牙龈卟啉单胞菌侵入口腔上皮细胞后基因表达变化的初步研究  

Identification of specific pathogenic genes of Porphyromonas gingivalis during the invasion of oral epithelial cells

作　　者：成伟[1] 王洁[1] 孔令雪[1] 齐霞[1] 吴亚菲[2] 赵蕾[2] 

机构地区：[1]四川大学口腔疾病研究国家重点实验室,四川成都610041 [2]四川大学华西口腔医院牙周科,四川成都610041

出　　处：《牙体牙髓牙周病学杂志》2012年第11期641-645,共5页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金(30801295);教育部博士点专项科研基金青年新教师基金(200806101107)

摘　　要：目的:采用差异显示反转录PCR技术,比较牙龈卟啉单胞菌侵入口腔上皮细胞后基因表达的变化,初步筛选与细菌侵入宿主细胞相关的毒力致病基因。方法:提取纯培养状态和侵入KB细胞后的牙龈卟啉单胞菌ATCC 33277细菌总RNA;反转录PCR技术显示cDNA表达条带;筛选并回收差异表达条带,二次扩增cDNA,经测序和BLAST同源性比对,初步分析细菌侵入KB细胞后表达差异基因的功能。结果:共筛选获得3条牙龈卟啉单胞菌侵入KB细胞后表达存在差异的基因HX 01、HX 02、HX 03;BLAST同源性比对结果显示,HX 01与牙龈卟啉单胞菌ABC转运蛋白编码基因PG 0683具有96%的同源性;HX 02与牙龈卟啉单胞菌PepO编码基因PG 0159具有97%的同源性;HX 03未找到类似同源性基因序列。结论:牙龈卟啉单胞菌在侵入KB细胞后发生了细菌毒力基因的表达变化,跨膜蛋白ABC转座子和PepO可能参与细菌侵入宿主细胞的致病过程。AIM： To identify the specific pathogenic genes of P. gingivalis during the invasion of oral epi- thelial ceils by differential display reverse transcription PCR technology. METHODS： Total RNAs of P. gingivalis ATCC 33277 from control and infected KB cells were isolated, cDNA was synthesized and differential display reverse transcription PCR technology was performed to display the differential expression of cDNA between the two groups. The specific cDNA electrophoretic bands expressed by P. gingivalis from infected cells were harvested and amplified subse- quently. The gene sequences were analyzed and identified based on the gene information from Nucleotide BLAST. RESULTS：Three potential specific pathogenic genes were identified, and were named as HX 01, HX 02 and HX 03. The homologous searching results showed that the homology between HX 01 and P. gingivalis ABC transporter gene en- coding PG 0683, HX 02 and P. gingivalis PepO gene encoding PG 0159 were 96% and 97% respectively, while none of the known gene was matched with HX03. CONCLUSION： Invasion into KB cells could regulate the gene expres- sion of P. gingivalis. Three genes differentially regulated by P. gingivalis were associated with ABC Transposons, PepO transmembrane protein and protein of unknown function, which suggested that ABC Transposons and PepO transmem- brane protein might play a role during the invasion of host cells by P. gingivalis.

关 键 词：牙龈卟啉单胞菌 口腔上皮细胞 侵入 差异显示PCR 

分 类 号：R780.2[医药卫生—口腔医学]