Heme oxygenase-1 和 NAD(P)H：quinone oxidoreductase 1 基因多形 性及砷甲基化代謝能力與泌尿道上皮癌的相關性研究
The Relationship among Heme oxygenase-1 and NAD(P)H：quinone oxidoreductase 1 Genetic Polymorphisms、Arsenic Methylation Capability and Urothelial Carcinoma
中文摘要
本研究為探討砷甲基化代謝能力、抗氧化酵素heme oxygenase-1(HO-1)和香菸代 謝相關酵素NAD(P)H:quinone oxidoreductase 1(NQO1)基因型在泌尿道上皮癌可
能扮演的角色，並評估砷、抽菸和HO-1 及 NQO1 基因型的交互作用。本研究自
民國91 年 9 月至 93 年 5 月間，於臺大醫院泌尿科門診所收泌尿道上皮癌病人 143 位。對照組為臺大醫院泌尿科門診尿路結石、前列腺肥大病人，臺北醫學大 學附設醫院老人健檢及臺北市立萬芳醫院成人健檢民眾，排除泌尿道上皮癌者。
經年齡性別配對之對照組共收案143 位。以標準化問卷訪視研究對象，收集人口
學基本資料、抽菸與飲酒習慣、職業暴露及個人與家族疾病史。取得研究對象同
意書後收集其血液尿液檢體。萃取血液DNA，利用聚合酵素連鎖反應
(Polymorphism chain reaction; PCR)增幅所需片段。用短縱列重複序列(Short Tandem Repeat Polymorphism, STRP)鑑定法，分析 HO-1 (GT)n 重複序列次數。
利用限制片段長度多形性(Restriction fragment length polymorphism)方法，測量 NQO1 C609T 基因多形性。利用高效能液相層析儀串聯氫化式原子吸收光譜儀測 定尿液三價砷、五價砷、單甲基砷酸與雙甲基砷酸濃度。結果發現，抽菸與農藥 為泌尿道上皮癌重要危險因子，隨著抽菸年數及累積抽菸量的增加泌尿道上皮 癌危險性亦隨之增加，顯示抽菸與泌尿道上皮癌有劑量效應關係。疾病組單甲基 砷酸百分比顯著高於對照組，雙甲基砷酸百分比與二級甲基化指標顯著低於對 照組，顯示疾病組砷甲基化代謝能力比對照組差。HO-1 (GT)n 重複序列基因多
形性及NQO1 基因多形性與泌尿道上皮癌無關。在調整年齡、性別與 HO-1 及
NQO1 基因型時，研究對象之砷甲基化代謝能力較差且抽菸及暴露農藥，泌尿 道上皮癌危險性是未暴露任何一個危險因子者之6.24 倍(95%信賴區間 1.84 — 21.22)。
英文摘要
The study to examine the potential role of arsenic methylation capability, antioxidant enzyme (heme oxygenase-1, HO-1) and cigarette smoking-related metabolic enzyme (NADPH:quinone oxidoreductase 1, NQO1) in urothelial carcinoma (UC), and to evaluate the interaction of arsenic methylation capability、cigarette smoking、HO-1 and NQO1 genotypes. Urothelial carcinoma 143 cases were recruited from the Department of Urology in National Taiwan University Hospital from September 2002 to May 2004. Age-sex matched 143 control subjects excluded urothelial carcinoma
were collected from Department of Urology NTUH, senile health examination from Taipei Medical University Hospital and adult health examination from Taipei Municipal WanFang Hospital. Interviewers who were well trained on interview techniques and carried out the standardized personal interviews based on structural questionnaire. Information obtained included the demographic characteristics, cigarette smoking and alcohol drinking habits, occupational exposure history, personal and family disease history. The study subjects who gave their consent were recruited for blood and urine samples. DNA was extracted from buffy coat, then performed by polymorphism chain reaction (PCR) to amplify special fragments.
Utilizing short tandem repeat polymorphism to analyze the number of HO-1 (GT)n repeats. Restriction fragment length polymorphism (RFLP) was used to determine NQO1 609 C to T polymorphism. Urine sample of subjects were measured by high performance liquid chromatography to separate arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA), and then quantified by hydride generator combined with atomic absorption spectrometry.
Cigarette smoking and pesticides exposure are important risk factors for UC, and the risk was increased with cigarette smoking duration and cumulative cigarette smoking exposure, with dose-response relationship. UC cases had a significantly higher percentage of MMA in urine than controls. The HO-1 (GT)n repeats polymorphism and NQO1 genotype are not associated with UC. After adjusted for age, sex, HO-1 genotype and NQO1 genotype, the study subjects with worse arsenic methylation capability, cigarette smoking and pesticide exposure, the odds ratio of UC were 6.24 (95% confidence interval 1.84 - 21.22) compared with those did not expose any risk factor.
