摘要
目的开发一种灵敏度、特异性、准确性的检测微卫星不稳定(MSI)状态的PCR扩增方法,以期为中国人群MSI检测位点提供最佳临床数据。方法选取郑州大学第一附属医院2017年至2019年收治的201例结直肠癌患者样本,利用PCR法检测5个标准MSI检测位点(BAT25,BAT26,D2S123,D5S346和D17S250)。本次研究对结直肠癌临床样本进行HE染色处理,样本坏死的情况通过显微镜进行观察和评估。利用免疫组化的方法检测错配修复(MMR)蛋白,分析MSH2、MSH6、MLH1和PMS2的表达缺失情况。卡方检验MSI与临床病理特征参数的关系。结果由本PCR方法检测确定本次研究的201例CRC患者样本的微卫星状态,其中5例(12.4%)为高频微卫星不稳定(MSI-H)型,176例(87.6%)属于低频微卫星不稳定(MSI-L)和微卫星稳定(MSS)型。本次研究结果显示CRC患者MSI状态与肿瘤发生年龄、肿瘤原发灶部位、TNM分期以及病理分型方面差异存在统计学意义(P<0.05),在患者性别、分化程度、KRAS、NRAS、BRAF基因变异方面差异无统计学意义(P>0.05)。对MSI状态的检测结果与MMR免疫组化的检测结果进行一致性分析,Kappa值为0.978,P值小于0.001。NCI Panel检测MSI的灵敏度为96.2%,特异性为100%;阳性预测值和阴性预测值分别为100%和99.4%。结论微卫星不稳定(MSI)状态与CRC在发病年龄、肿瘤原发灶部位、TNM分期以及病理分型方面的临床病理特征关系密切。通过检测NCI的5个标准检测位点,本PCR方法是一种具有优良灵敏度、特异性和准确性的MSI检测方法。
Objective To develop a sensitive,specific and accurate PCR method for detecting microsatellite instability(MSI)status,to provide the best clinical data for MSI detection sites in the Chinese population.Methods Totally 201 samples of colorectal cancer patients were collected.Five standard MSI loci(BAT25,BAT26,D2 S123,D5 S346 and D17 S250)were detected by PCR.The necrosis of colorectal cancer samples stained by Hematoxylin-eosin staining,and evaluated by microscope.The expression of mismatch repair(MMR)proteins MSH2,MSH6,MLH1,and PMS2 were detected by immunohistochemistry to analyze the expression loss of MMR protein.The association was analyzed between the MSI state and clinic pathological characteristics.Results Among 201 samples,25(12.4%)were microsatellite instability-high(MSI-H),176(87.6%)were microsatellite instability-low(MSI-L),and microsatellite stability(MSS).CRC patients with MSI-H and MSI-L&MSS had difference in age,tumor location,TNM and histological type(P<0.05),but had no obvious difference in gender,tumor differentiation and KRAS,NRAS,BRAF gene metastasis(P>0.05).The kappa-value of the NCI panel and MMR IHC was 0.978,The P-value was less than 0.001.The sensitivity and specificity of NCI Panel in detecting MSI were 96.2%,100%,and the positive predictive value and negative predictive value were 100%and 99.4%,respectively.Conclusion There is a close relationship between MSI state and the clinic pathological characteristics of CRC.This PCR method is sensitive,specific,accurate and anti-interference for detecting MSI status.
作者
李震
姚弘虞
LI Zhen;YAO Hong-yu(Department of Colorectal and Anal Surgery,the First Afliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《医药论坛杂志》
2021年第5期14-18,共5页
Journal of Medical Forum
基金
2018年度河南省高等学校重点科研项目(18A320058)。
关键词
微卫星不稳定性
错配修复
散发性结直肠癌
病理特点
Microsatellite Instability
Mismatch Repair
Sporadic Colorectal Cancer
Clinicopathological Features
