摘要
目的寻找鼻咽癌中差异性表达基因,包括与鼻咽癌发病相关的候选抑瘤基因。方法应用cDNA代表性差异分析法(RDA),分离原代培养的正常人鼻咽上皮细胞与鼻咽癌细胞株HNE1中差异表达的cDNA序列,Southern杂交和Northern杂交被用来分析差异性表达产物的来源,最后,将这些序列克隆到pGEM-Teasy载体中,并用链终止法测序。结果在第4轮杂交及扩增反应后,获得4条差异性条带。Southern杂交及Northern杂交证明,这些差异性片段来自作为“检测”扩增子的正常人鼻咽上皮,在鼻咽癌细胞株HNE1中不表达或表达降低。序列分析这些差异性片段的克隆,发现一些序列是与已知基因高度同源的基因,包括一些看家基因;另有一些基因则为新基因序列。结论鼻咽癌的发生是一个多基因参与的过程,所获得的差异性片段中,与之同源的一些已知基因具有抑瘤功能。
Objective To search differentially expressed sequences correlated with pathogenesis of human nasopharyngeal carcinoma(NPC), including the candidates of tumor suppressor genes. Methods cDNA representational difference analysis (RDA) was performed to isolate differentially expressed sequences between cDNA from normal human primary cultures of nasopharyngeal epithelial cells and cDNA from NPC cell line HNE 1. The sources of differentially expressed products were proved by Southern blot and Northern blot. The fragments were cloned with pGEM T easy kit and sequenced by the chain termination reaction.Results Four differentially expressed cDNA fragments were isolated in the fourth subtractive hybridization using cDNA from normal human primary cultures of nasopharyngeal epithelial cells as tester amplicon and cDNA from NPC cell line HNE 1 as driver amplicon by cDNA RDA. These differential cDNA fragments revealed that they really came from the tester amplicon and were not expressed or down regulated in the NPC HNE 1 cells. Of these obtained clones, some are the fragments of the human known genes including house keeping genes, the others are novel genes.Conclusion NPC involves alteration of multiple genes. Some of known genes matched with the differentially expressed sequences have an effective suppressive ability on the carcinoma.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
1998年第6期341-344,共4页
Chinese Journal of Medical Genetics
基金
国家863高技术计划
湖南省自然科学基金
医学遗传学国家重点实验室开放经费资助
关键词
鼻咽癌
抑瘤基因
CDNA
差异分析法
Nasopharyngeal carcinoma
Tumor suppressive gene
cDNA representational difference analysis