肾透明细胞癌CD133的表达及其体外对α-干扰素和5-氟尿嘧啶处理效果的影响

Expression of CD133 in clear cell renal cell carcinoma cells and the related drug resistances

目的:观察肾透明细胞癌中是否存在类似肿瘤干细胞(TSC)特性的细胞,并探讨这部分细胞对干扰素及化学治疗肾透明细胞癌疗效的影响。方法:选用高转移性肾透明细胞癌细胞株RCC05-TXJ、低转移性肾透明细胞癌细胞株RCC05-ZYJ、临床原位肿瘤样本培养的低代数非转移性肾透明细胞癌细胞(来自男、女性患者各1例)。采用流式细胞术及免疫组化法检测上述4种细胞干细胞标志物CD133的表达情况;采用α-干扰素、5-FU分别处理4种细胞,用MTT法检测药物处理前后细胞存活率。结果:流式细胞术检测结果发现转移性的RCC05-TXJ及RCC05-ZYJ细胞有CD133表达,男、女性原位癌细胞CD133几乎不表达;免疫组化结果显示RCC05-TXJ及RCC05-ZYJ在细胞膜上局部表达CD133,男、女性肾透明细胞癌细胞几乎不表达CD133;化疗药和干扰素干预组RCC05-TXJ、RCC05-ZYJ均出现撤药后24h细胞存活率明显反弹现象,而男、女性原位癌细胞在撤药后细胞存活率持续降低。结论:肾透明细胞癌中存在少数CD133阳性具TSC特性的细胞,这些细胞可能是影响干扰素及化疗效果的原因之一;针对这部分细胞的治疗有望成为今后临床药物治疗的新方向。

Objective： To observe the expression of CD133 in the clear cell renal cell carcinoma （RCC） and to investigate the related drug resistance. Methods： Flow cytometry and immunohistochemical method were used to examine the expression of CD133 in metastatic RCC cell line RCC05-TXJ, low metastatic RCC cell line RCC05-ZYJ and two clinical non-metastatic RCC primary cultures isolated from a male and a female patient. The four ceil lines were treated with IFN-α and 5-FU and the viability of cells were examined by MTT assay before and after treatment. Results： Flow cytometry showed that RCC05-TXJ and RCC05-ZYJ expressed CD133. The in situ carcinomas of male and female patients hardly expressed CD133. lmmunohistochemistry examination showed that the local membrane of RCC05-TXJ and RCC05-ZYJ cells expressed CD133. RCC cells of female and male patients hardly expressed CD133. RCC05-TXJ and RCC05-ZYJ cells exposed to IFN-α or 5-FU showed a rebound of survival 24 h after withdrawal of drugs. The survival rates of RCC cells of male and female patients kept at a low level after withdrawal of drugs. Conclusion： There are a small number of CD133＋ RCC cells in RCC patients, with a property similar to tumor stem cells, which might be one of the important factors influencing the immunotherapy and chemotherapy of tumors.