大肠癌与正常大肠粘膜中新分化相关基因地表达.docxVIP

在大肠癌中存在着普遍的不同程度的表达缺陷，并且与大肠癌分化程度呈正相关，与淋巴结转移关系密切，可能是人大肠癌分化的 一个新的分子标记。继续深入探讨此大肠癌分化相关基因及其产 物与大肠癌及其它肿瘤发生发展的关系，将具有临床实际意义。 关键词：大肠肿瘤；细胞分化；基因表达；聚合酶链反应 ·5‘ Differential Expression of a Novel Colorectal Cancer Differentiation—related Gene in Colorectai Cancer and Autologous Healthy Mucosa Butyrate iS an important colonic fuel and induces differenti— ation in colonic eell lines．cDNA for a novel colorectal cancer (CRC)differentiation—related gene，designated SBA2(Gen— Bank accession No：AF229181)，has been identified in human CRC cell line CloneA after modulation by sodium butyrate．It consisted of 2470 nucleotides and an open reading frame(0RF) encoded 404 amino acid residues with fl calculated molecular weight of 44．4 k Da．The deduced amino acid sequences showed significant homology tO mouse SWiP一2(96％)，mouse WSB一2 (95％)and human WS1(52％)，which belong tO a new family of the suppressor of cytokine signaling(SOCS)．The SBA2 pro— tein product may be a new member of SOCS protein family nega— tively regulating cytokine signal transduction．We investigated the use of reverse transcription—polymerase chain reaction (RT—PCR)for the relative quantification of SBA2 expression jn 4 human CRC cell lines with different grades of differentiation and in 30 surgical specimens of CRC and autologous healthy mu— cosa． We have developed an efficient protocol for relative quan— tification of gene expression in both clinical samples and cell lines of CRC by semi-quantitative RT—PCR．Linear evaluation ·6· of gene expression over a wide range was achieved by plotting the amplification curves of the sequence of interest and the p actin sequence that served as an internal contr01．Determination of the optimal parameters of an exponential range was carried out by termination the PCR at sequential cycles and by serially diluting the amount of eDNA． The yield of PCR pl+oduct is proportional to the starting amount of the template only under conditions in which PCR am— plifieation proceeds e