The study showed that Dclk1-expressing cells increased in number in assoication with parietal cell loss. Given the multiple splice variants of Dclk1, the expression of Dclk1 isoforms during gastric carcinogenesis have been examined. It was found that the expression of Dclk1 isoforms are altered in gastric carcinogenesis. SPEM was induced by DMP-777 and the expression of Dclk1 isoforms were examined in translational and transciprtional elvel. While the expression intensity of both LF and SF increased in metaplastic gastric muscosa, the expression of mRNA was dynamic defined to regional location. One intersting finding was that each isofrom was labeled differently even within the same cell. Additionally, in situ hybridization analysis confirmed the altered expression of Dclk1 isoforms in metaplsia by oxyntic atrophy. These findings imply possibilities that Dclk1 isoforms play an important role in gastric carcinogenesis. Furthermore, it was seen that from endoscopic resection specimens of gastric cancer patients, LF was lost and only SF was expressed in cancer. This finding suggest Dclk1 LF as a hall marker in determining the progression of gastric cacner. Altogether, these data provide new cellular and molecular aspects of Dclk1 isoforms in association with different stages of gastric cancer.
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< ABSTRACT (IN KOREAN)>
위암 발생과정에서
Dclk1 isoform 발현의 역동적 변형에 대한 연구
<지도교수 남 기 택>
연세대학교 대학원 의과학과
황 혜 경
Dclk1은 장에서 quiescent stem cell marker로 알려져 있을 뿐만 아니라, 위의 tuft cell의 마커로 알려져 있다. Dclk1은 long form(LF, full length), short form(SF, kinase domain only), 363 isoform (doublecortin domain only)의 3개의 전사체를 가진다. 선행연구에서 cancer와 Dclk1 발현간의 상관관계가 알려져 왔으나, metaplasia와 위암에서 Dclk1 isoform 발현에 대해서 는 보고된 바가 없다. 이에 Dclk1의 isoform 발현이 위암 발생 과정에 관여할 것이라 가 정하에 DMP-777을 투여한 마우스 SPEM 모델을 이용하여 Dclk1의 isoform 발현을 단백 질과 RNA level에서 분석하였다. LF과 SF의 단백질 발현은 위의 전체 부위에서 증가하는 양상을 보였으나, 이에 반하여 RNA의 발현은 부위별로 다양한 발현양상의 변화를 보였 다. 또 Dclk1 발현세포를 immunohistochemistry와 in situ hybridization 방법으로 관찰한 결과, Dclk1 isoform마다 세포 내에 발현하는 위치가 다를 뿐만 아니라 위암 발병 과정 중 Dclk1 isoform에 변화가 보이는 것을 확인하였다. 특히 metaplasia가 일어나면서 정상 gastric mucosa에서는 보이지 않던 SF을 강하게 발현하는 cell들이 isthmus region에서 확인 되었다. 또 위암 환자의 조직에서도 Dclk1 isoform 발현의 차이를 확인해본 결과, 정상 조 직과 intestinal metaplasia (IM) 조직에서는 LF과 SF 모두 강한 발현을 보인 반면에 비해 cancer 조직에서는 LF의 발현이 소실되고 SF만 발현하는 특징을 확인하였다. 이상의 결 과는 Dclk1 isoform중 LF의 발현이 위암의 발생 과정에서 매우 중요할 것으로 사료된다.