B超弹性成像监测下应用吉西他滨耐药乳腺癌细胞4T1构建裸鼠乳腺癌肝转移模型  

Application of a gemcitabine-resistant variant of breast cancer cell line(4T1/Gem) to construct nude mouse models of breast cancer with hepatic metastasis under ultrasonic elastography

作　　者：叶志东[1,2] 黄迪[1,2] 黄宇[1] 张强[1] 麦振豪 赵俐[1,2] 古维立[1,2] 

机构地区：[1]广州医科大学附属广州市第一人民医院普通外科,广州510180 [2]广州消化疾病中心,广州510180

出　　处：《广州医药》2016年第1期49-53,共5页Guangzhou Medical Journal

基　　金：广东省科技计划项目(2011B061300024;2013B021800057);广东省自然基金项目(10151006001000013);广州市医药卫生科技一般引导项目(20131A011029;20141A01003);广州市科技和信息化局科普专项项目(2014KP000086)

摘　　要：目的构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型。方法采用低浓度加量持续诱导法,诱导吉西他滨耐药乳腺癌细胞4T1耐药株,命名为4T1/Gem;CCK-8法测定4T1与4T1/Gem细胞的增殖抑制率,计算耐药指数;Western blot法检测细胞P-gp蛋白表达;B超引导下注射4T1/Gem细胞悬液诱导裸鼠肝脏成瘤;HE染色观察肿瘤组织病理情况,免疫组化法检测瘤组织ER、PR、HER2、Ki-67和P-gp蛋白的表达。结果经过14个月的诱导成功建立4T1/Gem细胞株,可在含40μg/m L的Gem培养液中稳定生长。4T1/Gem细胞耐药指数为4T1细胞的788.547倍。与亲代相比,4T1/Gem处于G1期和G2期的细胞增加,S期细胞减少;上调P-gp蛋白的表达。4T1/Gem细胞成功建立裸鼠乳腺癌肝转移模型,瘤组织中ER、PR、HER2蛋白阴性表达,Ki-67阳性10%和P-gp蛋白阳性表达。结论成功构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型,为开发治疗乳腺癌肝转移化疗耐药的药物提供实验基础。Objective To construct a gemcitabine-resistant variant of the breast cancer cell line （4T1/Gem） and establish a nude mouse model of breast cancer with hepatic metastatic. Methods A gemcitabine-resistant variant of the breast cancer 4T1 cell line was induced by gradually increasing the concentration of gemcitabine ; this variant is referred to in this study as 4T1/Gem. The proliferation suppression rates of 4T1 and 4T1/Gem cells were determined by using the CCK-8 essay to evaluate the drug resistance indices of the cell lines. Western blot analysis was used to detect P-gp protein expression. Under ultra-sonography, a 4T1/Gem cell suspension was injected into nude mice to induce liver tumors. H＆E staining was used to observe tumor pathology, and immunohistochemistry was used to detect the expression of ER, PR, HER-2, Ki-67, and P-gp. Results After 14 months of induction, a 4T1/Gem cell line is established successfully. The cell line can grow stably in culture liquid containing 40 ixg/ml gemcitabine. The drug resistance index of 4T1/Gem is 788. 547. Compared with the 4T1 cell line, the 4T1/ Gem cell line can upregulate P-gp protein expression and successfully establish a nude mouse model of breast cancer with hepatic metastatic. ER, PR, and HER-2 proteins exhibit negative expression in the tumor tissue. The positive expression of P-gp and 10% of Ki-67 proteins is also observed. Conclusion This study successfully constructs a gemcitabine-resistant variant of the breast cancer cell line （4T1/Gem） and establishes a nude mouse model of breast cancer with hepatic metastatic, thereby providing an experimental basis for developing and treating a drug-resistant variant of breast cancer.

关 键 词：乳腺癌 吉西他滨 耐药 P—gp蛋白 裸鼠成瘤 

分 类 号：R737.9[医药卫生—肿瘤]