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Graves병 환자의 HLA-DQ β 유전자의 제한효소 분절 길이의 다형성 : Restriction Fragment Length Polymorphism, RFLP
양인명,팽정령,김성운,김진우,김영설,김광원,최영길 경희대학교 유전공학연구소 1990 遺傳工學論文集 Vol.2 No.-
The distributions of HLA-DR and DQ antigen were compared between 40 patients with, Graves' disease and 24 normal controls. The frequency of HLA-DR 7 among 16 types of HLA-DR was significantly lower in the patient group. And the frequency of HLA-DQw4 among 6 types of HLA -DQ was significantly higher in the patients group. We compared the restriction fragments of HLA-DQ β DNA digested with Bam HI and Taq I and hybridized with DQ β cDNA between 10 patients and 10 normal controls who were homozygotes of HLA-DQw1 or DQw6. There was no band showing significant difference in the frequency. These data suggest that the pattern of linkage between HLA class Ⅱ antigens and the disease susceptibility in Korean is different from that in Caucacian, and a gene responsible for disease suceptibility is not located on DQ βgene (J Kor Soc Endocrinol 5:29-36, 1990).
Craves병 환자에서 백혈구와 갑상선 조직의 HLA-DR β 유전자 부위의 비교
양인명,우정택,팽정령,서광식,김성운,김영설,김광원,최영길 경희대학교 유전공학연구소 1989 遺傳工學論文集 Vol.1 No.-
정상 갑상선세포에서는 HLA-DR 항원이 발현되지 않았으나, 그레이브스병 환자의 갑상선세포 표면에는 HLA-DR 항원이 발현됨이 보고되어, 이러한 현상은 이 질환의 자가면역 기전에 중요한 역할을 하고 있을 것으로 생각되고 있다. 한편 최근 DR이나 DQ 유전자의 상부 159kb 이내에는 이들의 발현을 조절하는 유전자가 존재함이 알려져 있고, 이 부위의 구조적인 변화로 인하여 DR DQ 유전자의 발현을 유도하는 여러 가지 핵내 인자들과 interferon-r 와 같은 외부인 자들이 보고되고 있다. 그러나 그레이브스병에서 이들 유전자 부위의 구조적인 변화에 관해서는 아직 보고가 없다. 이에 본 연구자 등은 이러한 가능성 여부를 규명하고자 2명의 전형적인 그레이브스병 환자의 수술 우 얻어낸 갑상선 조직에서 RNA를 분리한 우 DR β유전자를 소식자로 northem blotting을 하여 mRNA의 발현을 관찰하였으며, 말초혈액 백혈구와 갑상선 조직으로부터 분리된 DNA를 EcoRI BamHI. HindⅢ PvuⅡ TaqI, PstI등의 6가지 제한효소로 소화한 후 DR β유전자를 소식자로 하여 RFLP 양상을 비교한 결과, 환자 모두에서 mRNA의 발현이 관찰되었으나, 환자 모두에서 6가지 제한효소에 의한 RFLP 양상이 동일하였다. 이러한 결과는 Graves병 환자의 DR 유전자의 발현에 있어서 이 유전자 부위의 구조적인 변화가 관여할 가능성이 적음을 시사하는 사실이라고 사료되나 향후 더 많은 예와 더 많은 제한효소를 이용한 주시가 필요할 것이다. The requirement for major histocompatibility antigen class Ⅱ molecules in the recognition of antigen by helper T cells suggests that the expression of class Ⅱ antigen may be important in the initiation and prolongation of immunopathology. HLA class Ⅱ antigenes are expressed on the surface of thyrocytes of the patients with graves disease. The increased expression of class Ⅱgene can be induced by trans acting factor such as interferon However the possibility of rearrangement of their regulatory genes has not been explored so far. We studied the mRNA expression in the thyrocytes of 2 patients with Graves' disease and compared the restriction fragment length polymorphism (RFLP) between thyroid and peripheral leukocyte DNA. The prominent expression of mRNA was observed in the thyroid tissues of all the two patients. But we did not find any difference in RFLP pattern in both patients. These results suggest the possibility that the rearrangement of the regulatory gene located in the upstream of DR- β gene can be a role in expression of DR antigen is less likely.
Gsa 유전자의 돌연변이가 쥐의 소마토스타틴 수용체 유전자의 발현에 미치는 효과
양인명,박승준,장호강,이군자,오승준,김성운,김진우,김영설,최영길 대한내분비학회 1999 Endocrinology and metabolism Vol.14 No.4
Background: The growth hormone (GH)-secreting pituitary adenoma with mutation of Gsa gene (gsp oncogene) is known to show a higher response to somatostatin. The mechanism for the higher response is unclear. We previously demonstrated that transcription of the rat somatostatin receptor (SSTR) gene was increased by cAMP. Therefore, we investigated whether the mutation of Gsa gene can increase SSTR gene expression. Methods: The mutant Gsa expressing plasmid of which arginine of codon 201was replaced with histidine was transfected transiently and permanently into GH3 cells. cAMP and rat GH were measured by radioimmunoassay. mRNA expression of the rat SSTR2 was determined by a semiquantitative RT-PCR. Results: The intracellular cAMP production was increased by 1.8 folds in the transiently transfected cells and by 1.5 folds in permanently transfected cells compared to those in the cells transfected with the vehicle plasmid, plasmid expressing the wild type or the plasmid expressing the silent mutant of codon 226. The transcriptional activation by cAMP response element of somatostatin.gene was also increased by 2 folds 24 hours after transient transfection and by 3.5 folds in the permanently transfected cells. The transcriptional activation was not enhanced by forskolin in the transiently transfected cells, whereas it was more remarkably inhibited by somatostatin compared to that in the other transfected cells. The expression of SSTR2 was increased by 1.8 folds 24 hours after transfection in the transiently transfected cells and by 3 folds in the permanently transfected cells, and it was not enhanced by forskolin and isobutylmethylxanthine. The secretion of GH from the transiently transfected cells was not significantly higher than that from the wild type-expressing cells, but it was higher in the permanently transfected cells. The suppression of GH by somatostatin was more prominent in the permanently transfected cells compared to the other transfected cells. Conclusion: This study suggests that a higher expression of SSTR induced by the mutant Gsa may be a mechanism by which gsp-positive adenomas showed a higher response to somatostatin (J Kor Soc Endocrinol 14:657-666, 1999).