| 大鼠肝癌细胞表面特标记与成瘤能力有关
| | 医学社会实践论文【关键词】 ,肝肿瘤
Relationship between surface markers characteristic and tumorigenic ability in rat hepatocarcinoma cells
【Abstract】 AIM: To study the relationship between surface markers characteristic and tumorigenic ability in rat hepatocarcinoma cells. METHODS: The primary liver carcinoma in rats were induced by diethylnitrosamine. Tumor cells from 8 rats with primary liver carcinoma were separated according to rat oval cell (OVC) surface markers such as CD34, cKit, Thy1, AFP, CK7, CK8,CK14,CK18,CK19 and GGT.The tumor in nude rats which had been injected with hepatocarcinoma cells (10 OVC marker negative or positive cells ) for 1 month were weighted respectively. The subpopulations of hepatocarcinoma cells, which had a stronger ability to cause tumor after injected to nude rats, were diluted to 1/100 or 1/10 of original density. Then we analyzed the tumorigenic ability of these subpopulations in a low cell density. RESULTS: The weight of the tumors in nude rats which had been injected with the CD34, Thy1, AFP, CK7, CK8 and GGT positive subpopulations was different from that of ones with the negative subpopulations (P<0.05 or 0.01). The cell subpopulation of AFP(+) or CK7(-) had a stronger ability to cause tumor when diluted to 1/100-1/10 of original density. CONCLUSION: Different subpopulations of the hepatocarcinoma cells had a different ability to cause tumor in rats. AFP(+) and CK7 (-) may be the markers characteristic of hepatocarcinoma stem cells.
【Keywords】 liver neoplasms; stem cells; surface markers
【摘要】 目的医学社会实践论文：研究大鼠肝癌细胞表面标记特征与成瘤能力关系. 方法：二乙基亚硝胺为诱癌剂制造大鼠肝癌模型，分离培养肿瘤细胞，按照卵圆细胞表面标记（CD34,cKit,Thy1,AFP，CK7，CK8,CK14,CK18,CK19和GGT）分选肿瘤细胞，比较各个表面标记阳性肿瘤细胞亚群与阴性肿瘤细胞亚群移植裸鼠后成瘤能力差异，并且对数稀释成瘤能力强的医学社会实践论文细胞亚群，分析低密度下强成瘤能力细胞亚群与相应阴性细胞亚群成瘤差异. 结果： CD34,Thy1,CK7,CK8,AFP和GGT等6个标记的阳性与阴性肿瘤细胞移植裸鼠后，成瘤能力差异显著(P<0.05或0.01)；1/100~1/10细胞密度的CK7阴性与AFP阳性细胞亚群成瘤能力仍然比其对应细胞亚群为高. 结论：大鼠原发性肝癌肿瘤细胞中存在成瘤能力差异巨大的肿瘤细胞亚群. CK7(-)和AFP(+)，可能是大鼠肝癌干细胞的部分表面标记特征.
【关键词】 肝肿瘤；干细胞；表面标记
0引言
肿瘤干细胞(tumor stem cells, TSC)是近年肿瘤科学研究的热点. TSC来源于相应的成体干细胞，是肿瘤发生、扩散与转移的基础［1-3］. 目前肝癌干细胞的研究报道很少. 我们知道，卵圆细胞（oval cells, OVC）不仅具备基本的肝干细胞性质，而且与肝癌发生关系密切，是最接近肝癌干细胞的成体干细胞. 我们通过OVC表面标记分选肝癌肿瘤细胞，了解不同肝癌细胞亚群的裸鼠成瘤能力，探索肝癌干细胞的表面标记特征.
1材料和方法
1.1材料雄性清洁级SD大鼠8只， 8 wk大，体质量（233.8±24.5）g，雄性裸小鼠(BALB/cnu/nu)，4~6 wk，体质量15~20 g，由南方医科大学动物实验中心提供. 饲养温度22~25℃，湿度（50±5）%，在正常光照及自由摄食条件下饲养于普通饲养室，裸小鼠饲养于SPF级饲养室. RPMI 1640与胎牛血清为美国Gibco公司产品; 二乙基亚硝胺及胰蛋白酶为Sigma公司产品;一抗CD34, cKit,Thy1,AFP,CK7,CK8,CK14,CK18,CK19和GGT购自北京中杉金桥生物技术有限公司.
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