Mutation Analysis of Heat Shock Cognate Protein 70 Gene in Colorectal Cancer

大腸癌熱誘導同質因子70之突變分析

10.6440/TZUCMJ.200212.0347

高瑞和(Ruey-Ho Kao)；郭明勳(Ming-Hsun Kuo)

熱誘導同質因子70 ； 大腸癌 ； 多形性變異 ； Heat shock cognate protein 70 ； colorectal cancer ； polymorphism

慈濟醫學雜誌

14卷6期（2002 / 12 / 01）

347 - 351

英文

目的：檢視大腸癌之熱誘導同質因子70(HSC70)之基因突變。材料與方法：選擇十個大腸癌的冷凍組織及十位健康人之周邊血液有核血球細胞，萃取其氧核糖核酸(DNA)。利用四對引子來放大HSC70之exon。放大的產物跑在膠片上，在紫外光檢視下將產物帶切下，並純化其中之DNA。利用自動定序儀器來定序這些DNA，所得結果與基因庫內HSC70 DNA 順序比對。結果：有十個順序變異被發現，其中有四個是已知的變界，六個是新的發現。有九個變異在大腸癌及正常人血液細胞中都有出現，表示其為同質多形性變異的表現。另一個變異只發生在兩位大腸癌，其位置在第3252基，第六個exon上。但因位於密碼的第三個位置，所以並不改變蛋白質的結構。結論：HSC70在大腸直腸癌並沒有有意義的基因改變。有六個同質多形性變異只發生在本實驗，而不發生在西方之研究，顯示東方人與西方人在基因體上的差異。

Objective: The aim of the study is to investigate possible mutations in heat shock cognate 70 gene (HSC70) in colorectal cancer. Materials and Methods: DNA was extracted from frozen tissue sections of 10 colorectal cancers and peripheral blood cells of 10 healthy people. The exons of HSC70 were PCR-amplified with 4 pairs of primers. The PCR products were run along a 1.5% agarose gel containing ethidium bromide and visualized under UV illumination. The PCR products were purified from the gel and directsequenced. The sequences were analysed according to GenBank access number Y00371. Results: Ten genetic alterations were found, and 6 were novel findings. Nine of them occurred both in colorectal cancer and healthy subjects, indicating polymorphisms. The remaining one, D6, found only in two colorectal cancers (20%), occurred at sequence 3252 in exon 6, but this sequence change did not alter the reading frame of HSC70 because of the third position wobble in codon 423. Conclusions: In the present study no significant genetic alterations were found in HSC70 for colorectal cancer; however several novel polymorphisms found only in this study demonstrated genomic differences between Asian and Western populations.