实时荧光定量RT-PCR检测宫颈癌组织中HIC1基因表达的应用研究

摘要

Objective To explore the expression level of hypermethylated in cancer KHICDmRNA in cervical cancer tissues by Real-time fluorescence quantitative RT-PCR (FQ-PCR) and discuss its significance in clinical diagnosis of cervical cancer. Methods 20 cases of invasive carcinoma of cervix and 20 cases of preinvasive cervix tissues were from Changshu general hospitals, which were diagnosed as cervical cancer clinically. 20 cases of normal control were collected from normal cervix. Total RNA was extacted and FQ-PCR was used to examine the levels of HIC1-mRNA expression in each of groups. Significance between them was analyzed by F test and t test of statistics. Results Relative quantitation of expression level of HIC1- mRNA (2-△△CT) in cervical cancer,preinvasive cervix tissues,and normal cervix tissues was 1. 0,0. 45 and 0. 15 differently. The significant difference between them was demonstrated by F test of statistics (F=4. 16,P<0. 05). By t test of statistics, HICl-mRNA expression level in cervical cancer tissues was less than that in preinvasive cervix tissues (t= 1. 74, P <0. 05),and lower than normal cervix tissues (t=2. 14,P<0. 01). HICl-mRNA expression level in preinvasive cervix tissues was lower than in normal cervix tissues( t=3. 69, P<0. 05). Conclusion Detection of the HICl-mRNA expression in cervical cancer tissues by real-time fluorescent relative quantitative RT-PCR (FQ-PCR) has a high sensitivity and specificity. It is reliable and easily performed. There is sigificantdifference in different cervical cancer staging. Low expression level of HICl-mRNA is related to metastasis of cervical cancer.%目的 实时荧光定量RT-PCR(FQ-PCR)技术检测宫颈癌组织中基因肿瘤超甲基化基因1(HIC1)mRNA表达水平,并探讨其表达差异在宫颈癌临床诊断治疗中的意义.方法 2010～2012年常熟地区综合医院临床妇科专业确诊并行手术治疗的宫颈癌疾病组织,按病理学分宫颈癌、宫颈原位癌两组,每组20例;收集20例正常宫颈组织设为对照组.提取其总RNA,FQ-PCR相对定量法检测每组中HIC1-mRNA的表达水平.统计学方差分析F检验,t检验分析三组间HIC1 mRNA表达量是否存在显著性差异.结果宫颈癌、宫颈原位癌及正常宫颈组织中HIC1 mRNA相对定量结果(2-△△CT)分别为1.0,0.45和0.15.F检验三组间表达量均呈现统计学显著性差异(F＝4.16,P＜0.05),t检验显示HIC1-mRNA在宫颈癌组织中的表达水平低于宫颈原位癌组织(t＝1.74,P＜0.05);宫颈原位癌中HIC1-mRNA表达水平低于正常宫颈组织(t＝2.14,P＜0.05);而宫颈癌组织中表达水平极显著低于正常组织(t＝3.69,P＜0.01).结论 FQ-PCR相对定量法可以敏感且特异地检测出HIC1-mRNA的表达水平,方法准确可靠,操作简便.宫颈癌不同分期中HIC1 mRNA表达水平存在统计学差异,HIC1基因的低水平表达与宫颈癌的侵袭转移有关.