Taqman定量PCR检测宫颈癌样本中CXCL12和CXCR4基因表达水平

摘要

Objective To investigate the effect of detection of mRNA expression levels of CXCL12 and CXCR4 in early diagnosis and lymphatic metastatic diagnosis of cervical cancer.Methods From January 2015 to September 2016,113 patients with cervical cancer treated at department of gynecology and obstetrics were taken as cancer group.Meanwhile,20 patients with healthy cervical tissue were taken as control group.The results of CXCL12 and CXCR4 in different pathological types,and the accuracy,specificity and repeatability of the detection of mRNA expression of CXCL12 and CXCR4 were observed.The changes of CXCL12 and CXCR4 mRNA expressions between the cervical cancer samples and normal cervical tissues as well as among the various clinicopathological parameters were compared.Results The accuracy,specificity and measuring linearity of Taqman real-time PCR met the design requirements.Standard curve results showed good repeatability.In cervical cancer tumor,the expression levels of CXCR4 mRNA were increased (2-△Ct =3.01 ± 0.09).However,there was no significant difference in CXCR4 mRNA expression among clinical stages and pathological patterns (P ＞ 0.05).When the tumor was metastatic,the expression levels of CXCL12 mRNA were reduced (2-△Ct =0.09 ± 0.02) that was significantly lower than that in cancer without metastasis (2-aCt =0.19 ± 0.20) (P ＜ 0.05).Conclusion CXCL12 and CXCR4 could be used as biomarkers that may play an important role in early diagnosis of cervical cancer.The real time PCR combined the calculation method of 2-△Ctcould reach idea results that will be suitable for clinical further promotion.%目的 探讨CXCL12和CXCR4基因检测在宫颈癌病变筛查中对宫颈癌早期诊断及其淋巴结转移检测的效果.方法 选择2015年1月至2016年9月在我院妇产科就诊的宫颈癌患者113例为癌症组,另选同期正常宫颈组织20例作为正常对照.观察不同病理分型中CXCL12和CXCR4检测结果,以及CXCL12和CXCR4 mRNA检测准确性、特异性以及重复性等指标,分析不同临床病理指标的宫颈癌样本和正常宫颈样本中CXCL12和CXCR4mRNA的表达量的变化.结果 多重荧光定量体系的准确性、特异性和检测线性结果均满足设计要求,采用标准曲线法评估试剂的重复性.结果显示,标准曲线呈现良好的线性梯度和重复性.对实际临床样本进行检测,确定宫颈癌组织中CXCR4mRNA表达量和正常组织中上调明显,2-△Ct等于或大于(3.01±0.09),不同临床分型结果或者肿瘤病理类型差异无统计学意义(P＞0.05);而当淋巴结组织中出现转移的情况下,CXCL12的下调至(0.09±0.02),与无淋巴组织转移的肿瘤(0.19±0.2)差异有统计学意义(P＜0.05).结论 CXCL12和CXCR4可以作为早期生物标志物,可以在宫颈癌早期检测中发挥重要作用,采用荧光定量PCR方法结合2-△Ct计算方法具有理想的效果,适合临床进一步研究和推广.