目的 探讨Ki-67、微小染色体维持蛋白2（MCM2）、p16在宫颈鳞状上皮内病变中的表达及其临床意义。方法 采用免疫组化检测Ki-67、MCM2、p16在宫颈炎症组14例、低级别鳞状上皮内病变（LSIL）组47例、高级别鳞状上皮内病变（HSIL）组49例中的表达情况,以病理结果作为金标准,对结果进行统计分析。结果 HSIL组中Ki-67、MCM2、p16阳性率均高于炎症组和LSIL组（均P<0.017）。HSIL组中Ki-67、MCM2过表达率均显著高于炎症组和LSIL组（均P<0.017）。随着宫颈病变级别增加,Ki-67及MCM2阳性范围从基底层至表层逐渐扩大。MCM2及Ki-67在LSIL组中表达模式多为基底层的非过表达模式,HSIL组多为中层及以上的过表达模式。Spearman相关性分析显示,MCM2和Ki-67在宫颈鳞状上皮内病变中的表达强度之间呈正相关（r=0.801,P<0.05）;p16与MCM2在宫颈鳞状上皮内病变中的表达呈正相关（r=0.559,P<0.05）;p16与Ki-67在宫颈鳞状上皮内病变中的表达呈正相关（r=0.478,P<0.05）。结论 p16阳性提示宫颈高级别鳞状上皮内病变。MCM2与Ki-67在宫颈鳞状上皮内病变中的表达具有较高一致性,MCM2可作为宫颈鳞状上皮内病变新的增殖标志物。

Objective To investigate the expression and significance of Ki-67,MCM2 and p16 in cervical intraepithelial lesions．Methods The expressions of Ki-67,MCM2 and p16 in cervicitis group（14 cases）,low-grade squamous intraepithelial lesion（LSIL） group（47 cases）and high-grade squamous intraepithelial lesion（HSIL） group（49 cases）were detected by immunohistochemistry．The pathological results were used as the gold standard for statistical analysis．Results The positive rates of Ki-67,MCM2 and p16 in HSIL group were significantly higher than those in cervicitis group and LSIL group（P<0.017）．The over-expression rates of Ki-67,MCM2 in HSIL group were significantly higher than those in cervicitis group and LSIL group（P<0.017）．With the increase of cervical lesion grade,the positive range of Ki-67 and MCM2 gradually expanded from basal layer to surface layer．The expression patterns of MCM2 and Ki-67 in LSIL group were mostly non-overexpressed in the basal layers,while those in HSIL group were mostly overexpressed in the middle layer and above．Spearman correlation analysis showed that the expression intensity of MCM2 and Ki-67 in cervical squamous intraepithelial lesions was positively correlated（r=0.801,P<0.05）．There was a positive correlation between the expression of p16 and MCM2 in cervical squamous intraepithelial lesions（r=0.559,P<0.05）．There was a positive correlation between the expression of p16 and Ki-67 in cervical squamous intraepithelial lesions（r=0.478,P<0.05）．Conclusions Positive p16 indicates high-grade squamous intraepithelial lesion．The expression of MCM2 and Ki-67 in cervical intraepithelial lesions is highly consistent．MCM2 can be used as a new proliferative marker for cervical intraepithelial lesions．

目的 探讨p16免疫细胞化学、人乳头瘤病毒(HPV)DNA基因分型单独和联合检测在宫颈细胞学不能明确诊断意义的非典型鳞状上皮细胞(ASC-US)分流中的价值。方法 收集2017年3月—2022年1月,585例液基薄层细胞学(TCT)诊断为ASC-US患者的宫颈细胞学标本,使用免疫细胞化学法行p16蛋白检测,生物芯片法行HPV DNA基因分型检测,患者于8周内行阴道镜下病理活检术。以组织学诊断结果为金标准,探讨p16免疫细胞化学和HPV DNA基因分型单独和联合检测方法在同一级别宫颈病变中阳性率的差异,对比同一检测方法在不同级别宫颈病变中阳性率的差异,比较p16免疫细胞化学、HPV DNA基因分型单独和联合检测对高级别鳞状上皮内病变(HSIL)以上病变诊断效能的差异,综合评定一种最优的ASC-US分流方法。结果 ①(p16+HPV DNA)联合检测在同一级别宫颈病变中阳性率高于p16免疫细胞化学、HPV DNA基因分型检测。②p16免疫细胞化学、HPV DNA基因分型、(p16+HPV DNA)联合检测的阳性率均随着宫颈病变程度的加重而递增。③(p16+HPV DNA)联合检测的综合诊断效能最佳,其灵敏度、特异度、符合率和约登指数分别为99.07%、62.55%、69.23%、0.62。结论 p16免疫细胞化学检测法与HPV DNA基因分型单独和联合检测均有助于ASC-US分流,但是,(p16+HPV DNA)联合检测具有更优的灵敏度和约登指数,同时保持了较高的特异度和符合率,可有效进行ASC-US分流。

Objective To investigate the value of p16 immunocytochemistry and human papillomavirus (HPV) DNA genotyping alone and combined in atypical squamous cells of undetermined significance (ASC-US) shunt which cervical cytology can not clearly diagnose. Methods From March 2017 to January 2022, cervical cytological specimens of 585 patients with ASC-US diagnosed by liquid based thinprep cytology test (TCT) were collected. p16 protein was detected by immunocytochemistry, HPV DNA genotype was detected by biochip and the patients underwent colposcopy pathological biopsy within 8 weeks. Taking the histological diagnosis results as the gold standard, the differences of the positive rate of p16 immunocytochemistry and HPV DNA genotyping in the same level of cervical lesions, differences of the positive rate of the same detection method in different levels of cervical lesions and differenes of p16 immunocytochemistry HPV DNA genotyping alone and combined detection of the diagnostic efficacy of lesions severer than high-grade squamous intraepithelial lesion (HSIL) were compared to comprehensively evaluate an optimal ASC-US shunt method. Results ①The positive rate of combined detection of (p16+HPV DNA) in the same level of cervical lesions was higher than that of differences of p16 immunocytochemistry and HPV DNA genotyping. ②The positive rate of combined detection of (p16+HPV DNA), p16 immunocytochemistry and HPV DNA genotyping increased with the aggravation of cervical lesions. ③The combined detection of (p16+HPV DNA) had the best comprehensive diagnostic efficiency and its sensitivity, specificity, coincidence rate and Yoden index were 99.07%, 62.55%, 69.23% and 0.62 respectively. Conclusions p16 immunocytochemical assay and HPV DNA genotyping, both alone and in combination, contributed to ASC-US shunt. However, the combined detection of (p16+HPV DNA) had better sensitivity and Yoden index, with high specificity and coincidence rate, which can effectively carry out ASC-US shunt.

目的 探讨p16/Ki-67染色、HPV E6/E7mRNA和联合检测对HSIL+病变诊断的临床价值。方法 募集2017年3月—2020年8月期间,于中山市博爱医院妇产科就诊,组织学证实为宫颈炎患者209例、LSIL患者169例、HSIL患者131例和宫颈癌患者86例作为研究对象,回顾分析研究对象术前细胞学样本p16/Ki-67染色、HPV E6/E7mRNA检测结果,纵向比较p16/Ki-67染色、HPV E6/E7mRNA和联合检测在不同级别宫颈病变的阳性率的差异,横向比较p16/Ki-67染色、HPV E6/E7mRNA和联合检测在相同宫颈病变的阳性率的差异,综合评估p16/Ki-67染色、HPV E6/E7mRNA和联合检测诊断HSIL+病变效能的差异。结果 ①纵向比较:p16/Ki-67染色、HPV E6/E7mRNA和联合检测阳性率随宫颈病变程度的加重呈趋势性升高(p16/Ki-67染色:χ²=374.34,P<0.001;HPV E6/E7mRNA检测:χ²=289.21,P<0.001;联合检测:χ²=343.90,P<0.001)。②横向比较:在宫颈炎、LSIL、宫颈癌组,p16/Ki-67染色、HPV E6/E7mRNA和联合检测阳性率之间差异均不具有统计学意义(均P>0.05)。在HSIL组,p16/Ki-67染色和联合检测之间阳性率差异有统计学意义(χ²=8.09,P=0.004); HPV E6/E7mRNA和联合检测之间阳性率差异有统计学意义(χ²=11.30,P=0.001)。③p16/Ki-67染色、HPV E6/E7mRNA和联合检测诊断HSIL+的灵敏度,总体差异有统计学意义(χ²=7.69,P=0.021)。p16/Ki-67染色与联合检测法之间的灵敏度差异有统计学意义(χ²=7.29,P=0.007);HPV E6/E7mRNA检测与联合检测法之间的灵敏度差异有统计学意义(χ²=4.84,P=0.028)。p16/Ki-67染色、HPV E6/E7mRNA和联合检测诊断HSIL+的特异度及符合率的总体差异不具有统计学意义(χ²₁=5.38,P₁=0.068;χ²₂=0.93,P₂=0.628)。结论 p16/Ki-67染色、HPV E6/E7mRNA 和联合检测均可有效筛出HSIL+病变,但是联合检测能显著提高HSIL+病变诊断的灵敏度,降低漏诊率,同时保持了较好的特异度和符合率,建议将p16/Ki-67染色和HPV E6/E7mRNA联合检测作为早期诊断HSIL+病变的策略。

Objective To investigate the clinical value of p16/Ki-67 staining E6/E7 mRNA and combined detection in the diagnosis of HSIL+ lesions. Methods From March 2017 to August 2020,209 cases of cervicitis,169 cases of LSIL,131 cases of HSIL and 86 cases of cervical cancer confirmed by histology were selected as the research objects. The results of p16/Ki-67 staining and HPV E6/E7 RNA detection of the preoperative cytological samples were retrospectively analyzed and the p16/Ki-67 staining and HPV E6/E7 mRNA detection results were compared longitudinally. The positive rates of E6/E7mRNA and combined detection in different grades of cervical lesions were compared.The positive rates of p16/Ki-67 staining,HPV E6/E7 mRNA and combined detection in the same cervical lesions were compared horizontally.The differences in the diagnostic efficacy of p16/Ki-67 staining,HPV E6/E7 mRNA and combined detection in the diagnosis of HSIL+lesions were comprehensively evaluated. Results ①Longitudinal comparison:the positive rates of p16/ Ki-67 staining, HPV E6/E7mRNA and combined detection increased with the severity of cervical lesions(p16/Ki-67 staining:χ²=374.34,P<0.001;HPV E6/E7 mRNA detection:χ²=289.21,P<0.001;joint detection:χ²=343.90,P<0.001). ②Transverse comparison: in cervicitis, LSIL and cervical cancer groups,there were no significant differences in the positive rates of p16/Ki-67 staining, HPV E6/E7 mRNA and combined detection (all P>0.05). In the HSIL group,there was significant difference in the positive rate between p16/Ki-67 staining and combined detection (χ²=8.09,P=0.004)and the difference between HPV E6/ E7 mRNA and combined detection was statistically significant(χ²=11.30,P=0.001). ③The sensitivity of p16/Ki-67staining,HPV E6/E7 mRNA and combined detection in the diagnosis of HSIL+ was statistically significant(χ²=7.69,P=0.021). The sensitivity difference between p16/Ki-67 staining and combined detection was statistically significant(χ²=7.29,P=0.007);the sensitivity difference between HPV E6/E7 mRNA detection and combined detection method was statistically significant (χ²=4.84,P=0.028). There was no significant difference in the specificity and coincidence rate of p16/Ki-67 staining,HPV E6/E7 mRNA and combined detection in the diagnosis of HSIL+(χ²₁=5.38,P₁=0.068;χ²₂=0.93,P₂=0.628). Conclusion P16/Ki-67 staining,HPV E6/E7 mRNA and combined detection may effectively screen out HSIL+ lesions,reduce the missed diagnosis rate, but the combined detection may significantly improve the sensitivity of diagnosis of HSIL+ lesions, while maintaining good specificity and coincidence rate.It is suggested that p16/Ki-67 staining and HPV E6/E7 mRNA detection should be used as a strategy for early diagnosis of HSIL+ lesions.

目的 研究细胞周期调控因子E2F-1和p16蛋白与宫颈上皮内瘤变及宫颈鳞癌的表达及其临床意义。方法 通过免疫组化SP法检测宫颈上皮内瘤变Ⅰ-Ⅱ级(CINⅠ-Ⅱ级)、宫颈上皮内瘤变Ⅲ级(CINⅢ级)、宫颈鳞癌各40例中E2F-1和p16蛋白的表达情况,并用20例慢性宫颈炎组织作对照。结果 E2F-1蛋白在CINⅠ-Ⅱ级、CINⅢ级和宫颈鳞癌组中阳性率分别为17.5%、67.5%和80.0%, 3组间有差异(χ²=37.278,P<0.001)。p16蛋白表达阳性率分别为27.5%、70.0%和82.5%,3组间差异有统计学意义(χ²=27.708, P<0.001)。CINⅠ-Ⅱ级与CINⅢ级组及宫颈鳞癌组比较均有差异,但CINⅢ级与宫颈鳞癌组间无差异。p16蛋白的表达与E2F-1蛋白表达有正相关关系;在鳞癌组中E2F-1与p16的异常表达与组织学分级、临床分期有关(P<0.05)。结论 子宫颈鳞癌的形成与E2F-1 、p16蛋白过表达是呈正相关关系,E2F-1 、p16 蛋白可能作为子宫颈鳞癌及CIN的标志物,对子宫颈癌筛查和预防有重要意义。

Objective To explore the clinicopathologic significance of the protein expression of cell cycle regulators E2F-1 and P16 in cervical intraepithelial neoplasia(CIN) and cervical squamous cell carcinoma(SCC). Methods Immunohistochemical technique S-P was used to determinate the expression of E2F-1 and P16 protein in 40 cases with CINⅠ-Ⅱ, 40 cases with CINⅢ and 40 cases with cervical squamous cell carcinoma. Results The positive rate of E2F-1 protein expression in CINⅠ-Ⅱ, CINⅢ and cervical squamous cell carcinoma was 17.5%,67.5% and 80.0%, respectively. There were significant differences among the three groups(χ²=37.278,P<0.001). The positive rate of P16 protein expression in CINⅠ-Ⅱ, CINⅢ and cervical squamous cell carcinoma was 27.5%,70.0% and 82.5%, respectively. There were significant differences among the three groups (χ²=27.708, P<0.001). The group with CINⅢ and cervical squamous cell carcinom compared with the group withCINⅠ-Ⅱ did have differences. But there were no significant differences in the group with CINⅢ and the group with cervical squamous cell carcinoma. The relative expressions between P16 and E2F-1 showed positive correlation. In the cervical squamous cell carcinoma group, the abnormal expression of E2F-1 and p16 were correlated with the histological grade and clinical stage. Conclusion It showed positive correlation between very high expression of p16 protein and E2F-1 infection in CIN and SCC. p16 protein and E2F-1 can be an indicator in SCC and CIN,which has very significance in preventing and screening of cervical cancer.

       目的   探讨Ki-67、微小染色体维持蛋白2（MCM2）、p16在宫颈鳞状上皮内病变中的表达及其临床意义。方法   采用免疫组化检测Ki-67、MCM2、p16在宫颈炎症组14例、低级别鳞状上皮内病变（LSIL）组47例、高级别鳞状上皮内病变（HSIL）组49例中的表达情况，以病理结果作为金标准，对结果进行统计分析。结果  HSIL组中Ki-67、MCM2、p16阳性率均高于炎症组和LSIL组（均P＜0.017）。HSIL组中Ki-67、MCM2过表达率均显著高于炎症组和LSIL组（均P＜0.017）。随着宫颈病变级别增加，Ki-67及MCM2阳性范围从基底层至表层逐渐扩大。MCM2及Ki-67在LSIL组中表达模式多为基底层的非过表达模式，HSIL组多为中层及以上的过表达模式。Spearman相关性分析显示，MCM2和Ki-67在宫颈鳞状上皮内病变中的表达强度之间呈正相关（r=0.801，P＜0.05）；p16与MCM2在宫颈鳞状上皮内病变中的表达呈正相关（r=0.559，P＜0.05）；p16与Ki-67在宫颈鳞状上皮内病变中的表达呈正相关（r=0.478，P＜0.05）。结论  p16阳性提示宫颈高级别鳞状上皮内病变。MCM2与Ki-67在宫颈鳞状上皮内病变中的表达具有较高一致性，MCM2可作为宫颈鳞状上皮内病变新的增殖标志物。

       Objective  To investigate the expression and significance of Ki-67，MCM2 and p16 in cervical intraepithelial lesions．Methods  The expressions of Ki-67，MCM2 and p16 in cervicitis group（14 cases），low-grade squamous intraepithelial lesion（LSIL） group（47 cases）and high-grade squamous intraepithelial lesion（HSIL） group（49 cases）were detected by immunohistochemistry．The pathological results were used as the gold standard for statistical analysis．Results  The positive  rates of Ki-67，MCM2 and p16 in HSIL group were significantly higher than those in cervicitis group and LSIL group（P＜0.017）．The over-expression rates of Ki-67，MCM2 in HSIL group were significantly higher than those in cervicitis group and LSIL grou（P＜0.017）．With the increase of cervical lesion grade，the positive range of Ki-67 and MCM2 gradually expanded from basallayer to surface layer．The expression patterns of MCM2 and Ki-67 in LSIL group were mostly non-overexpressed in the basal layers，while those in HSIL group were mostly overexpressed in the middle layer and above．Spearman correlation analysis showed that the expression intensity of MCM2 and Ki-67 in cervical squamous intraepithelial lesions was positively correlated（r=0.801，P＜0.05）．There was a positive correlation between the expression of p16 and MCM2 in cervical squamous intraepithelial lesions（r=0.559，P＜0.05）．There was a positive correlation between the expression of p16 and Ki-67 in cervical squamous intraepithelial lesions（r=0.478，P＜0.05）．Conclusions  Positive p16 indicates high-grade squamous intraepithelial lesion．The expression of MCM2 and Ki-67 in cervical intraepithelial lesions is highly consistent．MCM2 can be used as a new proliferative marker for cervical intraepithelial lesions．