2001 年度実績報告書

抗酸化剤によるヒト口腔扁平上皮癌細胞死におけるHIF-1αの挙動

研究課題

研究課題/領域番号	11671853
研究機関	明海大学
研究代表者	坂上宏明海大学, 歯学部, 教授 (50138484)
研究分担者	柏俣正典朝日大学, 歯学部, 教授 (30152630)
キーワード	抗酸化剤 / ヒト口腔扁平上皮癌 / 細胞死 / アポトーシス / HIF-1α / MnSOD / bax / NO
研究概要	ヒト口腔扁平上皮癌HSC-2細胞は、低酸素状態においては、HIF-1αのHREへの結合が増加する。この時、抗酸化剤のNACや細胞内移行型のGSH ethylesterは、この反応を抑制するが、アスコルビン酸の抑制効果は、極僅かなものであった。一方、プロモーター領域にヒトVEGFの、HREをtandemにrepeatしたluciferase遺伝子を安定に発現する変異株を作成して、レポーターアツセイで、HIF-1の活性化を検討したが、通常酸素状態で高濃度のアスコルビン酸を添加6時間後で、全く誘導が認められなかった。これに対して、HL-60細胞においては、高濃度(5mM)アスコルビン酸によるアポトーシス誘導の過程では、濃度依存的にHIF-1αのmRNAの発現が、添加1時間以内に約2.5倍ほど増加することがRT-PCR解析で判明した。これは、アスコルビン酸添加により培養液中の酸素が消費して、hypoxiaが誘導されること、そして酸素供給に必要なタンパク質を発現するように細胞側が対応することを示している。これらのタンパク質を同定すること、また、HIF-1αのmRNAの誘導にもかかわらずHIF-1の活性化に至らずに細胞死に至る機構の解明は、今後の研究テーマである。緑茶主成分のEGCGや、イソフラボンは、リポ多糖によるマウスマクロファージRaw264.7細胞の活性化(NO、シトルリン、アスパラギン、TNF産生)を抑制するとともに、Raw264.7細胞にアポトーシス(TUNEL陽性細胞の出現、baxの発現促進、iNOS, SOD1の発現抑制)の誘導した。Baxの発現は、(抗酸化作用を示す)低濃度では低下するが、(酸化作用を示す)高濃度では増加し、2相性の調節機構が示唆された。高濃度の抗酸化剤は、HIF-1αとは対照的に、ミトコンドリアのMnSOD活性やMnSOD mRNAの発現を有意に低下させた。アポトーシスの進行過程で、細胞が低酸素・酸素ストレスに対処する機構として、HIF-1αやMnSODmRNAの発現調節を独立的に働かせている可能牲を示唆する。

研究成果
(26件)

すべてその他

すべて文献書誌 (26件)

[文献書誌] Imai K: "Radical scavenging activity and cytotoxicity of phenethyl ether and cinnamic aminde derivatives"Anticancer Res. 22(印刷中). (2002)
[文献書誌] Miyamoto M: "Tumor-specificity and radical scavenging activity of poly-herbal formula"Anticacner Res. 22(印刷中). (2002)
[文献書誌] Zhang W: "Decline of superoxide dismutase activity during antioxidant-induced apoptosis in HL-60 cells"Anticacner Res. 22(印刷中). (2002)
[文献書誌] Suzuki F: "Stimulation of arginine consumption and asparagine production in LPS-activated macrophages"Anticacner Res. 22(印刷中). (2002)
[文献書誌] Tashiro M: "Effects of prenylfiavones from Sophora species on growth and activation of mouse macrophage-like cell lines"Anticacner Res. 22(印刷中). (2002)
[文献書誌] Taniguchi S: "Production of bioactive triterpenes by Eriobotyra Japonica calli"Phytochemistry. 59. 315-323 (2002)
[文献書誌] Watanabe K: "Cycloartane glycosides from the Rhizomes of Cimjicifuga racemosa and their cytotoxic activities"Chem Pharm Bull. 50. 121-125 (2002)
[文献書誌] Motohashi N: "Biological activity of a vegetable,"Anastasia red", a series of sweet pepper"Phytotherapy Res. (印刷中). (2001)
[文献書誌] Zhang W: "Synergistic cytotoxic action of vitamin C and vitamin K_3"Anticancer Res. 21. 3439-3444 (2001)
[文献書誌] Motohashi N: "Biological activity of a fruit vegetable, "Anastasia green", a series of sweet pepper"In vivo. 15. 437-442 (2001)
[文献書誌] Shirataki Y: "Antioxidative activity of a folklore plant Allium vicorialis L"Anticancer Res. 21. 3331-3340 (2001)
[文献書誌] Jiang Y: "Inhibition of chlorogenic acid-induced cytotoxicity by CoCl_2"Anticancer Res. 21. 3349-3354 (2001)
[文献書誌] Shi Y-Q: "Cytotoxic and DNA damage-inducing activities of low molecular weight phenols from rhubarb"Anticancer Res. 21. 2847-2854 (2001)
[文献書誌] Shirataki Y: "Relationship between cytotoxic activity and radical intensity of isoflavones from Sophora Species"Anticancer Res. 21. 2643-2648 (2001)
[文献書誌] Sakagami H: "Production of hydrogen peroxide and methionine sulfoxide by epigallocatechin gallate and antioxidants"Anticancer Res. 21. 2633-2642 (2001)
[文献書誌] Sakagami H: "Changes in amino acid pool and utilization during the apoptosis of HL-60 cells induced by epigallocatechin gallate or gallic acid"Anticancer Res. 21. 2441-2448 (2001)
[文献書誌] Okayasu H: "Cytotoxic activity of vitamin K_1, K_2 and K_3 against human oral tumor cell lines"Anticancer Res. 21. 2387-2392 (2001)
[文献書誌] Aoki K: "Modification of apoptosis-inducing activity of gallic acid by saliva"Anticancer Res. 21. 1879-1883 (2001)
[文献書誌] 坂上宏: "基礎研究と口腔がん治療への応用の可能性について(その3)ポリフェノールとがん治療"Hosp. Dent.. 13. 2-13 (2001)
[文献書誌] Kuroda M: "Steroidal glycosides from the bulbs of Camsssia leichtlinii and their cytotoxic activities"Chem Pharm Bull. 492. 726-731 (2001)
[文献書誌] Furuya S: "Cytotoxic activity of saponins from Camassia leichtlinii against human oral tumor cell lines"Anticancer Res. 21. 959-964 (2001)
[文献書誌] Terasawa K: "Cytotoxic activity of 5-benzoylimidazole and related compounds against human oral tumor cell lines"27. 1081-1086 (2001)
[文献書誌] Shirataki Y: "In vitro biological activity of prenylflavanone"Anticancer Res. 21. 275-280 (2001)
[文献書誌] Asano K: "Tumor-specific action of sodium 5, 6-benzylidene-L-ascorbate in N-nitrosodiethylamine-administered mouse model"Anticancer Res. 21. 281-284 (2001)
[文献書誌] Shi Y-Q: "Cytotoxic flavonoids with isoprenoid groups from Morus mongolica"J Natural Products. 64. 181-188 (2001)
[文献書誌] Hou A-J: "Benzophenones and xanthones with isoprenoid groups from Cudrania cochinchinensis. J"J Natural Products. 64. 65-70 (2001)

2001 年度 実績報告書

抗酸化剤によるヒト口腔扁平上皮癌細胞死におけるHIF-1αの挙動

研究代表者

坂上 宏 明海大学, 歯学部, 教授 (50138484)

研究成果

[文献書誌] Imai K: "Radical scavenging activity and cytotoxicity of phenethyl ether and cinnamic aminde derivatives"Anticancer Res. 22(印刷中). (2002)

[文献書誌] Miyamoto M: "Tumor-specificity and radical scavenging activity of poly-herbal formula"Anticacner Res. 22(印刷中). (2002)

[文献書誌] Zhang W: "Decline of superoxide dismutase activity during antioxidant-induced apoptosis in HL-60 cells"Anticacner Res. 22(印刷中). (2002)

[文献書誌] Suzuki F: "Stimulation of arginine consumption and asparagine production in LPS-activated macrophages"Anticacner Res. 22(印刷中). (2002)

[文献書誌] Tashiro M: "Effects of prenylfiavones from Sophora species on growth and activation of mouse macrophage-like cell lines"Anticacner Res. 22(印刷中). (2002)

[文献書誌] Taniguchi S: "Production of bioactive triterpenes by Eriobotyra Japonica calli"Phytochemistry. 59. 315-323 (2002)

[文献書誌] Watanabe K: "Cycloartane glycosides from the Rhizomes of Cimjicifuga racemosa and their cytotoxic activities"Chem Pharm Bull. 50. 121-125 (2002)

[文献書誌] Motohashi N: "Biological activity of a vegetable,"Anastasia red", a series of sweet pepper"Phytotherapy Res. (印刷中). (2001)

[文献書誌] Zhang W: "Synergistic cytotoxic action of vitamin C and vitamin K_3"Anticancer Res. 21. 3439-3444 (2001)

[文献書誌] Motohashi N: "Biological activity of a fruit vegetable, "Anastasia green", a series of sweet pepper"In vivo. 15. 437-442 (2001)

[文献書誌] Shirataki Y: "Antioxidative activity of a folklore plant Allium vicorialis L"Anticancer Res. 21. 3331-3340 (2001)

[文献書誌] Jiang Y: "Inhibition of chlorogenic acid-induced cytotoxicity by CoCl_2"Anticancer Res. 21. 3349-3354 (2001)

[文献書誌] Shi Y-Q: "Cytotoxic and DNA damage-inducing activities of low molecular weight phenols from rhubarb"Anticancer Res. 21. 2847-2854 (2001)

[文献書誌] Shirataki Y: "Relationship between cytotoxic activity and radical intensity of isoflavones from Sophora Species"Anticancer Res. 21. 2643-2648 (2001)

[文献書誌] Sakagami H: "Production of hydrogen peroxide and methionine sulfoxide by epigallocatechin gallate and antioxidants"Anticancer Res. 21. 2633-2642 (2001)

[文献書誌] Sakagami H: "Changes in amino acid pool and utilization during the apoptosis of HL-60 cells induced by epigallocatechin gallate or gallic acid"Anticancer Res. 21. 2441-2448 (2001)

[文献書誌] Okayasu H: "Cytotoxic activity of vitamin K_1, K_2 and K_3 against human oral tumor cell lines"Anticancer Res. 21. 2387-2392 (2001)

[文献書誌] Aoki K: "Modification of apoptosis-inducing activity of gallic acid by saliva"Anticancer Res. 21. 1879-1883 (2001)

[文献書誌] 坂上 宏: "基礎研究と口腔がん治療への応用の可能性について(その3)ポリフェノールとがん治療"Hosp. Dent.. 13. 2-13 (2001)

[文献書誌] Kuroda M: "Steroidal glycosides from the bulbs of Camsssia leichtlinii and their cytotoxic activities"Chem Pharm Bull. 492. 726-731 (2001)

[文献書誌] Furuya S: "Cytotoxic activity of saponins from Camassia leichtlinii against human oral tumor cell lines"Anticancer Res. 21. 959-964 (2001)

[文献書誌] Terasawa K: "Cytotoxic activity of 5-benzoylimidazole and related compounds against human oral tumor cell lines"27. 1081-1086 (2001)

[文献書誌] Shirataki Y: "In vitro biological activity of prenylflavanone"Anticancer Res. 21. 275-280 (2001)

[文献書誌] Asano K: "Tumor-specific action of sodium 5, 6-benzylidene-L-ascorbate in N-nitrosodiethylamine-administered mouse model"Anticancer Res. 21. 281-284 (2001)

[文献書誌] Shi Y-Q: "Cytotoxic flavonoids with isoprenoid groups from Morus mongolica"J Natural Products. 64. 181-188 (2001)

[文献書誌] Hou A-J: "Benzophenones and xanthones with isoprenoid groups from Cudrania cochinchinensis. J"J Natural Products. 64. 65-70 (2001)

2001 年度実績報告書

坂上宏明海大学, 歯学部, 教授 (50138484)

[文献書誌] 坂上宏: "基礎研究と口腔がん治療への応用の可能性について(その3)ポリフェノールとがん治療"Hosp. Dent.. 13. 2-13 (2001)