Huerta S, Srivatsan E S, Venkatasan N, Livingston E H
VAGLAHS, West Los Angeles/UCLA School of Medicine Division of General Surgery, California 90073, USA.
Dig Dis Sci. 2001 Sep;46(9):1884-91. doi: 10.1023/a:1010626929411.
Expressive loss of the tumor suppressor deleted in colon cancer (DCC) may be superior to lymph node status in predicting patient survival for intermediate stage colon cancer. A polymerase chain reaction (PCR)-based method for detecting DCC would be ideal as a prognostic indicator. DCC is an alternatively spliced molecule; thus, reliability of a PCR test for DCC will depend on amplifying only those regions of the molecule that are lost in the progression of colon cancer. For this reason, we studied a colon cancer cell line model at different stages of tumor progression to determine the alternative splice pattern for DCC. A commercially available colon cancer cell line system at different stages of tumor progression was used to identify which DCC exons are lost by western blot analysis, PCR, and RT-PCR techniques. Colon cancers express abnormal DCC transcripts. The proximal and distal exons are present (exons 2 and 28-29). Exons located in the center of the molecule are absent (6-7 and 18-23). This correlated to DCC protein loss in the cell lines. For clinical utility as a disease marker, exons in the middle portion of the DCC molecule that are spliced out should be utilized. Amplification of the proximal and distal regions will result in falsely concluding that DCC is present when its protein product is not expressed.
在预测中期结肠癌患者生存率方面,结肠癌缺失的肿瘤抑制因子(DCC)表达缺失可能优于淋巴结状态。基于聚合酶链反应(PCR)的检测DCC的方法作为一种预后指标将是理想的。DCC是一种可变剪接分子;因此,DCC的PCR检测的可靠性将取决于仅扩增该分子在结肠癌进展过程中缺失的那些区域。出于这个原因,我们研究了处于肿瘤进展不同阶段的结肠癌细胞系模型,以确定DCC的可变剪接模式。利用一种处于肿瘤进展不同阶段的市售结肠癌细胞系系统,通过蛋白质印迹分析、PCR和逆转录PCR技术来确定哪些DCC外显子缺失。结肠癌表达异常的DCC转录本。近端和远端外显子存在(外显子2和28 - 29)。位于分子中心的外显子缺失(6 - 7和18 - 23)。这与细胞系中DCC蛋白缺失相关。为了作为疾病标志物具有临床实用性,应利用DCC分子中部被剪接掉的外显子。扩增近端和远端区域会导致在DCC蛋白产物未表达时错误地得出DCC存在的结论。