肿瘤相关肝星状细胞促进肝癌细胞上皮间质转化和迁移

被引:3
作者
孙斌 [1 ]
陈磊 [1 ]
刘振宇 [2 ]
钱海华 [1 ]
施乐华 [2 ]
殷正丰 [1 ]
机构
[1] 第二军医大学东方肝胆外科医院分子肿瘤研究室
[2] 第二军医大学东方肝胆外科医院肝外四科
关键词
肝肿瘤; 肝星状细胞; 上皮间质转化; 细胞增殖; 细胞运动;
D O I
暂无
中图分类号
R735.7 [肝肿瘤];
学科分类号
100112 [医学生物化学与分子生物学];
摘要
目的建立人肝癌组织中肿瘤相关肝星状细胞(tHSCs)分离、培养方法,探讨tHSCs对肝癌细胞生物学行为的影响。方法采用密度梯度离心法从新鲜切除的人肝癌组织样本中分离tHSCs锥虫蓝染色检测细胞活力,细胞免疫荧光鉴定活化tHSCs表达α-平滑肌肌动蛋白的特征,光镜观察培养的tHSCs形态学变化;tHSCs无血清培养后,收集条件培养液,分别与肝癌细胞系QGY-7701、BEL-7402、SMMC-7721共培养,并以肝星状细胞系LX-2做对照,采用MTT法、细胞划痕实验、蛋白质印迹等方法检测肝癌细胞增殖、迁移和上皮间质转化(EMT)相关指标的表达。结果从肝癌组织中分离的tHSCs活率在95%以上,纯度接近100%,在体外可进行传代培养与冻存。tHSCs能诱导3个受试肝癌细胞系产生典型的EMT样形态学变化及其分子标记物表达改变,表现为上皮标记物E-钙黏蛋白表达减少(P<0.05),间质标记物波形蛋白表达增加(P<0.05)。与此同时,tHSCs能促进培养的3个肝癌细胞系增殖和迁移。结论从人肝癌组织中分离的原代tHSCs活力和纯度高,可用于tHSCs的生物学功能实验研究。tHSCs在诱导肝癌细胞产生EMT样改变的同时,体外促进肝癌细胞的增殖和迁移。
引用
收藏
页码:465 / 470
页数:6
相关论文
共 9 条
[1]
活化肝星状细胞与肝细胞肝癌发生及转移的相关性 [J].
王伟 ;
官阳 ;
杨木兰 ;
张春明 .
临床与实验病理学杂志, 2007, (04) :492-494
[2]
The tumor microenvironment in hepatocellular carcinoma (Review).[J].Giulia Leonardi;Saverio Candido;Melchiorre Cervello;Daria Nicolosi;Fabio Raiti;Salvatore Travali;Demetrios Spandidos;Massimo Libra.International Journal of Oncology.2012, 6
[3]
Clinicopathological Significance of ZEB1 Protein in Patients with Hepatocellular Carcinoma [J].
Zhou, Yan-Ming ;
Cao, Lu ;
Li, Bin ;
Zhang, Rui-Xiu ;
Sui, Cheng-Jun ;
Yin, Zheng-Feng ;
Yang, Jia-Mei .
ANNALS OF SURGICAL ONCOLOGY, 2012, 19 (05) :1700-1706
[4]
Icaritin induces cell death in activated hepatic stellate cells through mitochondrial activated apoptosis and ameliorates the development of liver fibrosis in rats [J].
Li, Jing ;
Liu, Peng ;
Zhang, Ruixiu ;
Cao, Lu ;
Qian, Haihua ;
Liao, Jian ;
Xu, Wen ;
Wu, Mengchao ;
Yin, Zhengfeng .
JOURNAL OF ETHNOPHARMACOLOGY, 2011, 137 (01) :714-723
[5]
Gene expression profiles during activation of cultured rat hepatic stellate cells by tumoral hepatocytes and fetal bovine serum [J].
Xia, Yunhong ;
Chen, Rongxin ;
Song, Zhenji ;
Ye, Shenglong ;
Sun, Ruixia ;
Xue, Qiong ;
Zhang, Zhe .
JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY, 2010, 136 (02) :309-321
[6]
Cancer and the tumor microenvironment: a review of an essential relationship [J].
Mbeunkui, Flaubert ;
Johann, Donald J., Jr. .
CANCER CHEMOTHERAPY AND PHARMACOLOGY, 2009, 63 (04) :571-582
[7]
Replicative senescence of activated human hepatic stellate cells is accompanied by a pronounced inflammatory but less fibrogenic phenotype [J].
Schnabl, B ;
Purbeck, CA ;
Choi, YH ;
Hagedorn, CH ;
Brenner, DA .
HEPATOLOGY, 2003, 37 (03) :653-664
[8]
Cytokine receptors and signaling in hepatic stellate cells [J].
Pinzani, M ;
Marra, F .
SEMINARS IN LIVER DISEASE, 2001, 21 (03) :397-416
[9]
Transforming growth factor-β1 regulates platelet-derived growth factor receptor β subunit in human liver fat-storing cells.[J].Massimo Pinzani;Alessandra Gentilini;Alessandra Caligiuri;Raffaella de Franco;Giulia Pellegrini;Stefano Milani;Fabio Marra;Paolo Gentilini.Hepatology.1995, 1