Identification of high risk group for prostate cancer by investigation of vitamin D receptor gene

维生素D受体基因检测识别前列腺癌高危人群

• 批准号: 12557133
• 负责人: HATANO Tadashi
• 金额: $ 2.5万
• 依托单位: University of the Ryukyus
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557133/
• 关键词: vitamin D receptor gene; urological disease; PCR-RFLP; single nucleotide polymorphism; SNP; microsatellite analysis; high risk group; 泌尿器科疾患; 前立腺癌; hight risk group

Firstly, single nucleotide polymorphisms (SNPs) of vitamin D receptor gene were examined using three PCR-RFLP markers spanning between intron 8 and exon 8 in the healthy volunteers and the patients with urological problems including prostate cancer, other cancers and urolithiasis. Three restriction sites were ApaI, BamHI and TaqI. Genotypes in 101 of control samples were, TT (74.3%), Tt (24.8%) and tt (1.0%) for TaqI, BB (4.0%), Bb (27.7%) and bb (68.3%) for BamHI, AA (11.9%), Aa (43.6%), and aa (44.6%) for ApaI. In the patient with urolithiasis, t allele was indicated as a risk factor for the recurrent disease. Secondly, deletion mapping of chromosome 6 in 40 cases of renal cell carcinoma was performed. Six microsatellite markers on 6q, 22.3 cM from D6S975 to D6S1577 were analyzed. 13 cases out of 40 (33%) were found to have LOH at least one out of 5 markers. Among the cases with LOH, LOH in the 17.1 cM region with D6S311 and/or the 9.8 cM region with D6S441 were frequently observed. No remarkable findings were seen in prostate cancer cases in the same analysis. Thirdly, mutation analysis of BRG1 gene using PCR-SSCP was performed. BRG1 gene is located at 19p where deletion is frequently seen in prostate cancer. Variation of the genotype of BRG1 gene was observed in exon 9. Direct sequencing revealed SNP33978 of T/T, T/C, C/C. The patients with T/T genotype of SNP33978 were diagnosed at significantly younger ages (p=0.03). Young patients (age<62 years) with T/T genotype were more frequently to have high grade, advanced stage and metastatic tumors than patients with the T/C or C/C genotype (p=0.0134). It was suggested to be useful in preventive medicine, in decision making of therapeutic options and in predicting prognosis patients with cancer to analyze the variation and the combination of several germline SNPs.

首先，采用PCR-RFLP方法检测了健康志愿者和前列腺癌、其他癌症、尿石症患者维生素D受体基因第8内含子和第8外显子之间的单核苷酸多态性（single nucleotide polymorphism，SNPs）。3个酶切位点分别为ApaI、BamHI和TaqI。101例对照样本的TaqI基因型为TT（74.3%）、Tt（24.8%）和tt（1.0%），BamHI基因型为BB（4.0%）、Bb（27.7%）和bb（68.3%），ApaI基因型为AA（11.9%）、Aa（43.6%）和aa（44.6%）。在尿石症患者中，t等位基因是尿石症复发的危险因素。其次，对40例肾细胞癌进行了6号染色体缺失定位。分析了从D 6S 975到D 6S 1577的6个微卫星标记，位于6 q，22.3cM。40例中有13例（33%）在5个标记物中至少有1个存在洛丢失。在洛缺失的病例中，D 6S 311位点的17.1 cM区域和/或D 6S 441位点的9.8 cM区域的洛最常见。在同一分析中，前列腺癌病例中未观察到显著结果。第三，采用PCR-SSCP技术对BRG 1基因进行突变分析。BRG 1基因位于19 p，在前列腺癌中常见缺失。BRG 1基因第9外显子存在基因型变异。直接测序显示T/T、T/C、C/C的SNP 33978。SNP 33978 T/T基因型患者的确诊年龄明显较低（p=0.03）。年轻患者（<62岁）T/T基因型较T/C或C/C基因型更易发生高级别、晚期和转移性肿瘤（p=0.0134）。分析生殖系SNPs的变异及其组合对癌症的预防、治疗方案的制定和预后的预测具有重要意义。

项目成果

Oya M., Ohtsubo M., Takayanagi A., Tachibana M., Shimizu N., Murai M.: "Constitutive activation of nuclear factor-kappaB prevents TRAIL-induced apoptosis in renal cancer cells"Oncogene. 20. 3888-3896 (2001)

Oya M.、Ohtsubo M.、Takayanagi A.、Tachibana M.、Shimizu N.、Murai M.：“核因子-kappaB 的组成型激活可防止肾癌细胞中 TRAIL 诱导的细胞凋亡”癌基因。

秦野 直: "腎穿刺・経皮的腎造影・腎瘻造設術"臨床泌尿器科. 56・4. 101-107 (2002)

秦野直树：“肾穿刺、经皮肾造影和肾造口术”《临床泌尿外科》56・4（2002）。

Nishiyama T., Tachibana M., Horiguchi Y., Nakamura K., Ikeda Y., Takesako K., Murai M.: "Immunotherapy of bladder cancer using autologous dendritic cells pulsed with human lymphocyte antigen-A24-specific MAGE-3 peptide"Clin Cancer Res. 7. (2001)

Nishiyama T.、Tachibana M.、Horiguchi Y.、Nakamura K.、Ikeda Y.、Takesako K.、Murai M.：“使用人淋巴细胞抗原 A24 特异性 MAGE-3 肽脉冲的自体树突状细胞进行膀胱癌的免疫治疗

Ogawa Y., Sugaya K, Koyama Y, Hatano T.: "Impact of citrate therapy on the circadian rhythm of urinary magnesium ammonium phosphate saturation I normal individuals"Int J Urol. 7. 287-291 (2000)

Okawa Y.、Sugaya K、Koyama Y、Hatano T.：“柠檬酸盐疗法对正常个体尿磷酸镁铵饱和度昼夜节律的影响”Int J Urol。

小川由英: "今日の診断指針第5版 泌尿器・男性性器疾患-腎・尿管結石"東京医学社. 1538-1541 (2002)

Yoshihide Okawa：“当今的泌尿和男性生殖器疾病诊断指南第 5 版 - 肾脏和输尿管结石”Tokyo Igakusha 1538-1541 (2002)。