一种宫颈鳞癌瘤芽评估的新方法

A new method for evaluation of cervical squamous cell carcinoma tumor budding

目的 探索一种一致性高的宫颈鳞癌瘤芽评估方法。方法 比较两名病理医师筛选肿瘤浸润最深切片及瘤芽密度最大切片的一致性,并筛选出一致性高的瘤芽评估对象;选择一致性高的瘤芽评估对象,由两名病理医师分别计数HE染色切片单视野最高瘤芽数及10个视野平均瘤芽数,采用组内相关系数(ICC)评估计数结果的一致性;选择一致性高的瘤芽评估对象与计数方法对95例份宫颈鳞癌组织HE染色切片重新进行瘤芽评估,绘制受试者工作特征(ROC)曲线,获取最佳截断值,并依此分为高瘤芽组与低瘤芽组,分析瘤芽数对宫颈鳞癌脉管侵犯的预测效能以及瘤芽数与脉管侵犯的关系。取宫颈鳞癌组织蜡块,通过免疫组化染色观察CK、p63、p40染色情况。结果 筛选肿瘤浸润最深切片的一致率高于瘤芽密度最大切片,但二者比较差异无统计学意义(P>0.05);两名病理医师计数瘤芽密度最大切片单视野最高瘤芽数的ICC为0.769,计数10个视野平均瘤芽数的ICC为0.346。瘤芽密度最大切片单视野最高瘤芽数预测宫颈鳞癌脉管侵犯的ROC曲线下面积为0.736(95%CI:0.636~0.836),其最佳截断值为2.5个,此时其预测宫颈鳞癌脉管侵犯的灵敏度为62.8%、特异度为73.1%。根据瘤芽密度最大切片单视野最高瘤芽数将95例宫颈鳞癌患者分为高瘤芽组44例、低瘤芽组51例。高瘤芽组脉管侵犯的发生率显著高于低瘤芽组(P<0.05)。Spearman相关分析显示,宫颈鳞癌患者脉管侵犯与瘤芽数呈正相关关系(r=0.466,P<0.01)。瘤芽与无核细胞微滴CK染色均呈阳性,而部分肿瘤细胞p63、p40表达缺失。结论 通过HE染色评估瘤芽密度最大切片单视野最高瘤芽数是简单易行、重复性较高的宫颈鳞癌瘤芽评估方法。

Objective To explore a highly consistent method for evaluating the tumor budding of cervical squamous cell carcinoma.Methods The consistency of the sections with the deepest tumor invasion and those with the highest tumor budding density screened by two pathologists was compared,and the tumor budding evaluation objective with high consistency was selected.The highest number of tumor buddings in single visual field and the average number of tumor buddings in 10 visual fields in HE staining sections were counted by two pathologists using the evaluation objective with high consistency,respectively.The consistency of the counting results was evaluated using intra-class correlation coefficient(ICC).Re-budding evaluation was carried out in 95 HE staining sections of cervical squamous cell carcinoma tissues using the evaluation objective and the counting method with high consistency.The receiver operating characteristic(ROC) curve was drawn and the optimal cut-off was obtained.Based on this,the patients were divided into the high-grade tumor budding group and low-grade tumor budding group.The predictive effect of tumor budding count on vascular invasion in cervical squamous cell carcinoma as well as the correlation between tumor budding count and vascular invasion were analyzed.The paraffin blocks of cervical squamous cell carcinoma tissues were taken,and CK,p63 and P40 were observed by immunohistochemical staining.Results The consistency rate of the screened sections with the deepest tumor invasion was higher than that of the sections with the highest tumor budding density,without statistically significant difference(P>0.05).The ICC of the two pathologists for counting the largest number of tumor buddings in single visual field in the sections with the highest tumor budding density was 0.769,and that for counting the average number of tumor buddings in 10 visual fields was 0.346.The area under the ROC curve(AUC) of the largest number of tumor buddings in the single visual fields in the sections with the highest tumor budding density in predicting vascular invasion of cervical squamous cell carcinoma was 0.736(95% CI:0.636-0.836),and the optimal cut-off was 2.5;the sensitivity and specificity were 62.8%and 73.1%,respectively.Based on the largest number of tumor buddings in single visual fields in the sections with the highest tumor budding density,95 patients with cervical squamous cell carcinoma were divided into the high-grade tumor budding group(n=44) and low-grade tumor budding group(n=51).The incidence of vascular invasion in the highgrade tumor budding group was significantly higher than that in the low-grade tumor budding group(P<0.05).Spearman correlation analysis showed a positive correlation between vascular invasion and tumor bud count in patients with cervical squamous cell carcinoma(r=0.466,P<0.01).Both tumor buddings and anucleate cells presented CK-positive in droplet staining,while some tumor cells showed loss of p63 and p40 expression.Conclusion Evaluating the largest number of tumor buddings in single visual fields in the sections with the highest tumor budding density through HE staining is a simple and highly reproducible method to evaluate the tumor budding of cervical squamous cell carcinoma.