Kainate로 유발시킨 seizure 후 마우스 해마형성체에서 나타나는 c-kit와 리간드의 발현양상

흥혜남,이종환,정태진,김동호 대한해부학회 1997 Anatomy & Cell Biology Vol.30 No.1

c-kit, PDGFRA의 면역조직화학적 발현 양상과 변이 분석을 통한 위장관기질종양 진단의 접근

배한익,김정식,김재훈,오현진,서인수,김종광,강병욱,유완식,정호영 대한병리학회 2010 Journal of Pathology and Translational Medicine Vol.44 No.2

Background : Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors in the gastrointestinal tract. Recently, many methods for the diagnosis of GIST have been developed including molecular diagnosis. Methods : We selected 90 cases of GIST that had presented at Kyungpook National University Hospital between 1998 and 2007. Tissue microarrays were made using core areas of tumor tissues. Immunohistochemical staining for c-kit, protein kinase C-theta, and platelet-derived growth factor receptor alpha (PDGFRA) was done. Direct sequencing of hot spot exonal areas for c-kit and PDGFRA were done using extracted DNAs of all 90 paraffin block tissues. Results : Among the 90 cases, 83.3% (75/90) were c-kit positive, 16.6% (15/90) were c-kit negative, 93.3% (84/90) were PDGFRA positive, and 6.6% (6/90) cases were PDGFRA negative. Fifteen cases of c-kit negative GIST included 1 case of PDGFRA negative and 5 cases of PDGFRA negative GIST were ckit positive. The one case in which both c-kit and PDGFRA were negative, showed a c-kit mutation in exon 11. Conclusions : Combined immunohistochemical staining of c-kit, discovered on GIST 1 (DOG1) and PDGFRA is helpful for the diagnosis of GIST. When all staining tests are negative for immunoreactivity, c-kit mutation analysis for exon 11, 9 should be done. Genotyping of kit and PDGFRA do not need to be examined initially, if it is only for the diagnosis of GIST.

비인강암에서 c-kit 단백의 발현 및 유전자 변이의 임상적 의미

강일규,이승훈,정광윤,우정수,백승국,조승현,이세우,권순영,김인선 대한이비인후과학회 2007 대한이비인후과학회지 두경부외과학 Vol.50 No.6

Background and Objectives:The proto-oncogene c-kit is a receptor tyrosine kinase recognized to initiate esential signal tra-nsduction pathways that transmit biological signals for cellular proliferation, diferentiation and metastasis. Aberrant expression -ing aditional groups of tumors that may use the stem cell factor/c-kit pathway, we investigated the expresion of c-kit in nasoph-aryngeal carcinoma. Subjects and Method:In this retrospective study, imunohistochemical stains for c-kit were performed on for-malin-fixed parafin embedded sections from 20 patients with nasopharyngeal carcinoma who were treated from 1996 to 2004. Gene mutation was analyzed by PCR-SSCP and direct sequencing. Results:c-kit over-expression was found in 65% (13/20) of patients. Eight of the 13 samples (61.5%) exhibited strongly positive immunoreactivity for c-kit protein (staining of >50% of the tumor cells). c-kit gene mutation was found in 4 of 20 cases by the PCR-SSCP method. Conclusion:c-kit protein over-expre-sion is found in 65% of the nasopharyngeal carcinoma patients. c-kit expression is corelated with c-kit DNA mutations and naso-pharyngeal carcinoma may be potential targets for treatment with imatinib mesylate. (Korean J Otolaryngol 207 ;50 :519-24)

기니이픽 장관의 c-Kit 및 NK 1R 면역반응 세포구조에 대한 공초점 주사현미경적 연구

장인엽,김종중,문정석,김현곤,박찬국,전제열,전규배,조철희,유호진 조선대학교 2001 The Medical Journal of Chosun University Vol.26 No.1

Background and Objectives: Immunolabelling of interstitial Cajal(IC) cells in the intestinal wall has recently been developed by using a specific marker, the anti-c-Kit antibody. Substance-P is a well-known neurotransmitter in the gastro-intestinal tract. Since the gastro-intestinal wall structures have already been well documented in the guinea pig, immunohistochemistry was done for the c-Kit-positive IC network and substance-P receptor(NK1R) in an attempt to provide a morphological basis for the mechanism regulating gastro-intestinal movement. Materials and Methods: Cryosection and whole-mount preparations of guinea pig small intestine and colon were single and double immunolabelled using the anti-c-Kit and NK1R antibodies. Immunolabelled specimens were observed under a confocal laser scanning microscope. Results : According to a three dimensional reconstruction study, it was found that (1) the c-Kit-positive celluar networks were widely distributed in the intestinal wall, (2) c-Kit-positive celluar networks encircled the ganlion, with strands in reticular configurations, and (3) the c-Kit-positive cells showed colocalization with NK1R in circular muscle(CM), not myenteric plexus(MY). Conclusion: The charateristic profiles of IC containing c-Kit-positive celluar networks and the relationship between c-Kit-positive and NK1R-positive structures provide a morphological basis upon the mechanism regulating gastro-intestinal motility.

ACK2로 c-kit의 기능이 저해된 발생신경계에서 나타나는 c-kit 수용체와 리간드의 발현

홍혜남,이종환,김동호 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.4

출생 후 소뇌 GABA성 신경세포의 성장과 분화에 기여하는 줄기세포인자와 c-kit 수용체의 역할

원유진(You Jin Won),이종환(Jong Hwan Lee),임진옥(Jin Ok Im),윤승용(Seung Yong Yun),황승준(Seung Jun Hwang),김동호(Donghou Kim),홍혜남(Hea Nam Hong) 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.5

출생 후 소뇌의 분자층에 있는 억제성 사이신경세포에서는 c-Kit (c-kit gene의 단백질 산물)이 발현되고 이 세포들과 억제성 신경연접관계에 있는 조롱박세포에서는 줄기세포인자(Stem Cell Factor; SCF)가 발현된다는 사실은 알려져 있으나 c-Kit과 SCF가 출생 후 소뇌세포의 성장과 분화에 어떤 역할을 하는지에 관해서는 거의 밝혀진 바가 없다. 본 연구는 출생 후 마우스의 소뇌세포 배양 시 c-Kit 항체 처리로 c-kit 수용체의 기능을 억제하거나 SCF를 처리하는 방법으로 소뇌세포의 출생 후 발달과정에 기여하는 c-Kit과 SCF의 역할을 관찰하였다. 그 결과 SCF는 소뇌세포의 생존율을 비롯하여 GABA성 신경세포의 Glutamic Acid Decarboxylase 65/67 (GAD65/67) 합성을 증가시켰을 뿐만 아니라 소뇌 억제성 사이신경세포와 조롱박 세포의 성장을 돕고 신경돌기의 분화를 촉진시켰다. 한편 c-Kit 항체의 처리로 c-kit 수용체의 기능이 억제되면 소뇌세포의 생존율 저하와 GABA성 신경세포에서의 현저한 GAD65/67 발현양 감소를 나타내었으며, SCF와 c-Kit 항체가 복합처리 되면 c-Kit 항체로 인하여 SCF의 활성이 저해되었으므로 이 두 인자가 서로 상호작용하고 있음을 알 수 있었다. 또한 c-Kit 항체는 억제성 사이신경세포와 조롱박 세포의 성장과 신경돌기의 신장 및 분화를 강력히 억제하였다. 따라서 SCF와 c-kit 수용체는 억제성 사이신경세포와 조롱박 세포의 출생후 정상적인 성장과 분화과정에 필수적인 역할을 하고 있는 것으로 추정할 수 있다. Evidence that Stem cell factor (SCF) and c-Kit receptor tyrosine kinase are expressed in the cerebellum during postnatal development, suggests a possible contribution of the SCF/Kit signaling pathway in the cerebellar development. In the present study, we prepared cerebellar cultures from C57Bl/6J mouse at postnatal day 1and 7 to investigate the role of c-kit receptor and SCF in regulation of growth and differentiation in the postnatal cerebellar GABAergic cells. SCF increased the number of survival cerebellar cells and density of glutamic acid decarboxylase 65/67 (GAD65/67) and calbindin D-28K expression in the immunoblot analysis. SCF also improved the neurite extension of the interneuron neuritis and dendritogenesis of Purkinje cells. Treatment with c-Kit antibody accelerated cellular loss in serum-free media and decreased the growth ability and dendritogenesis of Purkinje cells and cerebellar inhibitory interneurons. Our data suggest that SCF and c-kit receptor are required for the normal growth of postnatal cerebellum and a possible involvement of functional regulation through the SCF/c-kit receptor pathways in the postnatal cerebellar development.

소 c-KIT Receptor 유전자의 다형성에 관한 연구

장요순,김태헌,윤두학,박응우,이혜원,이학교,정일정 한국동물자원과학회 2002 한국축산학회지 Vol.44 No.6

소의 흰 반점 관련 후보유전자로 c-KIT receptor 유전자를 선정하여, c-KIT receptor 유전자내의 변이를 탐색하고 변이가 흰반점 표현형과 연관성이 있는지를 분석하였다. 한우, Angus, Brown Swiss, Charolais, Hereford, Holstein, Limousin 및 Simmental 등 8개 품종의 DNA 시료를 사용하여 c-KIT receptor 유전자의 intron 6번 영역에서 다형성을 조사하고 분석하였다. c-KIT receptor 유전자의 intron 6번 영역에서는 4개의 염기치환이 발견되어, MspⅠ, BsrBⅠ 및 NdeⅠ 제한효소를 이용하여 PCR-RFLP 분석을 실시하였다. Intron 6번을 포함하는 영역의 PCR 산물 크기는 2,440 bp 이었다. MspⅠ다형성은 PCR-RFLP 분석 결과 3개의 대립유전자가 존재하였으며, 한우품종에서는 3개의 대립유전자 모두가 발견되었고, CC 형태이 유전자형을 제외한 5개의 유전자형 (AA, AB, AC, BC 및 BB)을 확인하였다. Angus, Brown Swiss, Hereford, Holstein 및 Simmental 품종에서는 A 대립유전자만을 갖는 것으로 조사되었고, 한우는 44%만 AA 유전자형을 나타내었다. BsrBⅠ 다형성은 2개의 대립유전자로서 3개의 유전자형이 나타나는 것을 확인하였으며, Charolais 및 Hereford 품종이 다른 소 품종에 비하여 A 대립유전자의 빈도가 높게 나타났다. NdeⅠ 다형성을 분석한 결과 Brown Swiss 품종에서는 NdeⅠ에 의해 절단되는 형태인 A 대립유전자만 관찰되었으며, Holstein 품종은 92%, Simmental 품종은 72%가 절단되는 형태를 나타내어, 모색이 흰색을 띠는 소 품종에서 절단되는 형태가 많았다. 소 c-KIT receptor 유전자의 intron 6번 영역에서 확인된 4개의 염기치환은 품종에 따라 다른 빈도를 보였으나, 이들 염기치환과 흰 반점과의 연관성에 대한 증거는 발견하지 못하였다. 그러므로 소의 흰 반점과 c-KIT receptor 유전자 내의 변이와의 관련성은 다른 영역에 대한 추가적인 분석과, 이미 보고된 다른 모색관련 유전자의 다형성과의 연관성 분석 등과 같은 연구가 필요한 것으로 판단된다. We considered KIT gene as a candidate gene for the white-spotting pattern in cattle. This study was carried out to detect genetic variation of c-KIT receptor gene and to investigate association between the mutation and the white-spotting pattern in cattle. PCR-RFLP analysis within intron 6 of c-KIT receptor gene were performed with 8 cattle breeds including Hanwoo, Angus, Brown Swiss, Charolais, Hereford, Holstein, Limousin and Simmental. When PCR product of approximately 2,440 bp including intron 6 of c-KIT receptor gene was sequenced, four nucleotide substitutions were found within intron 6 of the bovine c-KIT receptor gene. In PCR-RFLP analysis, three alleles (A, B and C), two alleles (A and B) and two alleles (A and B) at each locus were identified by Msp Ⅰ, BsrBⅠ and NdeⅠ, respectively. Although frequencies of allele at each locus were different among cattle breeds, we could not get any evidence related with white or white spotting phenotypes in these mutations on intron 6 of c-KIT receptor gene. However, we can not entirely exclude the possibility that c-KIT receptor gene is responsible for white spotting phenotype in cattle. Thus, further studies need to detect other mutations in c-KIT receptor gene and to test association of those mutations and coat color phenotypes in cattle.

Estrogen-induced transcription factor EGR1 regulates c-Kit transcription in the mouse uterus to maintain uterine receptivity for embryo implantation

Park, Mira,Kim, Hye-Ryun,Kim, Yeon Sun,Yang, Seung Chel,Yoon, Jung Ah,Lyu, Sang Woo,Lim, Hyunjung Jade,Hong, Seok-Ho,Song, Haengseok North-Holland 2018 Molecular and cellular endocrinology Vol.470 No.-

<P><B>Abstract</B></P> <P>Early growth response 1 (Egr1) is a key transcription factor that mediates the action of estrogen (E<SUB>2</SUB>) to establish uterine receptivity for embryo implantation. However, few direct target genes of EGR1 have been identified in the uterus. Here, we demonstrated that E<SUB>2</SUB> induced EGR1-regulated transcription of c-Kit, which plays a crucial role in cell fate decisions. Spatiotemporal expression of c-Kit followed that of EGR1 in uteri of ovariectomized mice at various time points after E<SUB>2</SUB> treatment. E<SUB>2</SUB> activated ERK1/2 and p38 to induce EGR1, which then activated c-Kit expression in the uterus. EGR1 transfection produced rapid and transient induction of c-KIT in a time- and dose-dependent manner. Furthermore, luciferase assays to measure c-Kit promoter activity confirmed that a functional EGR1 binding site(s) (EBS) was located within −1 kb of the c-Kit promoter. Site-directed mutagenesis and chromatin immunoprecipitation-PCR for three putative EBS within −1 kb demonstrated that the EBS at -818/-805 was critical for EGR1-dependent c-Kit transcription. c-Kit expression was significantly increased in the uterus on day 4 and administration of Masitinib, a c-Kit inhibitor, effectively interfered with embryo implantation. Collectively, our results showed that estrogen induces transcription factor EGR1 to regulate c-Kit transcription for uterine receptivity for embryo implantation in the mouse uterus.</P> <P><B>Highlights</B></P> <P> <UL> <LI> c-Kit expression was significantly reduced in uteri of Egr1 (−/−) mice. </LI> <LI> Spatiotemporal expression patterns of c-Kit coincided with those of Egr1 in the uterus after E<SUB>2</SUB> treatment. </LI> <LI> E<SUB>2</SUB>-ER-dependent activation of the ERK1/2 and p38 MAPK-EGR1 pathway regulated c-Kit induction in the uterus. </LI> <LI> The EBS at −818/-805 was critical for EGR1-dependent activation of c-Kit transcription. </LI> <LI> c-Kit expression was significantly increased in mouse uteri receptive for embryo implantation. </LI> </UL> </P>

급성 백혈병세포에서 c-kit 표현에 관한 연구(I)

민유홍,장길진,라선영,이선주,한지숙,고윤웅 大韓血液學會 1993 大韓血液學會誌 Vol.28 No.2

Radotinib induces high cytotoxicity in c-KIT positive acute myeloid leukemia cells

Heo, S.K.,Noh, E.K.,Kim, J.Y.,Jo, J.C.,Choi, Y.,Koh, S.,Baek, J.H.,Min, Y.J.,Kim, H. North-Holland 2017 European journal of pharmacology Vol.804 No.-

<P>Previously, we reported that radotinib, a BCR-ABL1 tyrosine kinase inhibitor, induced cytotoxicity in acute myeloid leukemia (AML) cells. However, the effects of radotinib in the subpopulation of c-KIT-positive AML cells were unclear. We observed that low-concentration radotinib had more potent cytotoxicity in c-KIT-positive cells than c-KIT-negative cells from AML patients. To address this issue, cell lines with high c-KIT expression, HEL92.1.7, and moderate c-KIT expression, H209, were selected. HEL92.1.7 cells were grouped into intermediate and high c-HIT expression populations. The cytotoxicity of radotinib against the HEL92.1.7 cell population with intermediate c-HIT expression was not different from that of the population with high c-KIT expression. When H209 cells were grouped into c-KIT expression-negative and c-HIT expression-positive populations, radotinib induced cytotoxicity in the c-KIT-positive population, but not the c-KIT-negative population. Thus, radotinib induces cytotoxicity in c-KIT-positive cells, regardless of the c-KIT expression intensity. Therefore, radotinib induces significant cytotoxicity in c-KIT-positive AML cells, suggesting that radotinib is a potential target agent for the treatment of c-KIT-positive malignancies including AML.</P>