重庆医学Issue(18)：2323-2325,3.DOI:10.3969/j.issn.1671-8348.2014.18.021
SYBR实时荧光定量PCR检测人乳腺癌细胞AnnexinⅡmRNA方法的建立及应用
Establishment and application of SYBR real time fluorescence quantitative PCR for detecting Annexin Ⅱ mRNA of human breast cancer cells
摘要
Abstract
Objective To establish a SYBR Green based real-time fluorescence quantitative PCR method for detecting human Annexin Ⅱ mRNA expression,and to detect the level of Annexin Ⅱ mRNA in human breast cancer cells MCF-7 and MDA-MB-231.Methods The specific primers were designed according to the conserved sequence of human Annexin Ⅱ gene.Total RNAs were extracted from human breast cancer cells(MCF-7,MDA-MB-231),then RNAs were transcribed reversely into cDNAs.The plasmid standards were constructed.The relative expression levels of human Annexin Ⅱ mRNA in human breast cancer cells were detected by this method.Results The square(r2 )of correlation coefficient of the standard curve in this method was 0.997,the melting curve analysis showed the single peak.The the intra-batch and inter-batch variable coefficients in the pGM-T Annexin Ⅱplasmid standard substance were 6.2%,7.8% and 9.1%,12.3% respectively.The further study indicated that AnnexinⅡ mRNA in MDA-MB-231 was higher than that in MCF-7(P<0.01).Conclusion The established SYBR Green real time fluorescence quan-titative PCR method for detecting human AnnexinⅡ is of good specificity and repeatability and can be used for quantitatively detec-ting AnnexinⅡ mRNA in breast cancer cells.关键词
乳腺肿瘤/AnnexinⅡ/SYBR Green实时荧光定量PCRKey words
breast-neoplasms/AnnexinⅡ/SYBR-Green-real-time-fluorescence-quantitative-PCR引用本文复制引用
郭变琴,黄学梅,吴立翔..SYBR实时荧光定量PCR检测人乳腺癌细胞AnnexinⅡmRNA方法的建立及应用[J].重庆医学,2014,(18):2323-2325,3.基金项目
重庆市自然科学基金资助项目（2006BB5427）。 （）
