应用SYBR实时荧光定量Real—TimePCR法检测人宫颈癌MCPH1 mRNA表达

目的 应用SYBR实时荧光定量RT-PCR法检测MCPH1/BRIT1 mRNA的表达.方法 提取人宫颈癌总RNA,经逆转录PCR获得靶基因（MCPH1）及管家基因（GAPDH）的CDNA,采用SYBR Green 荧光实时定量法检测,以GAPDH基因作为内参,计算各组MCPH1 mRNA的相对表达量.结果 在31例宫颈癌标本中,MCPH1基因mRNA的表达,19例癌比正常低,1 例癌比正常高,其余标本无统计学意义;6例癌比癌旁低,1例癌比癌旁高,其余无统计学意义.结论 利用SYBR实时荧光定量RT-PCR检测出人宫颈癌中MCPH1基因mRNA的表达下调,为进一步研究MCPH1在宫颈癌中的作用及功能奠定了基础.

Detection of MCPH1 mRNA Expression in Cervical Cancer by Using SYBR Real-time Fluorescence Quantitative RT-PCR Method

Objective To detect MCPH1 mRNA expression in cervical cancer by using SYBR real-time fluorescence quantitative RT-PCR method. Methods The PCR products of MCPH1 gene and GAPDH housekeeping gerle were obtained by using RT-PCR after the total RNA was extracted from human cervical cancer. The MCPH1 mRNA expression was detected by SYBR Green real-time fluorescence quantitative RT-PCR method. GAPDH gene was used as the reference gene to calculate the relative expression of MCPHI mRNA of each group. Results In 31 cases of cervical cancer specimens, the MCPH1 mRNA expression was lower in 19 cases of cancer and higher in one case of cancer than in normal controls, and the remaining samples had no significant change. The MCPH1 mRNA expression was higher in 6 cases of cancer and lower in one case of cancer than in the adja- cent tissues, and the remaining specimens had no significant change. Conclusion The MCPH1 mRNA expression using SYBR Green real time fluorescence quantitative RT-PCR method in human cervical cancer was lower, which may provide the foundation for further on the role and function of MCPH1 in cervical cancer.