PAX1甲基化与蛋白检测在宫颈鳞癌及上皮内病变中的应用价值分析

Analysis on the application value of PAX1 methylation and protein detection in cervical squamous cell carcinoma and intraepithelial lesions

目的分析配对盒家族基因1(Paired boxed gene 1,PAX1)的甲基化检测与PAX1蛋白在宫颈鳞癌及上皮内病变中的阳性率及癌组的临床病理学意义。方法选取2014年1月至2018年10月在遵义医科大学附属医院确诊为宫颈病变的320例患者为研究对象,根据病理诊断结果将其分为慢性宫颈炎(组Ⅰ,93例)、低级别鳞状上皮内病变(组Ⅱ,20例),高级别鳞状上皮内病变(组Ⅲ,112例)及宫颈鳞癌(组Ⅳ,95例),并收集各组患者宫颈组织石蜡标本。采用实时荧光定量聚合酶链反应检测PAX1甲基化情况作定性分析,运用免疫组织化学Envision法检测PAX1蛋白表达情况,利用同类PAX蛋白判读标准进行定性分析,分析不同分组的甲基化率、蛋白表达阳性率以及癌组的临床病理参数特征有无差异。结果4组患者PAX1甲基化率分别为1.08%(1/93)、25.00%(5/20)、66.07%(74/112)、89.47%(85/95),4组及组间两两比较均具有显著统计学差异(P<0.05);组Ⅳ,PAX1甲基化率与患者复发相关,差异具有统计学意义(P<0.05),与年龄、病理分型、淋巴结转移的病理特征无明显差异性(P>0.05);PAX1蛋白阳性率分别为1.08%(1/93)、70.00%(14/20)、96.43%(108/112)、89.47%(85/95),4组比较具有统计学差异(P<0.05),两两比较,组Ⅳ、组Ⅲ、组Ⅱ分别与组Ⅰ,组Ⅲ与组Ⅱ差异有统计学意义(P<0.05);组Ⅳ,PAX1蛋白的表达与年龄、病理分型、淋巴结转移、复发病理特征无明显差异性(P>0.05);4组患者总PAX1甲基化率、蛋白阳性率差异具有统计学意义(P<0.05),两种检测方法之间存在关联性,其关联强度?=0.338。结论①PAX1甲基化率随宫颈病变程度加重逐渐增高,且与宫颈鳞癌复发相关,推测PAX1甲基化可能与患者疾病进展相关联,有助于判断病变预后情况;②PAX1蛋白检测有助于宫颈高级别与低级别鳞状上皮内病变的鉴别,且在宫颈鳞癌的表达与临床病理特征无明显关联;③PAX1甲基化与PAX1蛋白表达随宫颈病变程度的加重总体呈上升趋势,PAX1甲基化与其蛋白表达呈正相关。

Objective To analyze the positive rate and clinic pathological significance of paired boxed gene 1(PAX1)methylation detection and PAX1 protein in cervical squamous cell carcinoma and intraepithelial lesions of the cancer group.Methods From January,2014 to October,2018,in the Affiliated Hospital of Zunyi Medical University,a total of 320 patients diagnosed with cervical lesions was selected as research objects.According to the pathological diagnosis,this study was divided into four groups as follows:chronic cervicitis group(named as Group I,including 93 cases),and low grade squamous intraepithelial lesion group(GroupⅡ,20 cases),high-grade squamous intraepithelial lesion group(GroupⅢ,112 cases)and cervical squamous carcinoma group(Group Ⅳ,95 cases).Paraffin specimens of cervical tissue from each group were collected.Quantitative real-time fluorescence polymerase chain reaction(qrt-pcr)was used to detect PAX1 methylation for qualitative analysis.Immunohistochemistry Envision method was applied to detect PAX1 protein expression and the same PAX protein interpretation standard was used for qualitative analysis on the statistical significance between the methylation rates and positive rates of protein expression and on that of their clinical pathological parameters in the carcinoma groups.Results The PAX1 methylation rate in the four groups was 1.08%(1/93),25%(5/20),66.07%(74/112),and 89.47%(85/95),respectively.In Group IV,PAX1 methylation rate was associated with recurrence of patients,but it had no correlation with such clinical pathological features as patients age,pathological type and lymph node metastasis.The positive rate of PAX1 protein in the four groups was 1.08%(1/93),70.00%(14/20),96.43%(108/112),and 89.47%(85/95),respectively.According to pairwise comparison,there was statistically significant difference between GroupⅣ/Ⅲ/Ⅱand Group,and between GroupⅢand GroupⅡ.In Group IV,PAX1 protein expression had no correlation with such pathological features as patients age,pathological type,lymph node metastasis and recurrence.The difference between the total PAX1 methylation rate and the positive protein rate had statistical significance.Correlation existed between the two detection methods and the correlation intensity was 0.338.Conclusion(1)The methylation rate of PAX1 increases gradually with the deepening of cervical lesions,and is related to the recurrence of cervical squamous cell carcinoma.This finding suggested that PAX1 methylation might be associated with disease progression in patients,which may be helpful in predicting the prognosis of lesions;(2)The detection of PAX1 protein is helpful for the identification of high and low cervical squamous intraepithelial lesions,and the expression of PAX1 protein in cervical squamous carcinoma is not significantly correlated with the clinic pathological features;(3)PAX1 methylation and PAX1 protein expression are increased along with the aggravation of cervical lesions,and PAX1 methylation is positively correlated with its protein expression.