实时定量PCR分析脂肪酸合酶在口腔鳞癌中的表达

Analysis of Gene Expression of Fatty Acid Synthase in Squamous Cell Carcinoma of the Oral Cavity by the Method of Real-time Quantitative PCR

目的利用实时定量PCR(RT-PCR)相对定量脂肪酸合酶(FAS)在口腔鳞癌、癌周以及正常口腔组织中的表达情况。方法切取口腔鳞癌患者新鲜手术标本的癌组织、癌周组织及正常组织,将组织切剪成碎块,提取细胞总RNA,经寡聚脱氧胸腺嘧啶逆转录,实时定量PCR扩增,针对内参照GAPDH基因相对定量FAS的表达,比较FAS在3种组织中的表达。结果FAS在癌组织中高表达,19例患者癌组织中的FASmRNA表达水平皆高于正常组织,最高达16倍(P<0.001)。13例癌旁组织表达水平较正常组织高(P<0.001)。结论口腔鳞癌组织和正常组织中的FAS表达有显著差异。实时定量PCR方法简便、快速、高效,并能相对定量内源性脂肪酸合酶在口腔鳞癌组织中的表达。

Objective To relatively quantify the gene expression of fatty acid synthase in squamous cell carcinoma, adjacent tissue, and some normal oral tissues by real-time quantitative PCR. Methods The tissues were collected fresh from surgical specimens. The collected tissues were minced. Then the total RNA was extracted. The RNA was reversely transcripted into cDNA with random prime. And then the cDNA was amplified by real-time quantitative PCR to quantify the gene expression of FAS according to an internal control GAPDH. The difference of FAS gene expression was compared between squamous cell carcinoma, adjacent tissue, and some normal oral tissues. Results The expression of FAS of squamous cell carcinoma was notably higher than the other two （P〈0.001）. Conclusion Real-time quantitative PCR provides a method for monitoring the expression of fatty acid synthetic activity in squamous cell carcinoma, adjacent and normal tissues.