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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。目前仍需继续深入研究该两基因的功能
Objective] To find new factors related to the occurrence and development of colorectal cancer at the molecular level and to study the structure and function of two negative colorectal cancer related genes. [Methods] Two new colorectal cancer gene loci were mapped by enhanced fluorescence in situ hybridization (FISH). The new gene function was studied by Dot blot, Northern blot, RT-PCR_ELISA, immunohistochemistry and transfection experiments in vitro and in vivo. [Results] The full-length cDNA sequence analysis and chromosomal localization studies of the two new genes have been completed. The nucleic acid and protein expression and functional studies of the two genes have been performed. The SNC6 gene has a full length of 3168bp and is located in the chromosome 22q13 band with a total of 12 exons and 11 introns. SNC19 has a full length of 3152bp and is located on chromosome 11q24. The two genes were named ST13 and ST14 genes by the International Nomenclature Commission in 1998 and included in the human genome map. We compared the expression of two novel genes in normal colorectal mucosa and cancerous tissue, and the cancer tissue was lower than normal mucosa and expressed differently in different tumor cell lines and tissues. Both genes have obtained the expressed protein. Monoclonal antibody has been prepared in SNC6. The expression of nucleic acid is similar to that of nucleic acid in immunohistochemistry. SNC6 gene has been shown to inhibit the growth of colorectal cancer cell lines in vitro and in vivo. SNC19 has been identified as a member of the serine protease family. [Conclusion] The SNC6 and SNC19 genes can be used as new colorectal cancer related genes for research and utilization. It is still necessary to further study the function of the two genes.