The Association of Molecular Biomarkers in the Diagnosis of Cervical Pre-Cancer and Cancer and Risk Factors in Senegalese

Background: Cervical intraepithelial neoplasia (CIN) grading is subjective and affected by substantial rates of discordance among pathologists. Although recent studies have suggested that p16INK4a may be a useful surrogate biomarker of cervical neoplasia, Ki-67 and human papillomavirus testing have also been shown to be useful in detecting neoplasia. The purpose of this study was to determine the expression of p16INK4a and Ki-67 in cervical neoplasia and its correlations with cofactors. Methods: The study involved 69 patients with and without cervical neoplasia who underwent colposcopic directed biopsy. On each patient, two samples were taken; the first was used for immunohistochemistry and the second for molecular testing, using HPV16and18 genotyping Real-Time PCR Kit. Results: The study revealed the expression level of p16INK4a and Ki-67 in a descending order, from invasive squamous cell carcinoma (SCC), CIN2/3, CIN1 and non-dysplastic lesions. Correlations showed an association between the staining of p16NK4a and Ki-67 with the increase of age (OR: 1.79 (95%IC: 0.49 – 6.55), p = 0.037) and marital status (OR: 0.17 (95%IC: 0.04 – 0.68), p = 0.003). We found that the expressions of p16INK4a and Ki-67 were significantly different between invasive SCC vs non-dysplasia (OR: 44.57 (95%IC: 4.91 – 403.91), p <0.0001). The study showed significant correlation between HPV 16and18 infection with p16 INK4a and Ki-67 expression (OR: 0.13 (95%IC: 0.03 – 0.52), p <0.0001). Strong expression of p16INK4a and Ki-67 were observed in invasive squamous cell carcinoma, moderate staining was found in CIN2/3, weak staining in CIN1 and normal histology. Conclusion: Our findings indicate that p16INK4a and Ki-67 expressions associated strongly with cervical pathology. Therefore, p16/Ki-67 could be considered as a suitable biomarker for cervical cancer screening, particularly in HPV-based screening programs.     

Predictive Value of Marker of Proliferation Ki-67 and Cell Cycle Dependent Protein kinase Inhibitor P16INK4a in Cervical Biopsy to Determine Its Biological Behaviour

Objective: The aim of the study is to analyse the Immuno-histochemical expression of   Ki-67  and P16INK4a in CIN and cervical cancer cases  and  their utility to determine the accuracy of histological diagnosis and prediction of biological behavior of cervical lesion. Methodology: A retrospective cross-sectional study was carried in 110 numbers of cervical biopsy that included 25 CIN1, 21 CIN2, 12 CIN3, 26 SCC and 01 adenocarcinoma and 25 non neoplastic lesion. The tissue sections were stained with Ki-67 and P16INK4a. Results: Ki-67 expression was seen in 55.5% (61/110) cases of cervical lesion., out of which 3.6% (4/110; cervicitis -2/110 and metaplasia-2/110) cases were non dysplaia, 51.8% (57/110) cases were dysplasia /CIN of varying grade including invasive cancer. P16INK4a expression was noted 51.8% (57/110). There was an increasing trend of the intensity of Ki-67 and P16INK4a from focal positivity in low grade lesion to diffuse intensity in higher grade lesion and is statistically significant. There was strong association between the two variables Ki-67 and P16INK4a positive cases with their histologic grade. Conclusion: Though histopathology remains the ‘‘gold standard’’ for the diagnosis of CIN, both low and high-grade, biomarkers like Ki-67 and P16INK4a have emerged as helpful adjuncts. Their combined use may assist in the histopathologic classification of preinvasive lesions and facilitate the distinction from nondysplasia.     

P16INK4a和Ki-67在宫颈癌前病变诊断中的意义

目的研究P16INK4a、Ki-67在宫颈癌前病变中的表达及早期诊断价值。方法宫颈活检标本92例,采用免疫组织化学PV-9000二步法检测宫颈鳞癌28例,CIN 45例（CINⅢ级17例、CINⅡ级7例、CINⅠ级21例）,炎性病变19例。结果在宫颈癌、CIN病变、宫颈炎P16INK4a的阳性表达率分别为100%、44.4%和10.5%,Ki-67的阳性表达率分别为100%、51.11%、21.05%,三组不同病变比较均差异有显著性（P〈0.001）。P16INK4a与Ki-67表达存在正相关（r2=0.893,P〈0.001）。结论 P16INk4a、Ki-67联合检测可以提高宫颈癌的早期诊断率。     

P16INK4a及Ki-67免疫细胞化学检测对ASC-US中高级别病变检出的意义

目的：探讨液基细胞学剩余细胞标本P16INK4a、Ki-67免疫细胞化学染色检测宫颈癌前病变的价值。方法：选取2010年1月至2012年6月因宫颈疾病于北京大学第三医院妇产科就诊患者液基细胞学剩余标本50例。所有患者行高危型HPVDNA杂交捕获II代（HCII）检测,P16INK4a、Ki-67免疫细胞化学检测,同时行阴道镜检查及组织病理学活检。结果：以病理诊断将患者分为CIN2以下组和CIN2及以上组。CIN2及以上组P16INK4a及Ki-67表达量高于CIN2以下组,差异均具有统计学意义（P〈0．05）;P16INK4a或Ki-67检测对高级别病变预测的准确性在ASC—US组中优于异常细胞学组;在ASC—US中,P16INK4a或Ki-67检测对高级别病变预测的准确性优于高危型HPV检测。结论：宫颈脱落细胞中P16INK4a或Ki-67免疫细胞化学检测相比于高危型HPV检测可以提高对ASC—US中高级别病变的检出作用。     

p16和Ki-67免疫组化检测与宫颈上皮内瘤变(CIN)程度的相关性分析

目的:分析免疫组化检测p16和Ki-67与宫颈上皮内瘤变（CIN）严重程度的关系,以及 p16联合Ki-67在高级别CIN鉴别中的效能。方法:采用回顾性研究,纳入2016年1月至2018年3月期间粤北人民医院经HE染色病理确诊的宫颈病变患者160例,其中慢性宫颈炎者共38例归为对照组,CINⅠ级、Ⅱ级、Ⅲ级分别为41例、46例、35例;再将CIN患者分为低级别鳞状上皮内病变（LSIL）和高级别鳞状上皮内病变（HSIL）组。对比对照组与CINⅠ级、Ⅱ级、Ⅲ级组、ISIL组、HSIL组p16及Ki-67阳性表达情况;采用Spearman相关性分析观察p16阳性表达、Ki-67阳性表达与CIN严重程度的关系。联合使用p16、Ki-67半定量评分,评估联合p16、Ki-67半定量评分诊断结果与病理结果一致性,评估效能。结果:CINⅠ-Ⅲ级组p16阳性率较对照组高,CINⅡ、Ⅲ级p16阳性率较CINⅠ级高;HSIL组阳性率较LSIL组高,差异均有统计学意义,P＜0.05。Ki-67在对照组中阳性率为0.00%,在LSIL组、HSIL组中分别为68.29%、100.00%。p16与宫颈病变程度呈正相关,r=0.692,P=0.021 0;Ki-67与宫颈病变程度呈正相关,r=0.612,P=0.001 8。p16联合Ki-67诊断炎症结果与病理完全一致,CINⅠ级共36例（87.80%）符合,CINⅡ级共43例（93.48%）符合,CINⅢ级共34例（97.14%）符合;LSIL符合率为87.80%（36/41）,HSIL符合率为95.06%（77/81）。p16联合Ki-67诊断LSIL敏感性、特异性为94.74%和97.58%,诊断HSIL敏感性、特异性分别为96.25%、98.81%,诊断CIN敏感性、特异性分别为92.62%、96.48%,其中LSIL敏感性高于单用,CIN诊断特异性较单用p16高,P＜0.05。结论:p16和Ki-67表达与宫颈上皮内瘤样病变严重程度密切相关,可以通过观察二者表达情况辅助CIN病情严重程度判断,联合p16和Ki-67免疫组化检测能提高诊断高级别鳞状上皮内病变诊断特异性、敏感性。     

ｐ１６和Ｋｉ-６７蛋白在宫颈上皮内瘤变中的临床意义

目的　研究ｐ１６和Ｋｉ-６７蛋白在宫颈组织中的表达,探讨它们在宫颈病变中的诊断价值。方法　应用免疫组化技术ＳＰ法检测正常宫颈或炎性病变组织２０例、ＣＩＮⅠ组织２２例、ＣＩＮⅡ组织２０例、ＣＩＮⅢ组织２０例、宫颈腺体受累及组织２０例（未见原发病灶）、宫颈鳞状细胞癌组织２０例中ｐ１６和Ｋｉ-６７蛋白的表达情况。结果　ｐ１６和Ｋｉ-６７蛋白在正常宫颈组织中的阳性表达率明显低于宫颈癌前病变及宫颈癌组织,差异有显著意义（Ｐ＜０.０１）。Ｋｉ-６７蛋白阳性表达在良性宫颈组织与ＣＩＮⅠ组织相比较,无显著差异（Ｐ＞０.０５）;ｐ１６阳性表达在宫颈基底细胞增生组织与ＣＩＮⅠ组织相比较,有显著差异（Ｐ＜０.０５）,而Ｋｉ-６７蛋白无显著差异（Ｐ＞０.０５）;ｐ１６着色强阳性在ＣＩＮⅠ与ＣＩＮⅡ、ＣＩＮⅢ、宫颈癌比较,有显著差异（Ｐ＜０.０１）。结论　ｐ１６蛋白能很好地鉴别宫颈良性病变、宫颈上皮内瘤变以及宫颈癌;在鉴别宫颈基底细胞增生与ＣＩＮⅠ时,ｐ１６能起辅助诊断作用;应用ｐ１６的着色强弱能够很好地区分ＣＩＮⅠ与ＣＩＮⅡ、ＣＩＮⅢ,有助于ＣＩＮ的判断与分级。Ｋｉ-６７能很好地区分宫颈高度病变与宫颈癌,但不能区分宫颈良性病变与宫颈低度病变,需结合ｐ１６蛋白联合判断。     

Application of biomarkers in the diagnosis of cervical precancerous lesions

Several molecular biomarkers (p16INK4a, Ki-67, and so on) are abnormally expressed in cervical precancerous lesions, many of which are involved in human papilloma virus (HPV)-induced molecular alterations. Cervical intraepithelial neoplasia (CIN) can not accurately predict the possibility of progress in precancerous lesions. Atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesions (LSIL) are often over-treated clinically, while it lacks accurate screening methods to identify the precancerous lesions with high possibility of progression. Combining cytology diagnosis with biomarkers can help predict the progress of precancerous lesions and distinguish benign lesions which are difficult to identify morphologically. It appears promising to consider biomarkers as potential cervical precancerous lesions screening method.     

宫颈上皮内瘤样病变发生发展与p16/Ki-67共表达的相关性研究进展

宫颈上皮内瘤样病变（cervical intraepithelial neoplasia,CIN）属于宫颈癌前病变,虽然CIN进展成宫颈癌时间一般较长,但其发病率增多及发病年龄变小,对女性身心健康造成严重影响。随着宫颈癌筛查技术的进步和分子生物学技术的发展,高级别的宫颈癌上皮内瘤样变的早期检查确认并进行阻断已成为筛查的目标。p16被认为是发现最早的抑癌基因,在鉴别低级别与高级别宫颈鳞状上皮内病变中有重要作用。Ki-67为常见肿瘤标志物,在多种肿瘤中表达。p16/Ki-67双染为新型宫颈病变筛查或诊断方法,即同一细胞内检测到p16、Ki-67共表达,则提示细胞周期失调,考虑为宫颈上皮内高级别瘤样病变。国内近年有研究报道p16和Ki-67在CIN中的作用,未见到相关的综述报道。本文对宫颈上皮内瘤样病变与p16/Ki-67共表达的相关性作一综述,为临床宫颈病变鉴别、诊断提供参考。     

The utility of p16INK4a and Ki-67 staining on cell blocks prepared from residual thin-layer cervicovaginal material

BACKGROUND: Cell blocks can be prepared from residual thin-layer cervicovaginal (ThinPrep) material and can be used in immunohistochemical staining assays for p16INK4a and Ki-67, which are surrogate markers related to human papillomavirus infection and cell proliferation, respectively. The objectives of the current study were 1) to investigate the feasibility and the role of cell block preparations in identifying significant neoplastic and preneoplastic lesions of the uterine cervix and 2) to assess the feasibility of using p16INK4a and Ki-67 immunohistochemical staining patterns on cell blocks to identify significant preneoplastic cervical lesions. METHODS: Cervicovaginal cytology specimens from 85 patients were analyzed. Cytologic diagnoses based on ThinPrep Papanicolaou test results were as follows: squamous cell carcinoma was diagnosed in 3 specimens, high-grade squamous intraepithelial lesions (HSIL) were diagnosed in 27 specimens, low-grade squamous intraepithelial lesions (LSIL) were diagnosed in 20 specimens, and atypical squamous cells of uncertain significance (ASCUS) were diagnosed in 11 specimens. Diagnoses of negativity for intraepithelial lesions or malignancy (NILM) were made in 24 specimens. Cell block sections were stained with hematoxylin and eosin and were immunostained with antibodies against p16INK4a protein and Ki-67 antigen. RESULTS: The cytomorphologic diagnoses made using cell block preparations were as follows: SCC in 2 specimens, HSIL in 20 specimens, LSIL in 30 specimens, NILM in 32 specimens, and no diagnosis in 1 specimen. In 62 cases (73%), the diagnoses made using cell block preparations were in agreement with the ThinPrep diagnoses. Immunostaining of cell blocks for p16INK4a and Ki-67 exhibited a statistically significant association (P < 0.05) with the presence of significant lesions on either cell block or ThinPrep analysis. CONCLUSIONS: To the authors' knowledge, p16INK4a has not been analyzed previously in ThinPrep cell blocks, and the correlation between Ki-67 expression and cell block diagnoses also has not been reported previously. The current results indicate that cell blocks prepared from residual ThinPrep material represent an additional reliable diagnostic tool in the evaluation of cervical samples. Furthermore, immunohistochemical studies may be helpful in differentiating significant preneoplastic changes from other cervical lesions, such as atrophy.     

宫颈上皮内瘤变p16~(INK4a)和p53及Ki-67表达及其诊断价值探讨

目的:探讨用p16和p53及Ki-67等免疫组化标记作为宫颈上皮内瘤变(CIN)诊断的辅助指标。方法:选取我院档案CIN患者133例,年龄20～86岁,中位年龄46岁。经3位高年资病理医师复查确认,其中CIN 1级31例、CIN 2级37例和CIN 3级65例。另选同期我院宫颈慢性炎患者19例,浸润性鳞状细胞癌患者20例作为对照。采用SP法进行p16、p53和Ki-67免疫组化染色,结果独立评分。各级别CIN与免疫组化表达的关系采用单变量χ2检验和Spearman等级相关分析。结果:p16阳性表达位于细胞核或胞核伴胞质,p53和Ki-67定位于细胞核。p16、p53和Ki-67表达阳性率随着CIN的加重而升高,均与CIN级别呈正相关,r值分别为0.789、0.554和0.749,P<0.001。153例CIN 1+标本中p16、p53和Ki-67与组织学诊断符合率分别为96.7%(148/153)、71.9%(110/153)和88.2%(135/153)。在102例CIN 2+标本中,p16、p53和Ki-67免疫组化染色的敏感性分别为97.1%(99/102)、74.5%(76/102)和97.1%(99...     

Expression of the p16INK4a and Ki-67 in relation to the grade of cervical intraepithelial neoplasia and high-risk human papillomavirus infection

Objective

The purposes of this study were to evaluate the expression of p16^INK4a (referred as to p16) and Ki-67 in cervical intraepithelial neoplasia (CIN), and the correlation between high-risk human papillomavirus (HPV) infection and the above biomarkers.

Methods

We analyzed 31 patients who were diagnosed with CIN at Kwandong University Myongji Hospital from October 2006 to September 2007. CIN specimens (CIN1, 12; CIN2, 6; CIN3, 13) were obtained by colposcopy-directed biopsy (CDB) or loop electrical excision procedure (LEEP). The expressions of p16 and Ki-67 were evaluated by immunohistochemical methods with antibodies to p16 and Ki67. The immunohistochemical staining results were classified into four grades: 0, 1, 2 and 3. HPV genotyping or Hybrid Capture-II test was used to detect high-risk HPV.

Results

The expression of p16 (p<0.001) and Ki-67 (p=0.003) were positively associated with CIN grade. p16 expressions increased significantly with high-risk HPV infection (p=0.014), especially HPV type 16 and 58. Ki-67 expression was not related with high-risk HPV. There was positive correlation between the expression of the p16 and Ki-67 (p=0.007).

Conclusion

CIN grade were positively related to the expression of p16 and Ki-67. p16 expressions of high-risk HPV specimens significantly increased more than Ki-67. Therefore, in the diagnosis of CIN and high-risk HPV infection, p16 can be a useful biomarker.     

The role of cyclins and cyclins inhibitors in the multistep process of HPV-associated cervical carcinoma

BACKGROUND: Human papillomavirus (HPV) types 16 and 18 are associated with cervical carcinogenesis. This is possibly achieved through an interaction between HPV oncogenic proteins and some cell cycle regulatory genes. However, the exact pathogenetic mechanisms are not well defined yet. METHODS: We investigated 110 subjects (43 invasive squamous cell carcinoma (ISCC), 38 CIN III, 11 CIN II, 18 CIN I) confirmed to be positive for HPV16 and/or 18 as well as 20 normal cervical tissue (NCT) samples for abnormal expression of cyclin D1, cyclin E, CDK4, cyclin inhibitors (p21 (waf), p27, p16 (INK4A)) and Ki-67 using immunohistochemistry and differential PCR techniques. RESULTS: There was a significant increase in the expression of Ki-67, cyclin E, CDK4, p16 (INK4A) (p=0.003, 0.001, 0.001) and a significant decrease in p27 (Kip1) from NCT to ISCC (p=0.003). There was a significant correlation between altered expression of p27 (KIP1) and p16(INK4A) (p<0.001), cyclin D1 and CDK4 (p=0.001), cyclin E and p27 (Kip1) (p=0.011) in all studied groups. In ISCC, there was significant relationship between standard clinicopathological prognostic factors and high Ki-67 index , increased cyclin D1 and cyclin E, reduced p27 (Kip1) and p21 (waf). CONCLUSION: 1) Aberrations involving p27 (KIP1), cyclin E, CDK4 and p16 (INK4A) are considered early events in HPV 16 and 18-associated cervical carcinogenesis (CINI & II), whereas cyclin D1 aberrations are late events (CINIII & ISCC) 2) Immunohistochemical tests for p16 (INK4A) and cyclin E could help in early diagnosis of cervical carcinoma 3) Only FIGO stage, cyclin D1, p27 (Kip1) and Ki-67 are independent prognostic factors that might help in predicting outcome of cervical cancer patients.     

宫颈上皮内瘤变p16（INK4a）和p53及Ki-67表达及其诊断价值探讨

目的：探讨用p16和p53及Ki-67等免疫组化标记作为宫颈上皮内瘤变（CIN）诊断的辅助指标。方法：选取我院档案CIN患者133例,年龄20～86岁,中位年龄46岁。经3位高年资病理医师复查确认,其中CIN 1级31例、CIN 2级37例和CIN 3级65例。另选同期我院宫颈慢性炎患者19例,浸润性鳞状细胞癌患者20例作为对照。采用SP法进行p16、p53和Ki-67免疫组化染色,结果独立评分。各级别CIN与免疫组化表达的关系采用单变量χ2检验和Spearman等级相关分析。结果：p16阳性表达位于细胞核或胞核伴胞质,p53和Ki-67定位于细胞核。p16、p53和Ki-67表达阳性率随着CIN的加重而升高,均与CIN级别呈正相关,r值分别为0.789、0.554和0.749,P〈0.001。153例CIN 1＋标本中p16、p53和Ki-67与组织学诊断符合率分别为96.7%（148/153）、71.9%（110/153）和88.2%（135/153）。在102例CIN 2＋标本中,p16、p53和Ki-67免疫组化染色的敏感性分别为97.1%（99/102）、74.5%（76/102）和97.1%（99/102）;特异性分别为94.7%（18/19）、84.2%（16/19）和73.7%（14/19）。p16、p53和Ki-67 3者联合在CIN 2＋中的敏感性和特异性分别为98.0%（100/102）、68.4%（13/19）。结论：在宫颈活检的组织学评价时,结合p16、p53和Ki-67免疫组化染色,对提高高级别CIN的检出率、减少漏诊有很大作用。     

Clinical significance of immunocytochemistry for PIK3CA as a carcinogenesis-related marker on liquid-based cytology in cervical intraepithelial neoplasia

The catalytic subunit alpha of phosphatidylinositol 3-kinase (PIK3CA) has been expected to play a role as an important oncogene in uterine cervical carcinoma, whose expression in non-invasive lesions has received considerable attention. We investigated the potential of PIK3CA as a carcinogenesis-related marker for early intraepithelial lesion of the uterine cervix in cytology samples. Seventy-four cases with abnormal cytology suggesting the existence of cervical intraepithelial neoplasia (CIN) lesions, whose findings were histologically confirmed, were selected; they consisted of 20 CIN1, 21 CIN2, and 33 CIN3, respectively. In addition, 17 normal cases, whose cervical cytology revealed no abnormality over three occasions, were selected. Liquid-based cytology specimens were applied for human papillomavirus (HPV) DNA typing and immunocytochemistry using three different antibodies for p16(INK4a), Ki-67 and PIK3CA, respectively. The fraction of immunopositive cells on the slides was calculated and expressed as mean numbers. Receiver operating characteristic (ROC) plots were generated to determine the diagnostic accuracy of each immunocytochemistry test. The mean number of immunopositive cells in the CIN3 group was significantly higher than other groups for all three antibodies. Among all groups, PIK3CA showed a superior specificity to distinguish CIN3 from other groups. Comparison of three markers by ROC curves also revealed that PIK3CA provided the best method for distinguishing CIN3. Thus, expression of PIK3CA was observed in liquid-based cytology in CIN lesions, which suggested its diagnostic significance in addition to the use of routine cervical cancer smear and the HPV screening program.     

p16INK4a Immunoprofiles of Squamous Lesions of the Uterine Cervix–Implications for the Reclassification of Atypical Immature Squamous Metaplasia

p16^INK4a immunoprofiles of non-precancerous and dysplastic squamous cervical lesions were defined and applied to the reclassification of atypical immature squamous metaplasia (AIM). The immunoexpression of cytokeratin 17 (CK 17) in AIM was also evaluated. Totally, 295 cervical cone biopsies representing squamous metaplasia, reactive changes, koilocytosis, flat condyloma, CIN I, CIN II, CIN III and AIM were subjected to p16^INK4a immunohistochemistry. AIM cases were analyzed using CK 17 antibody. Typical p16^INK4a immunoprofiles for the metaplastic, LSIL/HPV and HSIL phenotypes were recorded and used for the categorization of AIM into particular phenotype groups. Results were correlated with CK 17 immunoexpression. All CIN II and CIN III lesions, all but one case of CIN I and all flat condylomas overexpressed p16^INK4a. Other non-precancerous lesions, including koilocytosis, were predominantly negative. Contrary to the sporadic and focal immunostaining, diffuse positivity was associated with the dysplastic features of the lesion. CIN II and CIN III were characterized by a diffuse, strong/weak, full-thickness staining, whereas CIN I showed a heterogeneous diffuse/focal, weak/strong, lower half positivity. One third of AIM lesions may be reclassified as HSIL, one third as LSIL/HPV and one third shows metaplastic phenotype. All AIM cases with metaplastic and LSIL/HPV phenotypes expressed CK 17 diffusely, whereas focal positivity slightly prevailed in AIM with HSIL phenotype. We conclude that p16^INK4a immunohistochemistry is a supporting method for the differential diagnosis of cervical lesions, which may be especially useful for the reclassification of AIM. The efficacy of CK 17 immunohistochemistry seems to be controversial for these purposes.     

Minichromosome maintenance 7 protein is a reliable biological marker for human cervical progressive disease

Objective

This study focused on comparing the expression levels of p16, Ki-67, and minichromosome maintenance 7 (MCM7) protein in normal and affected cervical epithelium to ascertain the biological significance of these markers in detecting progressive cervical disease.

Methods

A quantitative and based on-scanning-microscopy analysis of the three markers expression was performed in normal and cervical intraepithelial neoplasia (CIN) I, II, and III tissues. p16 area as well as p16, Ki-67, and MCM7 positive cells or nuclei were evaluated according to their distribution and extent through the cervical epithelium.

Results

A clear p16 over-expression was observed in all the dysplastic epithelium tissue samples. The quantitative analysis of p16 area as well as the number of p16 positive cells was able to better discriminate the CIN lesions grades than the usual semi-quantitative analysis. The average Ki-67 labeling indexes for the normal epithelium, CIN I, CIN II, and CIN III groups were 19.8%, 27.3%, 32.8%, and 37.1%, respectively, whereas the mean MCM7 labeling indexes for the correspondent grades were 27.0%, 30.4%, 50.5%, and 67.2%. The Ki-67 and MCM7 labeling indexes were closely correlated with the CIN histological grade, with higher labeling indexe values obtained from the more severe lesions (p<0.05), being the MCM7 labeling indexes the highest values in all the CIN categories (p<0.05).

Conclusion

We observed a good correlation among the p16, Ki-67, and MCM7 data. In addition, MCM7 demonstrated to be a more efficient and sensitive marker to assess disease progression in the uterine cervix.     

The study of the combination detection of HPV-DNA and p16INK4a in cervical lesions

The objective of this study is to detect the infection of human papillomavirus (HPV) and the expression of p16(INK4a) in cervical lesions and to investigate the interaction between hrHPV and p16(INK4a) for cervical lesions and its diagnostic efficiency. hrHPV-DNA was detected by the hybrid capture II (HC-II) system. Immunochemical method was used to detect the expression of p16(INK4a), and histopathologic test was performed to identify cervical lesions. χ(2) test and Spearman's rank correlation were used for statistical analysis. Additive effects model was used to analyze the interaction. The diagnostic sensitivity, specificity, positive predictive values, negative predictive values, accuracy, and the area under the receiver operating characteristic curve were calculated with SPSS 13.0. hrHPV and p16(INK4a) positive rate increased (P < 0.05) with histopathologic diagnosis increasing. The positive rates of hrHPV and p16(INK4a) in negative or chronic inflammation were statistically lower than that in cervical intraepithelial neoplasia (CIN)1, CIN2, CIN3, and squamous-cell carcinoma (SCC) (P < 0.05), respectively. There was a positive interaction between hrHPV and p16(INK4a), relative excess risk of interaction (RERI) was 52.49, attributable proportions of interaction (API) were 72.34%, and the synergy index (S) was 3.75. The specificity and AUC of combining hrHPV with p16(INK4a) were statistically higher than hrHPV or p16(INK4a) alone (P < 0.05). hrHPV and p16(INK4a) are useful markers for the early diagnosis of cervical lesions. A positive interaction between hrHPV and p16(INK4a) is seen. The combination of hrHPV and p16(INK4a) has a higher diagnostic accuracy than hrHPV or p16(INK4a) alone in diagnosis of cervical lesions.     

p16INK4a免疫细胞化学检测在筛查宫颈癌中的作用

目的 探讨p16^INK4a免疫细胞化学检测在筛查宫颈癌及其癌前病变中的作用。方法 选择220例宫颈液基细胞学剩余标本,制作液基薄片进行p16^INK4a 免疫细胞化学检测,随访组织活检结果,并与高危人乳头瘤病毒（HR—HPV）DNA检测结果进行对照。结果 p16^INK4a在宫颈细胞学诊断的鳞状细胞癌（SCC）、鳞状上皮内高度病变（HSIL）、鳞状上皮内低度病变（LSIL）、非典型鳞状细胞-小除外上皮内高度病变（ASC—H）和非典型鳞状细胞-不能明确意义（ASC—US）病例的阳性表达率分别为100．0％（7／7）、92．2％（107／116）、24．3％（17／70）、100．0％（14／14）和36．4％（4／11）。150例p16^INK4a阳性者中,111例有组织活检诊断,其中宫颈上皮内瘤变（CIN）2级及以上病变者97例（87．4％）;70例p16^INK4a阴性者中,18例有组织活检诊断,无一例CIN2及以上病变。p16^INK4a在CIN2及以上病变与在CIN1之间的阳性表达率差异有统计学意义（P〈0.01）,而HR-HPV DNA的阳性率在两者之间差异无统计学意义。结论 p16^INK4a在宫颈HSIL及以上病变中高表达,有利于高危病例的筛选。     

Immunohistochemical analysis of estrogen receptors, progesterone receptors, Ki-67 antigen, and human papillomavirus DNA in normal and neoplastic epithelium of the uterine cervix.

To investigate the relationship between the sex steroid receptor (estrogen receptor [ER] and progesterone receptor [PR]) status and the cell proliferation kinetics during the menstrual cycle in normal and neoplastic epithelium of the uterine cervix, immunohistochemical localization of ER, PR, and cell proliferation-associated antigen, Ki-67, was investigated in 35 normal cervical specimens, 3 condylomas, 26 cervical intraepithelial neoplasia (CIN) samples, and 22 invasive squamous carcinoma samples. The presence of human papillomavirus (HPV) DNA was also studied. In the normal cervix, basal cells were usually ER positive, PR negative, and Ki-67 negative throughout the menstrual cycle. Parabasal cells were ER positive and PR negative in the follicular phase, but ER negative and PR positive, and Ki-67 positive in the luteal phase, and Ki-67-positive cells increased in number in the luteal phase. In contrast, PR positivity was observed in the cells of condyloma (2 of 2 cases), CIN (19 of 26 cases), and invasive squamous carcinoma (13 of 22 cases) irrespective of the menstrual phase. Moreover, most neoplastic cells containing HPV DNA type 16/18 were ER negative, whereas several lesions containing HPV DNA type 31/33/35 were weakly ER positive. Many Ki-67-labeled cells were observed in the neoplastic lesions. These results suggest that reduced ER expression and increased PR expression are associated with the proliferation of normal cervical squamous epithelium, and this proliferation-related receptor status, which is probably induced by HPV infection, is usually expressed in neoplastic cervical squamous cells.