비쑥의 휘발성물질에 의한 KB 세포의 세포사멸 유도

차정단,이생금,이명섭,차인호 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.2

Artemisia scoparia (A. scoparia), perennial herb is indigenous to Korea and has been traditionally used in liver damage. We investigated the effect of the essential oil obtained from A. scoparia on apoptosis of KB cells. Cytotoxicity and cellular DNA content were analyzed by MTT assay and flow cytometry, agarose gel electrophoresis, and Hoechst 33258 staining. The caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were estimated by Western blotting method. We found that the essential oil induced the apoptosis of the KB cells by concentrations of 0.4 to 0.2 mg/ml which was verified by DNA fragmentation, apoptotic bodies, and the sub-G0/G1 ratio. The essential oil also transient caspase-9 and caspase-3 activity and cleavage of PARP in KB cells for 24 h. The essential oil-induced apoptotic cell death was accompanied by up-regulation of Bax and down-regulation of Bcl-2. In conclusion, we demonstrated that the essential oil of A. scoparia induces apoptosis in KB cells

Induction of Apoptosis in Human Oral Epidermoid Carcinoma Cell by TheEssential Oil of Artemisia Scoparia WALDST. et KITAMURA

차정단,이생금,이명섭,차인호 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.2

Artemisia scoparia (A. scoparia), perennial herb is indigenous to Korea and has been traditionally used in liver damage. We investigated the effect of the essential oil obtained from A. scoparia on apoptosis of KB cells. Cytotoxicity and cellular DNA content were analyzed by MTT assay and flow cytometry, agarose gel electrophoresis, and Hoechst 33258 staining. The caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were estimated by Western blotting method. We found that the essential oil induced the apoptosis of the KB cells by concentrations of 0.4 to 0.2 mg/ml which was verified by DNA fragmentation, apoptotic bodies, and the sub-G0/G1 ratio. The essential oil also transient caspase-9 and caspase-3 activity and cleavage of PARP in KB cells for 24 h. The essential oil-induced apoptotic cell death was accompanied by up-regulation of Bax and down-regulation of Bcl-2. In conclusion, we demonstrated that the essential oil of A. scoparia induces apoptosis in KB cells.

국소진행된 직장암에서 Cathepsin D와 p53 발현의 임상적 의의

김준상(Jun-Sang Kim),이생금(Shengjin Lee),김진만(Jin-Man Kim),조문준(Moon-June Cho) 대한방사선종양학회 2008 Radiation Oncology Journal Vol.26 No.1

목 적: 본 연구는 수술 전 방사선화학요법을 시행 받은 국소진행된 직장암환자에서 치료 전 얻어진 직장암 생검조 직 내에서의 cathepsin D (CD) 및 p53의 발현이 갖는 예후적 인자를 포함한 임상적 의의에 대하여 알아보고자 하였 다. 대상 및 방법: 1993년 7월부터1999년 6월까지 국소진행된 직장암으로 수술 전 방사선화학요법을 시행 받은 127명 의 환자 중 근치적 수술을 시행 받았고 수술 전 생검 조직의 파라핀 블록을 얻을 수 있던 89명의 환자를 대상으로 하였다. 방사선치료는 하루 1.8 Gy씩 주 5회, 25회에 걸쳐 전 골반 부위에 45 Gy까지 조사한 후에 원발종양 부위에 만 5.4 Gy 추가 조사하였다. 화학요법은 5-Fluorouracil과 Leucovorin을 5일간 일시 정주하였고 방사선치료기간 1주 및 5주째 투여하였으며 방사선치료 종료 후 약 6주경에 근치적 절제술을 시행하였다. Mouse 단클론 항체인 CD와 p53을 이용하여 면역염색을 시행하였다. CD 및p53이 종양세포 및 종양기질세포에 10% 이상 발현 시 양성으로 판정하였다. 결 과: 종양세포내 CD 발현은 57명(64%)이 양성이었고 종양기질세포내 양성은 71명(80%)이었다. p53의 종양세포내 발현은 35명(39%)이 양성을 보였다. 성별, 환자나이, 종양의 운동성, 종양의항문 연으로부터의 거리, 임상적TNM 병기에 따라 종양세포 및 종양기질세포내 CD 발현 및 p53 발현의 통계적 차이가 없었다. 종양세포와 종양기질세포내 CD발현 사이에 양적인 상호 연관성이 있었다(p=0.01). 종양세포내 CD발현과 p53의 발현은 통계적인 상호관련성이 없었다(p=0.79). 단변량분석 및 다변량분석 상 5년 전체생존율 및 무병생존율에 대한 예후인자로서 종양세포 및 종양기질세포내 CD 발현과 p53은 통계적 유의성이 없었다. 그러나 종양세포 및 종양기질세포내 CD 발현에 따른 하위집단 생존율 분석상 종양세포 및 종양기질세포내 CD 발현이 음성인 경우가 5년 전체생존율이 75%로서 가장 높았으며 종양기질세포내에만 CD가 발현된 경우는 5년 전체생존율 29%로서 가장 낮았다(p=0.01).결 론: 수술 전 방사선화학요법을 시행 받은 국소진행된 직장암 환자에서 치료 전 직장암 생검조직 내에서의 p53발현은 유의한 생존예후인자는 아니었으며 CD가 종양세포에는 발현되지 않고 종양기질세포에만 발현 시 전체생존율에 불량한 예후인자였다. Purpose: Cathepsin D (CD) is a lysosomal acid proteinase that is related to malignant progression, invasion, and a poor prognosis in several tumors. The aim of this study was to evaluate the prognostic clinical significance of CD and p53 expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer who were treated with preoperative chemoradiation. Materials and Methods: Eighty-nine patients with locally advanced rectal cancer (cT3/T4 or N+) were included in this study. Preoperative chemoradiation consisted of a dose of 50.4 Gy of pelvic radiation and two concurrent cycles of administration of 5-fluorouracil and leucovorin. Surgery was performed six weeks after chemoradiation. CD and p53 expression in pretreatment formalin-fixed paraffin-embedded tumor biopsy specimens were assessed by immunohistochemical staining using a CD and p53 monoclonal antibodies. The threshold value for a positive stain in tumor tissue and stromal cells was 1+ intensity in 10% of the tumors or stromal cells, respectively. Results: Positive CD expression was found in 57 (64%) of the tumors and 32 (35%) of the stromal cell specimens. There was no association with CD expression of the tumor or stromal cells and patient characteristics. There was a correlation between tumor CD expression with stromal cell CD expression (p=0.01). Overexpression of p53 was not a significant prognostic factor. The 5-year overall survival (OS) and disease-free survival (DFS) rates were not different between tumor CD-negative and positive patient biopsy samples (69% vs. 65%, 60% vs. 61%, respectively). The 5-year OS rates in the tumor-negative/stromal cell-negative, tumor-negative/stromal cell-positive, tumor-positive/stromal cell-negative and tumor-positive/ stromal cell-positive biopsy samples were 75%, 28%, 62%, and 73%, respectively. Stromal cell staining only without positive tumor staining demonstrated the worst overall survival prognosis for patients (p=0.013). Conclusion: Overexpression of p53 in rectal biopy tissue was not associated with prognostic significance. In the pretreatment biopsy specimens, an exclusive increase in CD expression in stromal cells without tumor expression was related to poor overall survival in patients with locally advanced rectal cancer treated with preoperative chemoradiation.

Mutational and Loss of Heterozygosity Analysis of the p53 and PTENTumor Suppressor Genes in Breast Carcinoma

서광선,이영호,나선영,박문일,김헌수,이생금 대한병리학회 2005 Journal of Pathology and Translational Medicine Vol.39 No.5

Background : Although the genetic determinants of most sporadic breast cancers remain unknown, the understanding of the molecular and genetic events that contribute to breast carcinogenesis has been significantly advanced. We investigated the clinicopathologic significance of allelic imbalance or mutation of both p53 and PTEN tumor suppressor genes in sporadic breast carcinomas. Methods : Genomic DNA from 62 breast carcinoma cases was extracted from paraffin blocks, and PCR was performed to determine loss of heterozygosity (LOH) for DNA markers around the p53 and PTEN genes and to amplify exons 5, 6, 7, and 8 of p53 and all 9 coding axons of PTEN. Results : Somatic p53 mutations were detected in 6 (9.7%) of the 62 cases. LOH for DNA markers surrounding p53 was observed in 18 (29.0%) of the 62 cases. LOH for DNA markers surrounding PTEN was detected in 29 (46.8%) of the 62 cases. Only one case (1.6%) showed somatic PTEN mutations. Tumors with LOH on 17p or p53 mutation were large in size and negative for ER, had a high Ki-67 index, and exhibited p53 immunoreactivity (p<0.05). Tumors with LOH on 10q23 were associated with c-erbB-2 positivity (p=0.018). Conclusions : Our results indicate that LOH at 17p and/or p53 mutation is significantly associated with the aggressive pathologic parameters of breast cancer.

갑상선 유두암에서 MHC class II 항원의 발현과 임상양상과의 관계

이준철,김설영,최윤선,배연선,김윤정,이인석,권기현,나소영,구본정,김영건,노흥규,이생금,김진만,조영석,송민호 대한내분비학회 2007 Endocrinology and metabolism Vol.22 No.1

Background: Papillary thyroid carcinoma is among the most curable cancers, but some patients are at high risk for recurrence or even death. MHC antigens are essential molecules for the pathogenesis of carcinoma and also the physiologic immune responses against tumor. However, there is no data about the relationship between the expression of MHC antigens and the clinical prognosis of papillary thyroid carcinoma patients. Methods: We analyzed the relationship between the various prognostic factors and the MHC antigen expression by conducting a retrospective study of 215 patients, who had undergone thyroidectomy for papillary thyroid carcinoma between 1987 and 2003. Results: The expressions of MHC class II antigens were more frequent in papillary thyroid carcinoma than in the other thyroid diseases. Yet there was no statistically significant relationship between most of the clinicopathological factors and the expression of MHC class II antigens in papillary thyroid carcinoma patients. Interestingly, an HLA-DR expression was found in 8 (30.8%) of the 26 patients in the recurrence group and in 13 (76.5%) of the 17 patients in the non-recurrence group, and HLA-DP/DQ immunoreactivity was positive in 10 (38.5%) cases of the recurrence group and in 14 (82.4%) cases of the non-recurrence group. Conclusion: Papillary thyroid carcinoma showed a more frequent expression of MHC Class II antigens. However, the recurred papillary thyroid carcinoma showed a tendency to downregulate the expression of MHC class II antigens. Hence, the molecular mechanism for the expression of MHC class II antigens might have a role in the recurrence of papillary thyroid carcinoma. (J Kor Endocrinol Soc 22:26~34, 2007) 연구배경: 갑상선 유두암은 비교적 예후가 좋은 것으로 알려져 있으나 재발을 하거나 원격전이 및 주위조직 침습을 동반하는 경우 예후가 나쁜 것으로 알려져 있다. 그러나 현재 갑상선 유두암의 예후를 예상할 수 있는 적당한 분자표지자가 없는 상태이다. MHC 항원은 갑상선암의 발생 및 진행에 대한 면역반응에 필수적인 분자로 알려져 있다. 또한 최근 진행된 연구에 의해 MHC class II 항원이 갑상선 유두암의 발암 유전자 돌연변이에 의해 발현이 증가된다는 사실이 알려졌다. 이에 저자들은 MHC class II 항원의 발현 유무와 갑상선 유두암의 임상양상의 관련성을 평가하기 위하여 본 연구를 수행하였다. 방법: 1987년부터 2003년까지 갑상선 절제술을 시행한 215명의 환자를 대상으로 의무기록 검토를 통한 임상양상 및 다양한 예후 인자들에 대한 후향적 분석을 시행하였으며 이들의 갑상선 조직에 대한 면역조직화학적 염색을 통해 MHC class II 항원의 발현 유무를 조사하였다. 추적관찰을 통해 재발 여부를 분명하게 판정할 수 있는 43명의 갑상선 유두암 환자를 선별하여 재발군과 비재발군으로 분류하였으며 두 군 간의 MHC 항원의 발현 양상에 따른 차이를 비교하였다. 결과: MHC class II 항원은 양성 질환에 비해 갑상선암에서 발현이 증가되어 있었으며 특히 갑상선 유두암에서 통계학적으로 의미 있게 발현이 증가되어 있었다. 갑상선 유두암에서 나이를 제외한 대부분의 임상적 인자 및 병리학적 인자들이 MHC class II 항원의 발현 유무에 따른 차이를 보이지 않았다. MHC class I 항원은 재발군과 비재발군 사이에서 발현 양상에 차이가 나지 않았으나 MHC class II 항원의 경우 재발군에서 발현이 억제되어 있어 비재발군과 통계학적으로 의미 있는 차이를 보였다.