Sialin(SLC17A5)在腮腺、颌下腺及颌下腺细胞系HSG表达的初步分析

目的研究唾液酸转运蛋白Sialin在正常涎腺组织和颌下腺细胞系HSG的表达.方法应用基因芯片检测正常人腮腺、颌下腺组织Sialin编码基因SLC17A5的表达;提取腮腺、颌下腺组织及颌下腺细胞系(HSG)总RNA,通过RT-PCR在基因水平上验证SLC17A5基因的表达;提取HSG细胞总蛋白,用免疫印迹法检测Sialin在蛋白水平表达.结果基因芯片检测到Sialin编码基因SLC17A5基因在正常人腮腺、颌下腺均有表达,表达比值分别为226、206.9,不存在表达差异;RT-PCR验证芯片结果并在其他涎腺样本及颌下腺细胞系HSG中能扩增出其全部翻译区1485bp的片段,应用商业化的抗体能够在HSG细胞检测到Sialin相应的约54KDa蛋白.结论正常腮腺、颌下腺组织和颌下腺细胞系HSG均表达SLC17A5基因,但是唾液酸转运蛋白Sialin在组织、细胞内定位和功能需要进一步研究.

Expression of Sialin in parotid and submandibular gland of normal human and human submandibular gland cell line(HSG)

Objective To study the expression of an anion transporter sialin in parotid and submandibular glands of normal human and human submandibular gland cell line(HSG).Methods RNA was extracted from parotid and submandibular gland samples of normal human and converted to double stranded cDNA and then to biotinylated cRNA. The biotinylated cRNA was fragmented, and hybridized to Gene Chip (Affymetrix U133 Plus 2.0). RNA was extracted from parotid,submandibular gland samples and HSG cell and RT-PCR was employed to study expression of SLC17A5 .Total protein of HSG cell was obtained and Western blotting employed to study expression of protein of sialin.Results The present value of(SLC17A5)of parotid and submandibular gland of normal human were 226,206.9,respectively.A 1485bp band was amplified from parotid and submandibular gland samples and human submandibular gland cell line(HSG). A 54KDa band was found in total proteins of HSG cell with the antibody of Sialin. Conclusion SLC17A5 is expressed in human parotid and submandibular glands and human submandibular gland cell line(HSG).