肾癌干细胞的表面分子标志及生物学特性研究

More and more recent studies showed that the cancer stem cells(CSCs) play an important role in the growth and propagation of the tumor. CSCs have the abilities of tumorigenic, self-renewal ,chemo- and radioresistant. By using CD44 and CD133 cell markers, CSCs have been successfully isolated from brain, breast, colon, liver, lung, pancreas, ovary and prostate cancer. The complete eradication of CSCs will bring new hope for cure the malignant tumors. Renal carcinoma is one fo the most popular urinary tumors and not sensitive for chemo- and radio theraphy. New theraphys (such as eradication of CSCs )for treat the renal carcinoma are needed to explore. Untill now, no stdudies have found the special molecular markers of the renal carcinoma. Our study will search the special molecular markers of the renal carcinoma and their biological charactars. We have isolated one cell subsets which have self-renewal abilities from renal carcinoma cells by transfection with a plasmid containing the human Oct-4 promoter driving a GFP reporter. Based on the preview studies,we will further isolate more subsets by using other molecular markers, flow cytometry and other technology, and study the abilities of tumorigenic, self-renewal and other biological charactors. Our study will provide the theoretical basis for renal cancer stem cell theraphy.

越来越多证据研究表明肿瘤干细胞（cancer stem cell，CSC）是恶性肿瘤复发、转移的根源，CSC具有高致瘤性及自我更新潜能，且对放化疗不敏感。利用CD44，CD133等细胞表面抗原已成功从脑、乳腺、结肠、肝、肺、胰腺、卵巢、前列腺等多种恶性肿瘤中分离CSC。彻底根除CSC为治愈癌症的带来新希望，肾癌最常见的泌尿系肿瘤之一，对放化疗均不敏感，亟待寻找新的有效治疗途径如杀灭CSC。目前为止尚未发现肾癌CSC可靠而特异的表面标记物。本课题旨在寻找肾癌CSC固有标记分子，并研究它们的生物功能特性。我们运用干细胞因子OCT4全长启动子驱动的GFP表达标记肾癌细胞，已分离出具自我更新的细胞亚群，在此实验基础上，再结合流式细胞仪等其它分子生物学技术及表面标志，进一步分离更多细胞亚群进行自我更新，致瘤能力等生物学测定，此研究将为开发肾癌干细胞治疗策略提供理论依据及操作平台。

本课题完成了肾癌细胞株和病人来源的原代肾癌细胞的获取及培养，并对其进行了SPOP免疫组化检测及VHL基因突变分析，从而发现SPOP和VHL基因突变可能是肾透明细胞癌的分子标记，对其监测将有可能协助肾透明细胞癌的诊断。本课题实验性地运用干细胞因子OCT4全长启动子驱动的GFP表达标记肾癌细胞，并对GFP阳性细胞进行分离、纯化，以明确以OCT4-GFP作为标记的肾癌细胞的干细胞生物特性，进一步寻找新的肾癌干细胞的标志。