces were then heterotransplanted into the left lateral lobe of the allogenic rat liver to observe the tumor growth in vivo.After the successful hepatoma modeling,the rats were randomly divided into three groups,namely the implanted portal vein group with combined portal vein and bile duct ligation,the implanted portal vein group with single portal vein ligation and sham operation group.The rats were executed at the 1 st week and 2nd week after ligation,and the livers were dissected to record the tumor growth and metastasis inside and outside the liver,respectively.Results The tumor formation rates of Buffalo rats after subcutaneous and intrahepatic implantation were both 100％.The fluorescence signal implanted into the liver lobe could be observed in vivo after the intrahepatic implantation of luciferase transfected Luc-McA-RH7777 at 2nd week,the range and intensity of which increased over time.Only local tumor growth could be found at the 4th week,without obvious intrahepatic and lung metastasis.However,both an increased in situ tumor volume and the pulmonary metastasis could be observed in the implanted portal vein group with combined portal vein and bile duct ligation at 2nd week after the ligation.Immunohistochemistry showed AFP positive immunoreactions in the vast majority of intrahepatic tumor cells and Luc positive immunoreactions in part of tumor cells.Conclusion Luc-McA-RH7777 cells could be used to establish the heptoma rat model and the in vivo analysis within the Buffalo rat liver demonstrated that the combined ligation of the portal vein and bile duct can accelerate the development and metastasis of liver cancer.