• Title/Summary/Keyword: 두경부 편평상피세포암 세포주

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Beta-4 Integrin Transfection, Cloning and Functional Assay in Squamous Cell Carcinoma (Beta-4 Integrin 유전자 주입, 클로닝과 편평상피암에서의 Beta-4 Integrin 기능에 관한 연구)

  • Kim Young-Min;Carey Tomas E.
    • Korean Journal of Head & Neck Oncology
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    • v.13 no.2
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    • pp.169-179
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    • 1997
  • 서론 : Laminin의 수용기로 알려진 Integrin $\alpha6\beta4$의 세포내 표현 정도는 편평상피암을 위시한 여러 악성종양의 전이능력 및 예후와 밀접한 상관관계가 없다고 알려져 있다. 이 Integrin은 Laminin과 같은 세포와 리간드와 결합하면 상피세포의 기저막 지주 구조물인 hemidesmosome의 세포체질 요소(cytoskeletal element)와 연관되어 그 결과 세포의 기저막과 세포내 케라틴을 연결하는 역할을 한다. Integrin $\alpha6\beta4$는 구조적으로 다른 많은 integrin들과 달리 $\beta$4의 세포질내 영역(cytoplasmic domain)이 특징적으로 크다. 이 세포질내 영역 $\beta$4 integrin의 기능은 아직 밝혀지지 않고 있으나 아마 세포 성장의 신호전달 및 악성종양의 특징인 침윤 전이에 관련할 것으로 보아지고 있다. 재료 및 방법: 저자들은 우선 $\beta$4 integrin의 wild type s-DNA와 $\beta$4 세포질내 영역(cytoplasmic domain) 및 $\beta$4의 tyrosine 인산화 반응 부위가 각각 결손된 c-DNA를 PCR을 통하여 합성하여 pRc/CMV 벡터에 삽입한 후 원래 $\beta$4 integrin의 발현이 결집된 인간 방광암 세포에 Calcium phosphate precipitation 방법으로 주입(transfection)시켜 형질변환된 세포를 면역형광법, Flow cytometry 및 Immunoprecipitation 방법으로 클로닝하여 wild type $\beta$4-full length(Clone FL), truncated $\beta$4-cytoplasmic domain(C1one CD), 및 mutated $\beta$4-tyrosine phosphorylation site (Clone M)을 얻었다. 암 세포의 부착 및 침투 능력의 기능적 연구로 모노 클로날 항체와 fibronectin, laminin, Matrigel을 단백질 기질로 사용하였으며 결과 비교를 위하여 pRc/CMV 벡터만 주입시켰던 클로운과 방광암 세포주를 $\beta$4 integrin 음성 대조군으로 또한 이 Integrin의 높은 발현을 보이는 두경부 편평상피암 세포주를 양성 대조군으로 이용하였다. 결과 : 세포부착능력에 있어서 온전한 $\beta$4 cytoplasmic domain이 존재하는 클로운이 laminin에 강한 부착능력을 보였으나 fibronectin의 부착정도는 $\beta$4 integrin의 표현정도와 관계없이 모든 클로운에서 비슷하였다. Matrigel을 투과하는 암세포 침윤 능력에서는 $\beta$4 integrin의 표현이 존재하는 클로운들이 투과 능력이 높았으나 세포외 리간드가 없는 control membrane을 사용하였을 때와 비교하여 투과능력의 차이를 보이지 않았다. 결론 : 유전자 주입(transfection) 방법으로 integrin의 다양한 클로운의 합성이 가능하여 이 Integrin의 암 세포의 부착 및 침투 능력에서의 기능을 규명 할 수 있게 한다. $\beta$4 integrin은 편평상피 암세포의 부착에 있어서 세포외 리간드 laminin과 특이 결합하여 부착 능력을 높이는 중요한 역할을 하며 편평상피 암세포의 침투에 있어서는 $\beta$4 integrin의 표현이 침투 능력을 높이는 역할을 하나 이때에는 laminin과 같은 리간드와의 특이 결합에 의존하지는 않는 것으로 사료된다.

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Activation and Abnormalities of Cell Cycle Regulating Factor in Head and Neck Squamous Cell Carcinoma Cell Lines: Abnormal Expression of CDKN2 Gene in Laryngeal Squamous Cell Carcinoma (두경부 편평상피세포암 세포주에서 세포주기조절인자의 활성 및 이상 : 후두편평상피세포암에서 종양억제유전자 CDKN2 유전자의 발현이상)

  • Song, Si-Youn;Han, Tae-Hee;Bai, Chang-Hoon;Kim, Yong-Dae;Song, Kei-Won
    • Journal of Yeungnam Medical Science
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    • v.22 no.2
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    • pp.166-182
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    • 2005
  • Background: Cyclin-dependent kinase (CDK) inhibitors are family of molecules that regulate the cell cycle. The CDKN2, a CDK4 inhibitor, also called p16, has been implicated in human tumorigenesis. The CDKN2 inhibits the cyclin/CDK complexes which regulate the transition from G1 to S phase of cell cycle. There is a previous report that homozygous deletion of CDKN2 region on chromosome 9p21 was detected frequently in astrocytoma, glioma and osteosarcoma, less frequently in lung cancer, leukemia and ovarian cancer, but not detected in colon cancer and neuroblastoma. However, little is known about the relationship between CDKN2 and laryngeal cancer. Therefore this study was initiated to investigate the role of CDKN2 in human laryngeal squamous cell carcinoma development.1) Materials and methods: We used 5 human laryngeal carcinoma cell lines whether they have deletions or losses of CDKN2 gene expression by DNA-PCR or RT-PCR, respectively. We examined 8 fresh frozen human laryngeal cancer tissues to detect the loss of heterozygosity (LOH) of CDKN2. PCR was performed by using microsatellite markers of short arm of human chromosome 9 (D9S126, D9S144, D9S156, D9S161, D9S162, D9S166, D9S171, D9S200 and D9SIFNA). For informative cases, allelic loss was scored if the signal of one allele was significantly decreased in tumor DNA when compared to the same allele in normal DNA. Results: The CDKN2 DNA deletion was observed in 3 cell lines. The CDKN2 mRNA expression was observed in only one cell line, which was very weak. LOH was detected in 7 cases (87.5%). Conclusion: These results suggest that CDKN2 plays a role in the carcinogenesis of human laryngeal squamous cell carcinoma.

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The Modulation of Squamous Cell Differentiation by Retinoids in Human Squamous Cell Carcinoma Xenografts (Nude Mouse 에 이종이식한 두경부 편평상피세포암의 분화에 대한 Retinoids의 작용)

  • Kim, Sang-Yoon;Yoo, Seung-Joo;Yoo, Keun-Sik;Joo, Joon-Bum;Choi, Doo-Yung;Nam, Soon-Yuhl
    • Korean Journal of Bronchoesophagology
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    • v.5 no.2
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    • pp.119-126
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    • 1999
  • Objectives : To analyze the effect of retinoids on the differentiation in HNSCC xenografts. Materials and Methods : RA (20mg/kg) or 13-cis-RA (60mg/kg) was orally administered once in a day for 30 days in the xenograft model we prepared using athymic nude mice with AMCHN-4 and -6. We carried out H & E staining and immunohistochemical staining with the monoclonal antibody against involucrin and cytokeratin 10. Results : Both RA and 13-cis-RA were found to suppress the differentiation of AMC-HN-4. Interestingly, RA enhanced the differentiation of AMC-HN-6, although 13-cis RA did not exhibit any effect on the differentiation. These results suggest that in vivo effect of retinoids on the HNSCC growth and differentiation might be various. Retinoids-induced P450 in AMC-HN-6 might be one of the mechanisms to explain the reason why the retinoids exhibit various functions in the HNSCC.

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Radiosensitization of Cis-Platimum in the Treatment of Advanced Head and Neck Squamous Cell Carcinoma (국소 진행된 두경부편평 상피암에 대한 CIS-PLATINUM과 방사선치료의 동시 병행요법)

  • Chang, Hye-Sook
    • Radiation Oncology Journal
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    • v.10 no.1
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    • pp.27-34
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    • 1992
  • Cis-Platinum (DDP) was utilized as a radiosensitizer in a pilot study for stage III and IV squamous cell carcinoma between 1984-1987, and DDP 20 $mg/M^2$/day was administered for 4 days at 3 week interval with concurrent radiotherapy. This study consisted of three phases: cytoreduction phase, eradicative treatment phase and adjuvant phase. Total 59 patients were subjected to evaluate a tumor response and its toxicity. During the eradicative phase,27 patients underwent surgery (group I ), 29 patients were treated with radiotherapy only (group II) and 3 patients did not complete the second phase of therapy. At the cytoreduction phase, $95\%$ response rate with complete response (CR) $47.5\%$ and partial response (PR) $47.5\%$ was observed. Complete tumor clearance (CTC) rate following 2nd phase of therapy was $84\%$ (47/56) with 26/27($96\%$) in group I achieved CTC with surgery and 21/29 ($72\%$) patients In group II achieved CTC following 2nd phase. $67\%$ of primary lesions and $70\%$ of nodal diseases in group I showed no tumor in the surgical specimen. $34\%$ of patiets who achieved CTC at 2nd phase developed recurrence and median time to recur was 8 months. Actuarial disease free survival at 4 years was $59\%$ and $51\%$(24/27) of patients who achieved CTC at 2nd phase were alive without any evidence of disease at median follow-up 31 months (range, 10-48 months). There was no significant difference in overall and disease free survival between group I and II between CR and PR group following 1st Phase. Only significant Prognostic factor in this study was the complete tumor clearance following 2nd phase theapy. In general, toxicity was not excessive. Author concludes that this study confirmed the significant radiosensitizing effect of DDP with the acceptable toxicity and warrant the prospective study to determine optimum scheduling for DDP and radiotherapy which maximizes the therapeutic gain.

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Results of Curative Treatment for Cancer of the Tonsil (편도암의 근치적 치료 결과)

  • Park, Won;Ahn, Yong-Chan;Lim, Do-Hoon;Baek, Chung-Whan;Son, Young-Ik;Park, Keun-Chil;Kim, Kyoung-Ju;Lee, Jeung-Eun;Kang, Min-Kyu;Park, Young-Je;Nam, Hee-Rim;Huh, Seung-Jae
    • Radiation Oncology Journal
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    • v.21 no.4
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    • pp.261-268
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    • 2003
  • Purpose: To report the results of curative treatment for patients with tonsil cancer by retrospective analysis. Materials and Methods: From Jan. 1995 till Dec. 2000, 27 patients with squamous cell carcinoma of the tonsil received curative treatment at Samsung Medical Center. Therapeutic decision was made through multidisciplinary conference, and curative radiation therapy was favored when, (1) the patient's condition was not fit for general anesthesia and surgery, (2) the patient refused surgery, (3) complete resection was presumed impossible, or (4) too severe disability was expected after surgery. Surgery was the main local modality in 17 patients (S$\pm$RT group), and radiation therapy in 10 (RT$\pm$CT group). The median follow-up period was 41 months. Results: AJCC stages were I/II in four, III in two, and IV in 21 patients. The 5-year disease-free survival rate was 73.3$\%$ in all patients, 70.6$\%$ in the S$\pm$RT group, and 77.8$\%$ in the RT$\pm$CT group. Treatment failure occurred in seven patients, all with stage III/IV, and all the failures occurred within 24 months of the start of treatment. Five patients among the S$\pm$CT group developed treatment failures; 2 local, 2 regional, and 1 distant (crude rate=29.4$\%$). Two patients among the RT$\pm$CT group developed failures; 1 synchronous local and regional, and 1 distant (crude rate=20.0$\%$). The 5-year overall survival rate was 77.0$\%$ in all patients, 80.9$\%$ in the S$\pm$RT group, and 70.0$\%$ in the RT$\pm$CT group. Conclusion: We could achieve favorable results that were comparable to previously reported data with respect to both the rates of local control and of survival by applying S$\pm$RT and RT$\pm$CT. RT$\pm$CT is judged to be an alternative option that can avoid the functional disability after surgical resection.