实时定量PCR分析ST13基因在肿瘤细胞株中的表达

Analysis of gene expression of ST13 in tumor cell lines by real-time quantitative PCR

目的 利用实时定量PCR(real timequantitativePCR)相对定量ST1 3基因在不同肿瘤细胞株的表达情况。方法 提取肿瘤细胞株SW 6 2 0、A5 4 9、Bxpc 3、SMMC 772 1、RKO和Bcap 37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ST1 3基因的表达 ,比较ST1 3基因在不同组织癌细胞株中的表达。结果 ST1 3基因在 6种肿瘤细胞株中表达高 ,相当于GAPDH基因表达的 1 %～ 2 0 % ,细胞株之间ST1 3基因表达量分高、中、低三类 (P <0 .0 5 ) ,呈现Bcap 37、RKO >SMMC 772 1和Bxpc 3>A5 4 9、SW6 2 0。 结论 本方法简便、快速、高效地反映了ST1 3基因在细胞株中的表达情况 。

Objective To quantify relatively the gene expression of ST13 in six cell lines by real time quantitative PCR.Methods The total RNA was extracted from cell lines SW620,A549,Bxpc 3,SMMC 7721,RKO and Bcap 37.The RNA was reversely transcripted into cDNA with oligo dT.Then the cDNA was amplified by real time quantitative PCR to quantify the gene expression of ST13 according to an internal control GAPDH.The difference of ST13 gene expression was compared between the cell lines.Results The gene expression of ST13 was as much high as 1% to 20% of GAPDH in the six cell lines respectively.The expression level was divided into three sorts (high,middle and low),showing Bcap 37,RKO>SMMC 7721 and Bxpc 3>A549,SW620.Conclusion Real time PCR provides a method for monitoring the gene expression of ST13 in cell lines.And it is a useful technique means on gene research.