摘要
目的]在分子水平寻找与人大肠癌的发生发展相关的新的因素 ,研究两个大肠癌负相关基因的结构与功能。[方法]采用强化荧光原位杂交 (FISH) ,进行两个新大肠癌基因染色体定位。通过Dotblot、Northernblot、RT_PCR_ELISA,免疫组化及体内外转染实验进行新基因功能研究。[结果]已完成两个新基因的全长cDNA序列分析及染色体定位等结构研究 ,并进行了两基因的核酸与蛋白表达研究及功能研究。SNC6基因全长3168bp ,定位于染色体22q13区带 ,共有12个外显子及11个内含子。SNC19全长3152bp,定位于染色体11q24区带。两个基因于1998年被国际命名委员会分别命名为ST13、ST14基因 ,并被列入人类基因组图谱中。对比两个新基因在正常大肠粘膜及癌组织核酸水平表达情况 ,癌组织低于正常粘膜 ,在不同肿瘤细胞系及组织中表达不一。两基因均已获得了表达蛋白。SNC6已制备了单抗 ,经免疫组化检测表达与核酸表达结果相似 ,经体内外研究SNC6基因有抑制大肠癌细胞系生长的作用。SNC19已明确为丝氨酸蛋白酶家族成员。[结论]SNC6与SNC19基因可作为新的大肠癌相关基因加以研究、利用。
To search previously uncertained factors that correlated with the occurrence and development of human colorectal cancer.To study the structure and function of the two colorectal cancer negatively related genes.Enhanced fluorescence in situ hybridization technique was used in chromosome localization.The function of novel genes were studied by Dot blot,Northern blot,RT_PCR_ELISA,in situ hybridization,immunohistochemistry and in vivo and in vitro transfection trial.The structure and function studies of SNC6 and SNC19 genes were carried out.The full length cDNA of SNC6 and SNC19 was 3168bp and 3152bp respectively.The chromosomal localization of SNC6 and SNC19 was in 22q13 and 11q24 respectively.There was 12 exons and 11 introns in the SNC6 gene.The SNC6 and SNC19 genes were respectively nominated as ST13 and ST14 by HUGO Nomenclature Committee,and were assigned to Human Genome Maps.The expression level of SNC6 was lower in colorectal cancer than that in normal intestinal mucosa,and was varied in different cancer cell lines and tissues.The mono_antibody of SNC6 was obtained.The similar results of the expression of SNC6 gene were detected by protein and nucleotide level.It is identified that SNC6 gene could inhibit growth of colorectal cancer cell in vivo and in vitro.It is elucidated that SNC19 encodes a novel human serine protease.[Conclusion] SNC6 and SNC19 gene can be further investigated as a novel colorectal negatively related gene.
出处
《中国肿瘤》
CAS
2001年第3期136-138,共3页
China Cancer
基金
"九五"国家科技攻关项目! (96-906-01-21)
