2000 Fiscal Year Annual Research Report

肺癌細胞のテロメラーゼ活性・遺伝子変異と化学療法反応性の検討

Research Project

Project/Area Number	11670579
Research Institution	HIROSHIMA UNIVERSITY
Principal Investigator	檜山桂子広島大学, 医学部, 助手 (60253069)
Co-Investigator(Kenkyū-buntansha)	礒部威広島大学, 医学部・附属病院, 助手 (70284198) 石岡伸一広島大学, 医学部, 講師 (10191868)
Keywords	肺癌 / テロメラーゼ / hTERT / 化学療法 / 細胞診 / 予後 / alternative splicing / 遺伝子
Research Abstract	肺癌非手術検体における遺伝子変異・テロメラーゼ活性を測定し、それらと、予後・抗癌剤反応性との関係を検討することを試みた。肺癌非手術症例の経気管支鏡的擦過細胞診検体等について、細胞診用スライド作製後残った細胞から蛋白を抽出し、核ペレットからはDNAを抽出した。蛋白溶液のテロメラーゼ活性をTRAP法にて測定したが、細胞診検体中の肺癌細胞が占める割合は種々で、かつ、非小細胞癌では元来テロメラーゼ活性レベルが低い症例が過半数を占めるため、検出率が約40%と低いのみならず、活性レベルは混入する正常細胞の割合に左右された。そこで、感度が高く正常細胞が多数混入しても影響されない方法の確立をめざした。まず、テロメラーゼ蛋白成分のhTERT mRNAを定量的RT-PCR法で測定した。alternative spllcingされていない全長mRNAの発現量とテロメラーゼ活性はある程度相関したが、必ずしもテロメラーゼ活性レベルを正しく反映するものではなく、mRNA発現がありながら活性が認められない症例があった。そこで、hTERT抗体を用いた免疫染色を試みたところ、テロメラーゼ高活性例は例外なく染色される癌細胞の割合が高く、低活性例では染色されない癌細胞の比率が高かった。すなわち、細胞診検体のように正常細胞が多数混入する検体であっても、その中の癌細胞についてhTERT陽性細胞:陰性細胞比で、もともとの癌のテロメラーゼ活性レベルが推定できると考えられた。さらに、テロメラーゼ高活性の転移巣と低活性の原発巣のRNAを用いてマイクロアレイで解析したところ、いくつかの、発現パターンが異なる遺伝子が検出され、これらがテロメラーゼ活性化に関わっている可能性が示唆された。

Research Products
(12 results)

All Other

All Publications (12 results)

[Publications] Hiyama E., et al.: "Immunohistocheical detection of telomerase (hTERT) in human cancer tissues and a subset of cells in normal tissues."Neoplasia. (in press). (2001)
[Publications] Kumagai K. et al.: "Allelotype frequency of the thiopurine methyltransferase (TPMT) gene in Japanese"Pharmacogenetics. (in press). (2001)
[Publications] Hiyama E., et al.: "Biological characteristics of neuroblastoma with partial deletion in the short arm of chromosome 1,"Medical and Pediastric Oncology. 36・1. 67-71 (2001)
[Publications] Murakami I. et al: "p53 gene mutations are associated with shortened survival in patients with advanced non-small cell lung cancer : an analysis of medically managed patients"Clinical Cancer Research. 6・2. 526-301 (2000)
[Publications] Yoshikawa M. et al: "Microsomal epoxide hydrolase genotypes and chronic obstructive pulmonary disease in Japanese"Int J Mol Med. 5・1. 49-53 (2000)
[Publications] Ishioka S. et al: "Multiple Logistic Regression Analysis of Risk Factors for Development of Steroid-dependent Asthma in Elderly : A comparison with yougner asthmatics"Respiration. 68・1. 35-40 (2000)
[Publications] Hiyama E., et al.: "Multifocal Neuroblastoma : Biological Features and Surgical Aspects"Cancer. 88・8. 1955-1963 (2000)
[Publications] Tamagawa K: "Inhibitory effects of a lecithinized superoxide dismutase on bleomycin-induced pulmonary fibrosis in mice"Am J Respir Crit Care Med. 161・4. 1279-84 (2000)
[Publications] Mendoza C., et al.: "Allelotype and loss of heterozygosity around the L-myc gene locus in primary lung cancers"Lung cancer. 28・1. 117-125 (2000)
[Publications] Hiyama E., et al.: "Telomerase activity as a marker of breast carcinoma in fine-needle aspirated samples"Cancer. 90・4. 235-238 (2000)
[Publications] Ishioka S., et al.: "Effects of pranlukast, a cysteinyl leukotriene antagonist, on bronchial responsiveness to methacholine in aspirin-intolerant asthmatics treated with corticosteroids"Hiroshima J Med Sci. 49・2. 105-108 (2000)
[Publications] Ishioka S., et al.: "Clinical evaluation of idiopathic interstitial pneumonia and interstitial pneumonia associated with collagen vascular disease using logistic regression analysis."Intern Med. 39・3. 213-219 (2000)

2000 Fiscal Year Annual Research Report

肺癌細胞のテロメラーゼ活性・遺伝子変異と化学療法反応性の検討

Principal Investigator

檜山 桂子 広島大学, 医学部, 助手 (60253069)

Research Products

[Publications] Hiyama E., et al.: "Immunohistocheical detection of telomerase (hTERT) in human cancer tissues and a subset of cells in normal tissues."Neoplasia. (in press). (2001)

[Publications] Kumagai K. et al.: "Allelotype frequency of the thiopurine methyltransferase (TPMT) gene in Japanese"Pharmacogenetics. (in press). (2001)

[Publications] Hiyama E., et al.: "Biological characteristics of neuroblastoma with partial deletion in the short arm of chromosome 1,"Medical and Pediastric Oncology. 36・1. 67-71 (2001)

[Publications] Murakami I. et al: "p53 gene mutations are associated with shortened survival in patients with advanced non-small cell lung cancer : an analysis of medically managed patients"Clinical Cancer Research. 6・2. 526-301 (2000)

[Publications] Yoshikawa M. et al: "Microsomal epoxide hydrolase genotypes and chronic obstructive pulmonary disease in Japanese"Int J Mol Med. 5・1. 49-53 (2000)

[Publications] Ishioka S. et al: "Multiple Logistic Regression Analysis of Risk Factors for Development of Steroid-dependent Asthma in Elderly : A comparison with yougner asthmatics"Respiration. 68・1. 35-40 (2000)

[Publications] Hiyama E., et al.: "Multifocal Neuroblastoma : Biological Features and Surgical Aspects"Cancer. 88・8. 1955-1963 (2000)

[Publications] Tamagawa K: "Inhibitory effects of a lecithinized superoxide dismutase on bleomycin-induced pulmonary fibrosis in mice"Am J Respir Crit Care Med. 161・4. 1279-84 (2000)

[Publications] Mendoza C., et al.: "Allelotype and loss of heterozygosity around the L-myc gene locus in primary lung cancers"Lung cancer. 28・1. 117-125 (2000)

[Publications] Hiyama E., et al.: "Telomerase activity as a marker of breast carcinoma in fine-needle aspirated samples"Cancer. 90・4. 235-238 (2000)

[Publications] Ishioka S., et al.: "Effects of pranlukast, a cysteinyl leukotriene antagonist, on bronchial responsiveness to methacholine in aspirin-intolerant asthmatics treated with corticosteroids"Hiroshima J Med Sci. 49・2. 105-108 (2000)

[Publications] Ishioka S., et al.: "Clinical evaluation of idiopathic interstitial pneumonia and interstitial pneumonia associated with collagen vascular disease using logistic regression analysis."Intern Med. 39・3. 213-219 (2000)

檜山桂子広島大学, 医学部, 助手 (60253069)