以肌动蛋白为标准化参数在甲状腺乳头状癌中c-fos表达的研究

The study of oncogene c-fos expression in papillary thyroid carcinoma by use of β-actin as a normalization parameter

目的 :引入细胞管家基因 (β-actin)作为多重扩增的内参照物以达到资料比较的标准化 ,用于研究c -fos在甲状腺乳头状癌中的表达。方法 :14例甲状腺乳头状癌组织标本 ;21例正常对照包括3例乳头状癌病变旁正常甲状腺组织和18例腺瘤病变旁正常甲状腺组织。采用Trizol法提取总RNA ,逆转录 -聚合酶链反应法 (RT -PCR)获取cDNA ,多重聚合酶链反应 (MultiplePCR)法同时扩增371bp的c -fos和227bp的β-actin。扩增产物行1.5%琼脂糖凝胶电泳 ,用UVP凝胶成像分析系统的GelBase/GelBiot -Pro分析软件获取各标本c-fos和β-actin的密度值 ,计算两者的比值。结果 :正常对照和乳头状癌组c -fos/β-actin比值分别为 (x±s) :0.6845±0.0753和0.5938±0.0969 ,两组存在着显著差异(t=2.954,P<0.01) ;经自身配对t检验 ,3例患者的癌变组织明显低于与此癌病变旁的正常组织(t=9.205 ,P<0.05)。结论 :c -fos在甲状腺乳头状癌组织中的表达明显低于正常对照。

Objective: β-actin, one of housekeeping genes, was used in multiple PCR as the internal reference to normalize the data in the study of c-fos expression in thyroid papillary carcinoma.Methods:14 samples of fresh thyroid tumor tissue in vivo were obtained after thyroidectomy and proved to be papillary thyroid carcinoma. 21 samples of thyroid normal control tissues included 3 cases adjacent to its papillary thyroid carcinoma tissue and 18 cases adjacent to thyroid adenoma. Total RNA was extracted from thyroid samples by TRIzol method and then cDNA were obtained by RT-PCR.371 bp of c-fos and 227 bp of β-actin were amplified simultaneously in a multiple-PCR.After co-amplification products were separated on 1.5% agarose gel, the products was analyzed by GelBase/Biot-Pro software of UVP system. The ratio of c-fos to β-actin were calculated in order to normalize the data.Results:The ratio of c-fos to β-actin(x±s)in normal control group and papillary thyroid carcinoma group was 0.6845±0.0753 and 0.5938±0.0969 respectively. There was significant difference between two groups (t=2.954,P<0.01). The ratio in 3 cases of papillary thyroid carcinoma was significantly lower than that in normal control adjacent to them by self-pair t-test (t=9.205,P<0.05). Conclusion:The expression of c-fos in papillary thyroid carcinoma group was significantly lower than that in normal control group.