摘要
目的:检测WWOX(WW domain containing oxidoreductase)基因在肺腺癌细胞株A549中的变化及其意义.方法:用逆转录-聚合酶链反应(RT-PCR)技术、聚合酶链反应结合二核苷酸重复序列多态性方法、免疫印迹分析(Western blot)方法分别对WWOX基因6～8外显子在A549细胞转录本丢失、杂和性丢失(LOH)和WWOX蛋白表达异常进行检测.结果:A549细胞样本为WWOX基因6～8外显子转录本的丢失,而在对照的原代培养人气道上皮细胞样本(简称原代培养)未发现WWOX基因的6～8外显子的丢失(P<0.05);A549细胞样本中存在D16S3029及D16S3096两个微卫星位点的LOH 而对照的原代培养中无WWOX基因的杂合性丢失(P<0.05);Western blot结果显示WWOX基因在A549细胞的蛋白表达(A值为817464.90±64916.64)明显低于在原代培养中的蛋白表达(A值为2150142.00±64916.64)(P<0.01).结论:WWOX基因可能在肺腺癌的发生发展中起重要作用;转录本丢失、LOH及蛋白低表达均可能是WWOX基因在肺腺癌中的失活方式.
Purpose:To detect the abnormalities of WWOX(WW domain containing oxidoreductase) gene in human lung adenocarcinoma cell line A549. Methods:Deletion of WWOX exons 6-8 transcript was analyzed by reverse transcriptase-PCR technology; loss of heterozygosity (LOH) of WWOX gene was analyzed by PCR-based assays for dinucleotide repeat polymorphisms technology. Aberrant expression of WWOX protein was analyzed by western blot. Results:A549 cells samples showed loss of WWOX exons 6-8 transcript.This deletion was not detected in normal primary cultured human bronchial epithelial cells samples.Three microsatellites(D16S3029、D16S3096、D16S504)did not have LOH in the normal primary cultured human bronchial epithelial cells samples, but D16S2029 and D16S3096 were all found to have LOH in A549 Cells samples. We further observed that expression of WWOX protein was significantly lower in A549 cell samples compared to the normal primary cultured human bronchial epithelial cells samples. Conclusions:WWOX gene may be important during tumorigenesis in lung adenocarcinoma cancer.Deletion of exons 6-8,LOH and aberrant expression of protein are all modes of WWOX gene inactivity in lung adenocarcinoma cancer.
出处
《中国癌症杂志》
CAS
CSCD
2005年第3期234-237,共4页
China Oncology
