目的探讨人肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)是否存在与细胞生长调节相关的转录因子19(TCF19)选择性剪接异构体,探索其表达特点及其与ccRCC发生的关系。方法通过选择性剪接数据库(alternative Spli-cing Database,ASD)对TCF19基因可能存在的选择性剪接异构体种类进行预测。设计可能存在的5种异构体引物,采用RT-PCR技术,在32例原发肾透明细胞癌的混合样本及癌旁组织的混合样本中分别检测预测异构体的表达,并对电泳所获条带进行克隆、测序。应用RT-PCR定量检测在各个组织样本中异构体TCF19-1、TCF19-3在原发肾癌及癌旁组织的表达差异。结果 ccRCC组织中存在2种新型TCF19替换剪接异构体(TCF19-1、TCF19-3)。RT-PCR检测结果发现这2种异构体在癌和癌旁组织中均表达,但TCF19-1在原发癌组织中表达率高于癌旁组织(P〈0.001);TCF19-3在癌旁组织中的表达率与原发癌组织比较,差异无统计学意义(P=0.082)。结论首次发现ccRCC组织中存在2种TCF19选择性剪接异构体TCF19-1和TCF19-3,其中TCF19-1的表达可能与ccRCC有关。
Objective To study whether there is expression of the alternative splice variants of TCF19,a cell growth regulatory gene,in human clear cell renal cell carcinoma(ccRCC)tissues and their relationship with the development of ccRCC.Methods Alternative Splicing Database was used to predict the possible alternative splice variants of TCF19.Specific primers for amplifying the 5 predicted variants of TCF19 were designed and used to examine the corresponding variants in 32 ccRCC tissues and normal tissues adjacent to the tumors by RT-PCR technique.The amplicons were then cloned and sequenced.Quantitative RT-PCR was used to detect the expression differences of the variants between the primary renal cell carcinoma and adjacent tissues.Results Two new isoforms of TCF19(TCF19-1 and TCF19-3)were identified in the ccRCC and the adjacent tissues.RT-PCR results showed that TCF19-1 expression was significantly higher in the primary ccRCC tissues than that in the adjacent normal tissues(P0.001).And the expression of TCF19-3 in the primary tumor was only slightly higher than that in the adjacent tissues(P=0.082).Conclusion This study for the first time identifies the existence of two alternative splice isoforms of TCF-19,TCF19-1 and TCF19-3,in human ccRCC tissues,and the expression of TCF19-1 might be related to the development of ccRCC.