인체 간암세포주(Hep 3B)의 증식에 미치는 amiloride의 억제효과

신채희,박기룡,최병주,서수홍,김성훈,박무인,박선자,정근옥,박건영,구자영 고신대학교 의학부 2002 高神大學校 醫學部 論文集 Vol.17 No.1

Background/Objective Cytoplasmic alkalinization induced by activation of the Na+/H+antiporter which is stimulated upon the addition of growth-promting agents, such as insulin, epidermal growth factor, phorbol ester, plays an essential role in the initiation on cell proliferation. In the present study the effects of amiloride, a specific and reversible inhibitor of Na+/H+exchanger, on the growth of human hepatocellular carcinoma cell line, Hep 3B were examined and the effects of 5-fluorouracil (5-FU) combined with amiloride were also studied to determine the role of amiloride in the treatment of hepatocellular carcinoma. Cell cycle analysis was done to examine the mechanisms for the inhibitory effects of amiloride. Methods The growth of Hep 3B cells were examined by counting cell number on two and four days treatment with 1μM, 10μM, 20μM, 40μM, 80μM, 160μM, amiloride, and 0.1㎍/㎖, 0.3㎍/㎖ 5-FU, after plating Hep 3B cells into 35-mm^(2) plastic dishes at a density of 20×10^(4) cells/dish. The reversibility of the effects of amiloride was examined on two days to eight days treatment with 20μM amiloride after seeding 4×10^(4) cells/dish Cell cycle analysis was done on the cells after four days treatment with 20μM amiloride. Results Amiloride significantly inhibited the growth of Hep 3B in a dose-dependent fashion (p<0.05). The inhibitary effect of amilride on the growth of Hep 3B cells was firstly shown at the concentration of 1μM, which is not so higher than the concentration of 0.1-0.2μM attainable by administration of usual dose of amiloride (5∼10㎎). Forty-eight percent inhibition of growth was found at an amiloride concentration of 20μM and 92% inhibition of growth was found at an amiloride concentration of 160μM after 4days treatment. The removal of amiloride by a media change after 48 hours treatment lead to significantly more growth than amiloride treated group (p<0.05). Amiloride combined with 5-FU significantly inhibited the growth of Hep 3B in a dose-dependent fashion compared to an amiloride or a 5-FU of cells in G0-G1 phase, S phase and G2-M phase was 57.9%, 20.8%, 21.3%, respectively in the amiloride group (20μM) and 38.9%, 20.8%, 40.3% in the control group, showing much higher G1 fraction in amiloride group compared to control group. Conclusions Amiloride significantly inhibited the growth of Hep 3B in dose-dependent fashion, which may be reversible. The reversibility of growth inhibition suggests that amiloride is not a non-specific cytotoxin for Hep 3B cells. Because the lowest inhibitory concentration of amiloride for the growth of Hep 3B cells in this study was 1μM, which is not so higher than the concentration of 0.1∼0.2 μM attainable by administration of usual dose of amiloride(5∼10㎎). amiloride or its analogues may be used alone or in conjunction with other modalities of therapy of hepatocellular carcinoma, for example, hepatic arterial chemoembolization of radiofrequency interstitial thermal ablation therapy. Since Hep 3B cell transition through the G1 phase was inhibited by amailoride, the inhibitory effects of amiloride on the growth of Hep 3B may be mediated in part by blocking G1-S transition. Further study is needed to clarify the effects of more potent analogues of amiloride on the growth of human hepatocellular carcinoma.

인체 간암세포주 (Hep 3B)의 증식에 미치는 amiloride의 억제효과

신채희,박기룡,최병주,서수홍,김성훈,박무인,박선자,정근옥,박건영,구자영 고신대학교(의대) 고신대학교 의과대학 학술지 2002 고신대학교 의과대학 학술지 Vol.17 No.1

Background/Objective : Cytoplasmic alkalinization induced by activation of the Na+/H+ antiporter which is stimulated upon the addition of growth-promoting agents, such as insulin, epidermal growth factor, phorbol ester, plays an essential role in the initiation on cell proliferation. In the present study the effects of amiloride, a specific and reversible inhibitor of Na+/H+ exchanger, on the growth of human hepatocellular carcinoma cell line, Hep 3B were examined and the effects of 5-fluorouracil (5-FU) combined with amiloride were also studied to determine the role of amiloride in the treatment of hepatocellular carcinoma. Cell cycle analysis was done to examine the mechanisms for the inhibitory effects of amiloride. Methods : The growth of Hep 3B cells were examined by counting cell number on two and four days treatment with 1 uM, 10 uM, 20 uM, 40 uM, 80 uM, 160 uM amiloride, and 0.1 ug/ml, 0.3 ug/ml 5-FU, after plating Hep 3B cells into 35-mm2 plastic dishes at a density of 20x10^4 cells/dish. The reversibility of the effects of amiloride was examined on two days to eight days treatment with 20uM amiloride after seeding 4x10^4 cells/dish. Cell cycle analysis was done on the cells after four days treatment with 20 uM amiloride. Results : Amiloride significantly inhibited the growth of Hep 3B in a dose-dependent fashion (p<0.05). The inhibitory effect of amiloride on the growth of Hep 3B cells was firstly shown at the concentration of 1 uM, which is not so higher than the concentration of 0.1-0.2 uM attainable by administration of usual dose of amiloride (5~10 mg). Forty-eight percent inhibition of growth was found at an amiloride concentration of 20 uM and 92% inhibition of growth was found at an amiloride concentration of 150 uM after 4 days treatment. The removal of amiloride by a media change after 48 hours treatment lead to significantly more growth than amiloride treated group (p<0.05). Amiloride combined with 5-FU significantly inhibited the growth of Hep 3B in a dose-dependent fashion compared to an amiloride or a 5-FU alone (p<0.05), which suggested additive effect of the two drugs. After four days treatment with 20 uM amiloride, the fraction of cells in G0-G1 phase, S phase and G2-M phase was 57.9%, 20.8%, 21.3%, respectively in the amiloride group (20 uM) and 38.9%, 20.8%, 40.3% in the control group, showing much higher G1 fraction in amiloride group compared to control group. Conclusions : Amiloride significantly inhibited the growth of Hep 3B in a dose-dependent fashion, which may be reversible. The reversibility of growth inhibition suggests that amiloride is not a non-specific cytotoxin for Hep 3B cells. Because the lowest inhibitory concentration of amiloride for the growth of Hep 3B cells in this study was 1 uM, which is not so higher than the concentration of 0.1~0.2 uM attainable by administration of usual dose of amiloride (5~10mg), amiloride or its analogues may be used alone or in conjunction with other modalities of therapy of hepatocellular carcinoma, for example, hepatic arterial chemoembolization or radiofrequency interstitial thermal ablation therapy. Since Hep 3B cell transition through the G1 phase was inhibited by amiloride, the inhibitory effects of amiloride on the growth of Hep 3B may be mediated in part by blocking G1-S transition. Further study is needed to clarify the effects of more potent analogues of amiloride on the growth of human hepatocellular carcinoma.

제 7 차 춘계학술대회 초록집 : 구연 ; 간세포암의 발생원인에 따른 말초혈액의 T 세포 subsets 와 자연살해세포능 ( NK cell activity ) 에 관한 연구

윤병철,신채희,박기룡,이상욱,한병훈 대한간학회 2001 Clinical and Molecular Hepatology(대한간학회지) Vol.7 No.2(S)

폐암환자의 경기관지 생검 조직에서 MAGE family gene의 발현

정만홍,임동현,강상중,강승수,신채희,장태원 대한결핵 및 호흡기학회 1999 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.89 No.0

위에 발생한 원발성 소세포암

김성훈,김지연,김현영,신채희,김동완,김기환,박현용,박무인,박선자,구자영 고신대학교(의대) 고신대학교 의과대학 학술지 2002 고신대학교 의과대학 학술지 Vol.17 No.1

Primary small cell carcinoma of the stomach is a very rare cell type in gastric cancer and an extremely aggressive tumor with grave prognosis. Because of the highly malignant potency, chemotherapy for the primary therapy of small cell carcinoma is accepted generally. We report a 44-year-old man with primary advanced gastric small cell carcinoma who respond to cisplatin and etoposide combination chemotherapy.

POEMS 증후군 1례

서수홍,최병주,노치완,신채희,장성훈,김양수,어완규,김효경 고신대학교(의대) 고신대학교 의과대학 학술지 2002 고신대학교 의과대학 학술지 Vol.17 No.1

The POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin change) is a rare multisystemic disroder of unknown pathogenesis. Although the etiology is still unclear, proinflammatory cytokines such as tumor necrosis factor-a, IL-6, and paraprotein specificity for neuroendocrine tissue have been suggested to play a causative role. We report a case of POEMS syndrome presented as progressive sensorimotor neuropathy, hepatosplenomegaly, diabetes mellitus, solitary bone plasmacytoma, hypertrichosis and hyperpigmentation. The patient was treated with local irradiation and prednisolone and showed obvious neurological and systemic improvement.

급성 심근 경색 후 사망을 예측할 수 있는 폐정맥 혈류의 도플러 심초음파 지표

노치완,주승재,최병주,서수홍,신채희,김현영,김찬욱,김성만,차태준,이재우 한국심초음파학회 2001 Journal of Cardiovascular Imaging (J Cardiovasc Im Vol.9 No.2

Background:Restrictive left ventricular (LV) filling patterns after acute myocardial infarction (AMI) predict poor prognosis. Doppler indexes of LV inflow, especially peak velocity ratio of early versus late diastolic flow (E/A) and deceleration time, can predict heart failure or death. Doppler indexes of pulmonary venous flow are also used to diagnose restrictive LV filling, but their prognostic values after AMI are not known. Methods:Doppler echocardiographic examination were performed in patients with AMI (n=122) between 7 to 10 days after attack, and followed for 30 months. Death group included 9 deaths (7.4%) during follow-up. 18 age-matched patients (control group) were selected from 70 patients without death, heart failure or readmission. Doppler echocardiographic indexes of peak systolic velocity (SV), peak diastolic velocity (DV), and peak reverse flow velocity associated with atrial contraction (AR) of pulmonary venous flow were measured by transthoracic echocardiography. Results:Death group had lower SV (46.1±6.3 vs 57.0±14.7 cm/sec;p=0.059) and SV/DV ratio (1.26±0.50 vs 1.58± 0.37 ; p=0.076). Death group had significantly more patients with SV/DV ratio less than 1.3 (67% vs 17%;p=0.026). AR was significantly different between death and control groups (29.7±7.8 vs 24.7±6.8 cm/sec;p=0.023). Death group had significantly more patients with ARgreater than 25 (78% vs 33% p=0.046). Conclusion : SV/DV ratio and AR of pulmonary venous flow predicted death after AMI. 연구배경: 급성 심근 경색 후 좌심실 이완 기능이 제한성 장애인 경우 예후가 불량하며, 좌심실 유입 혈류의 도플러 심초음파 지표 중 E/A 비와 초기 유입 혈류의 감속 시간등이 사망이나 심부전 등을 예측하는데 사용된다. 폐정맥 혈류의 도플러 심초음파 지표도 제한성 장애를 진단하는데 도움을 주나, 급성 심근 경색 후 사망을 예측할수 있는 지표인지는 아직 알려져 있지 않다. 방 법: 급성 심근 경색 후 7~10일 사이에 경흉부 심초음파를 시행 받은 122명의 환자 중 9명(7.4%)이 30개월 이내에 사망하였다. 이 9명을 사망군으로 하였다. 30개월 동안 사망, 심부전, 재입원 등의 사건이 없었던 70명의 환자 중 사망군의 연령이 보정된 18명의 대조군을 선정하여, 폐정맥 혈류의 도플러 심초음파 지표인 수축기 최대 혈류 속도(peak systolic velocity:SV), 이완기 최대 혈류 속도(peak diastolic velocity:DV),좌심방 수축에 의한 역류 혈류의 최대 속도(peak reverse flow velocity associated with atrial contraction:AR) 등을 측정하였다. 결 과: SV가 사망군에서 작은 경향이 있었다(46.1±6.3 vs57.0±14.7 cm/sec;p=0.059). SV/DV 비는 사망군에서 작았으며(1.26±0.50 vs 1.58±0.37;p=0.076),SV/DV 비가 1.3 미만인 경우가 사망군에서 67%로, 대조군의 17%에 비해서 유의하게 많았다(p=0.026). AR은 사망군에서 유의하게 높았으며(29.7±7.8 vs 24.7±6.8cm/sec;p=0.023), 그 값이 25 cm/sec 이상인 경우가 사망군에서 78%, 대조군에서 33%로 사망군에서 유의하게 많았다(p=0.046). 결 론: 폐정맥 혈류의 SV/DV 비, AR 등이 급성 심근 경색후 사망을 예측할 수 있는 지표로 생각되었다.

말초혈액에서 Tg mRNA에 대한 역전사 중합효소 연쇄 반응법의 갑상선 재발암의 분자생물학적 진단

권성일,박기룡,김현영,신채희,임영찬,최영식,박요한,이강대,장희경,이재화,염하용 대한내분비학회 2002 Endocrinology and metabolism Vol.17 No.4

연구배경: 갑상선암은 다른 조직에 발생한 암에 비해 비교적 천천히 자라므로 대부분 예후가 양호하지만, 일부에서는 주위 조직으로 침윤하거나 혹은 원격 전이로 인하여 치명적인 결과를 초래할 수 있다. 갑상선전절제술 및 131^I 제거술 후 경과 관찰시 갑상선암의 재발과 전이의 진단에 있어 131^I 스캔과 혈청 Tg의 측정이 현재 임상에서 가장 많이 이용되고 있으나 이 방법에는 여러 가지의 결점이 있다. 최근 Tg mRNA에 대한 RT-PCR법을 이용한 여러 연구결과는 131^I 스캔과 혈청 Tg 측정의 결점을 보완할 수 있는 좋은 보조적인 진단법으로 이용할 수 있을 가능성을 제시하였다. 이에 말초혈액에서 측정한 Tg mRNA에 대한 RT-PCR법이 갑상선 절제술 및 방사성요드 치료 후 갑상선암의 재발 및 전이 유무의 진단에 유용한가를 알아보고자 이 연구 시행하였다. 방법: 분화된 갑상선암으로 진단된 후 갑상선전절제술을 시행받고 방사성요드 치료를 받은 환자 중 현재까지 한차례에 이상 추적 방사성요드 전신 스캔을 시행하고 추적관찰이 가능했던 유두선암 35예, 여포선암 5예를 대상으로 연구를 시행하였다. 대상군은 131^I 스캔 소견상 음성인 군(Group Ⅰ), 잔여조직이 있는 군(Group Ⅱ), 국소전이가 있는 군(Group Ⅲ), 및 원격전이 군(Group Ⅳ)으로 구분하였다. 정상 대조군은 갑상선질환이 없는 10예의 건강인으로 하였다. 대상환자의 말초혈액을 이용한 Tg mRNA에 대해 특이적인 primer를 이용하여 RT-PCR 및 nested RT-PCR을 시행하였다. 결과: 본 연구 결과는 다음과 같다. 1) 131^I 스캔 소견상 음성인 군 21예 중 1예에서 Tg가 양성소견을 보였다. Anti Tg Ab가 양성인 4예 모두 Tg가 음성을 보였다. 잔여조직이 있거나 국소전이 및 원격전이를 보인 군 19예 중 Tg가 양성인 경우는 4예였으나, Tg mRNA는 전예에서 양성이었다. 2) 131^I 스캔에서 국소 및 원격전이 소견을 보인 8예 중 4예에서 Tg가 음성으로 131^I 스캔과 혈청 Tg 사이에 불일치 소견을 보였다. 3) 말초혈액에서 특이적인 primer를 이용하여 RT-PCR 및 nested RT-PCR을 시행한 결과 대상군 40예 및 정상 대조군 10예 모두에서 Tg mRNA가 양성을 보였다. 결론: 본 연구에서 갑상선 절제술 및 방사성요드 치료 후 갑상선암의 재발 및 전이 유무를 평가함에 있어 역전사 중합효소 연쇄 반응법을 이용한 Tg mRNA 측정의 의의는 재평가되어야 한다고 생각된다. Background: Differentiated thyroid cancer is the most common endocrine malignancy. Despite advances in the treatment of thyroid cancer, disease recurrence and metastasis may occur in as many as 20% of patients, and so continues to pose major problems in its clinical management. Serum thyroglobulin (Tg) measurements, by immunoassay, are used to detect residual or recurrent thyroid cancer following thyriod ablation. However, the usefulness of immunoassay is limited by both the requirement for thyroid hormone withdrawal, to attain optimal test sensitivity, and interference by the antithyroglobulin antibody (Anti-Tg Ab). Recent studies have reported the clinical usefulness of reverse transcription-polymerase chain reaction (RT-PCR) detection of Tg mRNA in the peripheral blood of patients with differentiated thyroid carcinomas. We performed this study to evaluated the usefulness RT-PCR of Tg mRNA in peripheral blood of patients with thyroid carcinoma following a total thyroidectomy and radioiodine ablation therapy. Methods: Forty cases that underwent a total thyroidectomy and radioiodine ablation therapy were included in this study. Of the 40 patients, 35 were papillary carcinomas and 5 were follicular carcinomas. Ten normal control subjects were also studied. Tg mRNA was extracted. Then RT-PCR and nested RT-PCR, were run with specific Tg primers. Concurrently, DNA sequencing of the isolates was carried out to prove the isolates were identical to the nucleotide sequence of the Tg. Results: The Tg was detected in 4 of 19 patients, with either a residual thyroid bed, or metastasis, on a 131^I whole body scan and in 1 of 21 patients with a negative radioiodine scan. Surprisingly, the Tg mRNA was detected in all the patients and normal controls. Conclusion: From our results we can not recommend Tg mRNA, detected by RT-PCR in peripheral blood, as a tumor marker superior to that of the Tg serum level. We consider an intensive re-evaluation of the method is required before considering its clinical applications (J Kor Soc Endocrinol 17:501∼513, 2002).

POEMS 증후군 1례

서수홍,최병주,노치완,신채희,장성훈,김양수,어완규,김효경 고신대학교 의학부 2002 高神大學校 醫學部 論文集 Vol.17 No.1

The POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin change) is a rare multisystemic disorder of unknown pathogenesis. Although the etiology is still unclear, proinflammatory cytokines such as tumor necrosis factor-α, IL-6. and paraprotein specificity for neuroendocrine tissue have been suggested to play a causative role. We report a case of POEMS syndrome presented as progressive sensorimotor neuropathy, hapatosplenomegaly, diabetes mellitus, solitary bone plasmacytoma, hypertrichosis and hyperpigmentation. The patient was treated with local irradiation and prednisolone and showed obvious neurological and systemic improvement.

위에 발생한 원발성 소세포암

김성훈,김지연,김현영,신채희,김동완,김기환,박현용,박무인,박선자,구자영 고신대학교 의학부 2002 高神大學校 醫學部 論文集 Vol.17 No.1

Primary small cell carcinoma of the stomach is a very rare cell type in gastric cancer and an extremely aggressive tumor with grave prognosis. Because of the highly malignant potency, chemotherapy for the primary therapy of small cell carcinoma is accepted generally. We report a 44-year-old man with primary advanced gastric small cell carcinoma who respond to cisplatin and etoposide combination chemotherapy.