Helicobacter pylori cagE 유전자 다형성이 위상피세포주에서 Interleukin-8 유도능에 미치는 영향 = Investigation of Helicobacter pylori cagE Locus Diversity on Interleukin-8 Induction in AGS Cell-line

다국어 초록 (Multilingual Abstract)

Helicobacter pylori infection is highly prevalent, as high as 2/3 of whole population infected, in Korea. H. pylori infection initiates inflammation by induction of interleukin-8 through type IV secretion of CagA. It was recently suggested that induction failure of IL-8 is not associated with defect in cag PAI but associated with cagE locus diversity. This study was designed to investigate ability of 11-8 in-duction according to sequence variation within the cagE gene, cagA TP motifs and vacA m-types in vitro study using AGS cell-line, and to evaluate its association with different clinical outcome. Seventy-four H. pylori stains were isolated from 23 patients with gastric cancer (Ca), 24 subjects with gastritis (G) and 27 patients with duodenal ulcer (Du) in Kangnam St. Mary's Hospital, Seoul, Korea. cagE gene diversity was confirmed by the PCR-RFLP methods with MboI/NlaIII and tyrosine phosphate motifs (TPMs) of cagA was determined TPM-A and C by using DdeI/Tsp5091 restriction enzyme and TPM-B was determend by Real time PCR the method of Owen et al. and IL-8 was measured by ELISA assay. IL-8 activity was positively detected in 59 among 74 strains $(79.7\%)$. IL-8 secretion was significantly increased in MboI A and MboI B type compared to MboI C type and in MboI/NlaIII A-C and B-C type than C-C type. 1L-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. There was no significant difference in IL-8 activity among patient groups. cagE gene diversity is thought to be mainly associated with the induction of IL-8 in H. pylori infection.

국문 초록 (Abstract)

한국에서 분리한 Helicobacter pylori 균주가 위암 세포주에서 IL-8를 유도하는 유도능을 관찰하고자 하였으며, 특히 4형 분비구조의 주요 구성체인 cagE 유전자의 다형성에 따라 IL-8 유도능의 차이 및 각 질환에서 분리한 균주에서의 차이가 있는지 검토하고자 하였다. 강남성모병원 내시경검사를 받은 환자들 중 H. pylori 감염 양성으로 확인된 위암 23명 $(31.0\%)$, 위염 24명$(32.6\%)$, 십이지장궤양27명$(36.4\%)$ 총 74명을 대상으로 하여 H. pylori를 분리한 후 PCR-RFLP방법을 이용하여 cagE 다형성을 확인하였으며, PCR 및 Real-time PCR을 이용하여 ragA TPMs를 결정하였다, 또한 분리된 균주를 위암세포주에 감염시켜 상등액을 취하여 ELISA방법으로 IL-8을 정량화 하였다. 전체 74개의 분리 균주 중 59개의 균주$(79.7\%)$에서 IL-8 유도능이 있는 것으로 확인되었으며, cagE 다형성 중에서 가장 높은 빈도로 관찰되는 형태는 MboI/NlaIII B-C, A-C 및 C-C 타입 이었으며, 이중 A-C와 B-C 형태가 C-C 형태에 비해 유의적으로 높은 IL-8 유도능을 나타냈다. 그러나 cagA TPMs의 분류 및 vacA m-아형 분류와 IL-8 유도능과는 연관성이 없었다. 비록 본 연구에서 cagE 다형성과 IL-8 유도능과 아주 밀접한 연관성이 나타나지는 않았지만 vacA m-아형이나 cagA TPMs 보다는 cagE 다형성이 IL-8 유도능과 연관성이 클 것으로 생각된다.

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