Elucidation of the mechanism of cancer cell metastasis and permeation using active vitamin D as a molecular base and development of therapeutic agents

以活性维生素D为分子基础阐明癌细胞转移和渗透的机制并开发治疗剂

• 批准号: 15590083
• 负责人: OKANO Toshio
• 金额: $ 2.18万
• 依托单位: KOBE PHARMACEUTICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590083/
• 关键词: ACTIVE VITAMIN D ANALOGS; VITAMIN D RECEPTOR; CHEMICAL STRUCTURE-ACTIVITY RELATIONSHIP; VITAMIN D METABOLISM; APOPTOSIS; CELL GROWTH AND DIFFERENTIATION; TUMOR INVASION AND METASTASIS; ANGIOGENESIS; 腫瘍形成; 細胞増殖; がん転移・浸潤; ビタミンD代謝; 骨転移; 転移・浸潤

In the present study, we demonstrated from the in vitro and in vivo studies that 1α,25-dihydroxyvitamin D_3 (1α,25-D_3) and its derivatives are effective for the prevention and treatment of cancer. The results were follows. (1) It was found that 22-oxa-calcitriol, a low calcemic vitamin D analogue used clinically for the treatment of psoriasis and secondary hyperparathyroidism, is inactivated via through the pathway of either side-chain dehydration or isomerization of a hydroxy group at C-3 of the A-ring. In addition, it was found that the putative enzymes involved in the above pathways regulate the metabolism of la,25-D_3 cooperatively. (2) We confirmed that due to the lack of appropriate explanation for species difference (from animals to humans) of our results, employments of known-vitamin D metabolic enzymes expression systems were useful for the screening of the biological activity of vitamin D compounds. (3) We clarified that la,25-D_3 inhibits the growth of LLC-GFP cells by thro … More ugh the induction of G1 arrest, and inhibits the metastasis of cancer cells via the inhibition of MMP-2 and -9 expression, and suppress the angiogenesis of cancer cells via the inhibition of the expression of angiogenesis-inducing factors. (4) We clarified that using VDR gene targeting mice as a an animal model for the evaluation of cancer cell growth, l a,25-D_3 suppressed the tumor growth of LLC-GFP cells, which are VDR-positive, and the anti-tumorigenesis of l a,25-D_3 did not require the involvement of its calcemic action, and at both supraphysiological and normal serum concentrations, 1α,25-D_3 effectively suppressed the tumor growth. (5) I was found that 22-oxa-calcitriol suppressed the LLC-GFP cell growth without inducing any calcemic symptoms in animals. By the present study, we established the distinguished experimental model system for the screening of the anti-cancer activity of active vitamin D at gene, cellular and animal levels. Using these methods, it can be expected that the inter-cooperation between biology group and chemistry group will creates novel vitamin D analogues, especially having superior anti-cancer activity.In the present study, we demonstrated from the in vitro and in vivo studies that 1α,25-dihydroxyvitamin D_3 (1α,25-D_3) and its derivatives are effective for the prevention and treatment of cancer. The results were follows. (1)It was found that 22-oxa-calcitriol, a low calcemic vitamin D analogue used clinically for the treatment of psoriasis and secondary hyperparathyroidism, is inactivated via through the pathway of either side-chain dehydration or isomerization of a hydroxy group at C-3 of the A-ring. In addition, it was found that the putative enzymes involved in the above pathways regulate the metabolism of 1α,25-D_3 cooperatively. (2)We confirmed that due to the lack of appropriate explanation for species difference (from animals to humans) of our results, employments of known-vitamin D metabolic enzymes expression systems were useful for the screening of the biological activity of vitamin D compounds. (3)We clarified that 1α,25-D_3 inhibits the growth of LLC-GFP cells by through the induction of G1 arrest, and inhibits the metastasis of cancer cells via the inhibition of MMP-2 and -9 expression, and suppress the angiogenesis of cancer cells via the inhibition of the expression of angiogenesis-inducing factors. (4)We clarified that using VDR gene targeting mice as a an animal model for the evaluation of cancer cell growth, 1α,25-D_3 suppressed the tumor growth of LLC-GFP cells, which are VDR-positive, and the anti-tumorigenesis of 1α,25-D_3 did not require the involvement of its calcemic action, and at both supraphysiological and normal serum concentrations, 1α,25-D_3 effectively suppressed the tumor growth. (5)I was found that 22-oxa-calcitriol suppressed the LLC-GFP cell growth without inducing any calcemic symptoms in animals. By the present study, we established the distinguished experimental model system for the screening of the anti-cancer activity of active vitamin D at gene, cellular and animal levels. Using these methods, it can be expected that the inter-cooperation between biology group and chemistry group will creates novel vitamin D analogues, especially having superior anti-cancer activity. Less

本研究从体内外研究证实1α，25-二羟维生素D_3（1α，25-D_3）及其衍生物对肿瘤的预防和治疗有效。结果如下。(1)发现22-氧-骨化三醇，一种临床上用于治疗银屑病和继发性甲状旁腺功能亢进的低钙维生素D类似物，通过侧链脱水或A环C-3处羟基的异构化途径失活。此外，还发现参与上述途径的酶协同调节1a，25-D_3的代谢。(2)我们证实，由于缺乏适当的解释，我们的结果的物种差异（从动物到人类），采用已知的维生素D代谢酶表达系统是有用的筛选维生素D化合物的生物活性。(3)本实验表明，1a，25-D_3通过抑制LLC-GFP细胞的生长， ...更多信息 通过抑制MMP-2和MMP-9的表达来抑制癌细胞的转移，通过抑制血管生成诱导因子的表达来抑制癌细胞的血管生成。(4)我们阐明，使用VDR基因靶向小鼠作为评估癌细胞生长的动物模型，1 α，25-D_3抑制VDR阳性的LLC-GFP细胞的肿瘤生长，并且1 α，25-D_3的抗肿瘤作用不需要其钙调作用的参与，并且在超生理和正常血清浓度下，1α，25-D_3能有效抑制肿瘤生长。(5)发现22-氧杂-骨化三醇抑制LLC-GFP细胞生长，而不诱导动物中的任何钙化症状。本研究建立了从基因、细胞和动物水平筛选活性维生素D抗癌活性的优良实验模型体系。本研究通过体外和体内研究，证实了1α，25-二羟基维生素D_3（1α，25-D_3）及其衍生物对癌症的预防和治疗作用。结果如下。(1)It发现临床上用于治疗银屑病和继发性甲状旁腺功能亢进的低钙维生素D类似物22-氧-骨化三醇通过侧链脱水或A环C-3处羟基的异构化途径失活。此外，还发现上述途径中可能参与的酶协同调节1α，25-D_3的代谢。(2)We证实，由于我们的结果缺乏对物种差异（从动物到人类）的适当解释，使用已知的维生素D代谢酶表达系统对于筛选维生素D化合物的生物活性是有用的。(3)We阐明1α，25-D_3通过诱导G1期阻滞抑制LLC-GFP细胞生长，通过抑制MMP-2和MMP-9表达抑制癌细胞转移，通过抑制血管生成诱导因子表达抑制癌细胞血管生成。(4)We阐明了1α，25-D_3对VDR阳性的LLC-GFP细胞的肿瘤生长有抑制作用; 1 α，25-D_3的抗肿瘤作用不需要钙调作用; 1 α，25-D_3在超生理浓度和正常血清浓度下均能有效抑制肿瘤生长。(5)I发现22-氧杂-骨化三醇抑制LLC-GFP细胞生长，而不诱导动物中的任何钙化症状。本研究建立了从基因、细胞和动物水平筛选活性维生素D抗癌活性的优良实验模型体系。利用这些方法，可以预期生物组和化学组之间的相互合作将产生新的维生素D类似物，特别是具有上级抗癌活性的类似物。少

项目成果

Method for the determination of 25-hydroxyvitamin D in human plasma using high-performance liquid chromatography-tandem mass spectrometry

高效液相色谱-串联质谱法测定人血浆中25-羟基维生素D的方法

• 发表时间: 2005
• 期刊: Anal.Chem. 77(9)
• 作者: Nakagawa K.;Okano T.;et al.;N.Tsugawa
• 通讯作者: N.Tsugawa

Long-term hospitalization during pregnancy is a risk factor for vitamin D deficiency in neonates

孕期长期住院是新生儿维生素D缺乏的危险因素

• 发表时间: 2003
• 期刊: J. Bone Miner. Metab. 21
• 作者: 鎌尾 まや;岡野登志夫;T.Sakaki;Y.Mizushina;M.Kamao;K.Nishimura
• 通讯作者: K.Nishimura

Selective Inhibition of Mammalian DNA Polymerase γ by Vitamin D_2 and D_3

维生素 D_2 和 D_3 对哺乳动物 DNA 聚合酶 γ 的选择性抑制

• 发表时间: 2003
• 期刊: J.Pharmacol.Sci. 92
• 作者: Okano T.;et al.
• 通讯作者: et al.

岡野 登志夫: "CLINICAL CALCIUM 13(7)「日本人のビタミンDとカルシウムの摂取状況」"医薬ジャーナル社. 42-51 (2003)

Toshio Okano：“CLINICAL CALCIUM 13(7)“日本人维生素 D 和钙的摄入状况””Iyaku Journal Inc. 42-51 (2003)

Regulation of gene expression of epithelial calcium channels in intestine and kidney of mice 1α,25-dihydroxyvitamin D_3

小鼠肠肾上皮钙通道基因表达调控1α,25-二羟基维生素D_3

• 发表时间: 2004
• 期刊: J. Steroid Biochem. & Mol. Biol. 89-90
• 作者: Nakagawa K.;Okano T.;et al.;N.Tsugawa;Y.Suhara;M.Kamao;K.Nakagawa;K.Nakagawa;K.Nakagawa;N.Sawada;M.Kamao;T.Okano
• 通讯作者: T.Okano