宫颈细胞DNA倍体分析联合B7-H4和PKCδ对宫颈癌的诊断价值

摘要/Abstract

摘要：

目的探讨宫颈细胞DNA倍体分析联合负性共刺激分子B7同源物4（B7-H4）和蛋白激酶Cδ（PKCδ）对宫颈癌的诊断价值。方法选取2018年1月到2022年1月在石家庄市人民医院诊治的宫颈癌患者160例作为宫颈癌组，同期在本院做宫颈癌前筛查的女性160例作为对照组，根据检查结果分为正常或炎症组（n=52）、低级别宫颈上皮内瘤变（CIN）组（n=68）、高级别CIN组（n=40）。采用全自动细胞图像分析系统分析宫颈细胞DNA倍体情况。采用酶联免疫吸附法测定血清B7-H4、PKCδ水平。采用Pearson相关分析探讨血清B7-H4、PKCδ的相关性;采用受试者操作特征（ROC）曲线评估宫颈细胞DNA倍体分析联合血清B7-H4、PKCδ对宫颈癌的诊断价值;采用logistic多因素回归进行宫颈癌的危险因素分析。结果正常或炎症组、低级别CIN组、高级别CIN组、宫颈癌组DNA倍体阳性例数分别为16（30.8%）、27（39.7%）、26（65.0%）、127（79.4%），差异有统计学意义（H=55.86，P<0.001）。进一步两两比较发现，与正常或炎症组比较，高级别CIN组、宫颈癌组DNA倍体阳性比例均高（均P<0.05）;与低级别CIN组比较，宫颈癌组DNA倍体阳性比例高（P<0.05）。正常或炎症组、低级别CIN组、高级别CIN组、宫颈癌组血清B7-H4水平分别为（57.21±10.21）、（79.17±11.34）、（92.73±15.36）、（126.56±20.25）ng/ml，差异有统计学意义（F=285.45，P<0.001），血清PKCδ水平分别为（89.34±18.29）、（71.79±15.82）、（53.39±11.84）、（40.23±10.21）ng/ml，差异有统计学意义（F=216.28，P<0.001），进一步两两比较发现，正常或炎症组、低级别CIN组、高级别CIN组、宫颈癌组血清B7-H4水平依次升高（均P<0.05）;正常或炎症组、高级别CIN组、宫颈癌组血清PKCδ水平依次降低（均P<0.05）。Pearson检验分析结果显示，宫颈癌患者血清B7-H4、PKCδ水平呈负相关（r=-0.47，P<0.001）。ROC曲线分析显示，宫颈细胞DNA倍体诊断宫颈癌的曲线下面积（AUC）为0.82（95%CI为0.78~0.86），敏感性、特异性分别为83.9%、79.9%;血清B7-H4诊断宫颈癌的AUC为0.92（95%CI为0.89~0.95），敏感性、特异性分别为95.7%、76.1%，截断值为111.12 ng/ml;血清PKCδ诊断宫颈癌的AUC为0.92（95%CI为0.89~0.95），敏感性、特异性分别为85.6%、88.9%，截断值为54.83 ng/ml;三者联合诊断宫颈癌的AUC为0.99（95%CI为0.97~0.99），敏感性、特异性分别为98.3%、75.9%。三者联合诊断宫颈癌的AUC大于宫颈细胞DNA倍体（Z=8.00，P<0.001）、血清B7-H4（Z=4.34，P<0.001）、血清PKCδ（Z=4.61，P<0.001）单独诊断的AUC。多因素logistic回归分析结果显示，血清B7-H4高水平（OR=2.94，95%CI为1.78~4.84，P<0.001）、PKCδ低水平（OR=4.33，95%CI为1.88~10.00，P=0.001）、宫颈细胞DNA倍体阳性（OR=5.77，95%CI为2.38~13.99，P<0.001）均为影响宫颈癌发生的独立危险因素。结论宫颈癌患者宫颈细胞DNA倍体阳性比例升高，血清B7-H4水平升高、PKCδ水平降低，宫颈细胞DNA倍体分析联合血清B7-H4和PKCδ对宫颈癌具有较高的诊断价值。

关键词:宫颈肿瘤,蛋白激酶Cδ,早期诊断,宫颈细胞DNA倍体,负性共刺激分子B7同源物4

Abstract:

ObjectiveTo investigate the diagnostic value of cervical cell DNA ploidy analysis combined with negative costimulatory molecule B7 homolog 4 （B7-H4） and protein kinase Cδ （PKCδ） for cervical cancer.MethodsA total of 160 cervical cancer patients diagnosed and treated at Shijiazhuang People's Hospital from January 2018 to January 2022 were selected as the cervical cancer group. Meantime， 160 women who were screened for cervical cancer in our hospital during this period were selected as the control group. According to the examination results， they were divided into normal or inflammatory group （n=52）， low-grade cervical intraepithelial neoplasia （CIN） group （n=68） and high-grade CIN group （n=40）. The automatic cell image analysis system was used to analyze the DNA ploidy of cervical cells. The levels of B7-H4 and PKCδ in serum were determined by enzyme-linked immunosorbent assay. Pearson method was used to analyze the correlation between serum B7-H4 and PKCδ; the diagnostic value of cervical cell DNA ploidy analysis combined with serum B7-H4 and PKCδ in cervical cancer was evaluated by the receiver operator characteristic （ROC） curve; multivariate logistic regression was used to analyze the risk factors of cervical cancer.ResultsThe numbers of DNA ploidy positive cases of cervical cells in normal or inflammatory group， low-grade CIN group， high-grade CIN group and cervical cancer group were 16 （30.8%）， 27 （39.7%）， 26 （65.0%） and 127 （79.4%）， respectively， with a statistically significant difference （H=55.86，P<0.001）. Further pin-by-pair comparison showed that compared with normal or inflammatory groups， the proportion of DNA ploidy positive in high-grade CIN group and cervical cancer group were higher （bothP<0.05）. The proportion of DNA ploidy positive in cervical cancer group was higher than that in low-grade CIN group （P<0.05）. Serum B7-H4 levels in normal or inflammatory group， low-grade CIN group， high-grade CIN group and cervical cancer group were （57.21±10.21），（79.17±11.34），（92.73±15.36），（126.56±20.25） ng/ml， respectively， with a statistically significant difference （F=285.45，P<0.001）. Serum PKCδ levels were （89.34±18.29），（71.79±15.82），（53.39±11.84），（40.23±10.21） ng/ml， respectively， with a statistically significant difference （F=216.28，P<0.001）. Further pin-by-pair comparison showed that serum B7-H4 levels in normal or inflammatory groups， low-grade CIN group， high-grade CIN group and cervical cancer group increased in turn （allP<0.05）. Serum PKCδ levels in normal or inflammatory groups， high-grade CIN group and cervical cancer group were decreased in turn （allP<0.05）. Pearson correlation analysis showed that the serum B7-H4 and PKCδ levels in patients with cervical cancer were negatively correlated （r=-0.47，P<0.001）. ROC curve analysis showed that the area under the curve （AUC） of cervical cell DNA ploidy for cervical cancer diagnosis was 0.82 （95%CI： 0.78-0.86）， and the sensitivity and specificity were 83.9% and 79.9%， respectively. The AUC of serum B7-H4 in the diagnosis of cervical cancer was 0.92 （95%CI： 0.89-0.95）， the sensitivity and specificity were 95.7% and 76.1%， respectively， and the cutoff value was 111.12 ng/ml. The AUC of serum PKCδ for diagnosis of cervical cancer was 0.92 （95%CI： 0.89-0.95）， the sensitivity and specificity were 85.6% and 88.9%， respectively， and the cut-off value was 54.83 ng/ml. The AUC of the combined diagnosis of cervical cancer was 0.99 （95%CI： 0.97-0.99）， and the sensitivity and specificity were 98.3% and 75.9%， respectively. The AUC of the combined diagnosis of cervical cancer was higher than that of DNA ploidy （Z=8.00，P<0.001）， serum B7-H4 （Z=4.34，P<0.001）， and serum PKCδ （Z=4.61，P<0.001） alone. Multivariate logistic regression analysis showed that high level of B7-H4 in serum （OR=2.94，95%CI： 1.78-4.84，P<0.001）， low level of PKCδ （OR=4.33，95%CI： 1.88-10.00，P=0.001） and positive DNA ploidy in cervical cells （OR=5.77，95%CI： 2.38-13.99，P<0.001） were independent risk factors for cervical cancer.ConclusionThe positive proportion of DNA ploidy in cervical cells of patients with cervical cancer is increased， the serum B7-H4 level is increased， the PKCδ level is decreased， and cervical cell DNA ploidy analysis combined with serum B7-H4 and PKCδ has a high diagnostic value for cervical cancer.

Key words:Uterine cervical neoplasms,Protein kinase C-delta,Early diagnosis,Cervical cell DNA ploidy,Negative costimulatory molecule B7 homolog 4

张宁宁, 杨哲, 檀丽梅, 李振宁, 王迪, 魏永志. 宫颈细胞DNA倍体分析联合B7-H4和PKCδ对宫颈癌的诊断价值[J]. 国际肿瘤学杂志, 2024, 51(5): 286-291.

Zhang Ningning, Yang Zhe, Tan Limei, Li Zhenning, Wang Di, Wei Yongzhi. Diagnostic value of cervical cell DNA ploidy analysis combined with B7-H4 and PKCδ for cervical cancer[J]. Journal of International Oncology, 2024, 51(5): 286-291.

图/表5

表1

4组一般资料比较[$\bar{x}\pm s$/例（%）]"

指标	正常或炎症组（n=52）	低级别CIN组（n=68）	高级别CIN组（n=40）	宫颈癌组（n=160）	F/H值	P值
年龄（岁）	53.18±6.86	52.50±7.53	54.02±7.82	53.55±7.24	0.06	0.980
BMI（kg/m²）	25.91±3.38	24.79±3.07	24.12±2.84	25.12±3.11	0.46	0.713
绝经
是	11（21.2）	20（29.4）	9（22.5）	42（26.3）	1.30	0.730
否	41（78.8）	48（70.6）	31（77.5）	118（73.7）	1.30	0.730
吸烟
是	10（19.2）	8（11.8）	6（15.0）	16（10.0）	3.33	0.343
否	42（80.8）	60（88.2）	34（85.0）	144（90.0）	3.33	0.343
DNA倍体
阳性	16（30.8）	27（39.7）	26（65.0）^a	127（79.4）^ab	55.86	<0.001
阴性	36（69.2）	41（60.3）	14（35.0）	33（20.6）	55.86	<0.001

表1

组别	B7-H4	PKCδ
正常或炎症组（n=52）	57.21±10.21	89.34±18.29
低级别CIN组（n=68）	79.17±11.34^a	71.79±15.82
高级别CIN组（n=40）	92.73±15.36^ab	53.39±11.84^ab
宫颈癌组（n=160）	126.56±20.25^abc	40.23±10.21^abc
F值	285.45	216.28
P值	<0.001	<0.001

表2

图1

图2

变量	β值	SE值	Waldχ²值	OR值	95%CI	P值
B7-H4	1.08	0.26	17.85	2.94	1.78~4.84	<0.001
PKCδ	1.47	0.43	11.78	4.33	1.88~10.00	0.001
DNA倍体	1.75	0.45	15.03	5.77	2.38~13.99	<0.001

表3

参考文献26

[1]	Pála M. How unknown is metastatic carcinoma to cervical lymph nodes from an unknown primary?[J].Klin Onkol,2023,36(4): 364-369. DOI:10.48095/CCKO2023364. pmid:37877528
[2]	Shen SN, Zhang SB, Liu P, et al. Potential role of microRNAs in the treatment and diagnosis of cervical cancer[J].Cancer Genet,2020,248/249: 25-30. DOI:10.1016/j.cancergen.2020.09.003.
[3]	Steely AM, Watkins AA, Vidal B, et al. Pembrolizumab improves survival after resection of intracardiac metastatic cervical carcinoma[J].Ann Thorac Surg,2022,114(5): e323-e325. DOI:10.1016/j.athoracsur.2021.12.066.
[4]	Tan ZB, Shen WX. Prognostic role of B7-H4 in patients with non-small cell lung cancer: a meta-analysis[J].Oncotarget,2017,8(16): 27137-27144. DOI:10.18632/oncotarget.15648. pmid:28404927
[5]	郭庆枝, 李丽, 薄其美, 等. 宫颈癌组织中PKCα、PKCε、PKCδ表达及与化疗药物敏感性的机制研究[J].海南医学院学报,2019,25(2): 120-124. DOI:10.13210/j.cnki.jhmu.20181217.002.
[6]	中国抗癌协会妇科肿瘤专业委员会. 宫颈癌诊断与治疗指南(第四版)[J].中国实用妇科与产科杂志,2018,34(6): 613-622. DOI:10.19538/j.fk2018060111.
[7]	Buskwofie A, David-West G, Clare CA. A review of cervical cancer: incidence and disparities[J].J Natl Med Assoc,2020,112(2): 229-232. DOI:10.1016/j.jnma.2020.03.002. pmid:32278478
[8]	Zhang YY, Wang JL, Zhang R, et al. Application value of detection of high-risk HPV infection in early cervical cancer patients in disease diagnosis and prognosis evaluation[J].Clin Lab,2020,66(12). DOI:10.7754/Clin.Lab.2020.200245.
[9]	Sankaranarayanan R, Joshi S, Muwonge R, et al. Can a single dose of human papillomavirus (HPV) vaccine prevent cervical cancer? Early findings from an Indian study[J].Vaccine,2018,36(32 Pt A): 4783-4791. DOI:10.1016/j.vaccine.2018.02.087. pmid:29551226
[10]	Roca AM, Chobrutskiy BI, Callahan BM, et al. T-cell receptor V and J usage paired with specific HLA alleles associates with distinct cervical cancer survival rates[J].Hum Immunol,2019,80(4): 237-242. DOI:10.1016/j.humimm.2019.01.005. pmid:30684568
[11]	Baral G. Cervical cancer screening and HPV vaccination in Nepal[J].South Asian J Cancer,2023,12(1): 53-54. DOI:10.1055/s-0043-1764155. pmid:36851932
[12]	Nassiri S, Aminimoghaddam S, Sadaghian MR, et al. Evaluation of the diagnostic accuracy of the cervical biopsy under colposcopic vision[J].Eur J Transl Myol,2022,32(4): 10670. DOI:10.4081/ ejtm.2022.10670.
[13]	翟瑞芳, 郭嘉婧, 徐晓燕, 等. DNA倍体高危型HPV ki67 p16在宫颈鳞癌患者转移阴性盆腔淋巴结中的检测及意义[J].中国药物与临床,2019,19(13): 2194-2196. DOI:10.11655/zgywylc2019.13.021.
[14]	Cang W, Li Q, Gu LY, et al. Clinical evaluation of DNA ploidy for the triage of HPV-positive Chinese women during cervical cancer screening[J].Cancer Prev Res (Phila),2021,14(3): 355-362. DOI:10.1158/1940-6207.CAPR-20-0229.
[15]	Guo YL, Peng QZ, Wang Y, et al. The application of DNA ploidy analysis in large-scale population screening for cervical cancer[J].Acta Cytol,2021,65(5): 385-392. DOI:10.1159/000518052. pmid:34482310
[16]	贺丹丹, 毛渭东, 李志龙. 细胞DNA定量分析在宫颈癌及癌前病变早期筛查中应用的临床观察[J].山西医药杂志,2019,48(18): 2285-2286. DOI:10.3969/j.issn.0253-9926.2019.18.032.
[17]	胡晓君, 雷磊, 胡艳, 等. B7-H4在宫颈癌组织中的表达及与临床病理特征的关系[J].现代肿瘤医学,2021,29(6): 1012-1016. DOI:10.3969/j.issn.1672-4992.2021.06.023.
[18]	Wang JY, Wang WP. B7-H4, a promising target for immunotherapy[J].Cell Immunol,2020,347: 104008. DOI:10.1016/j.cellimm.2019.104008.
[19]	Shen LW, Qian Y, Wu WG, et al. B7-H4 is a prognostic biomarker for poor survival in patients with pancreatic cancer[J].Hum Pathol,2017,66: 79-85. DOI:10.1016/j.humpath.2017.05.023. pmid:28600225
[20]	MacGregor HL, Garcia-Batres C, Sayad A, et al. Tumor cell expression of B7-H4 correlates with higher frequencies of tumor-infiltrating APCs and higher CXCL17 expression in human epithelial ovarian cancer[J].Oncoimmunology,2019,8(12): e1665460. DOI:10.1080/2162402X.2019.1665460.
[21]	Blasiak J. DNA-damaging anticancer drugs—a perspective for DNA repair-oriented therapy[J].Curr Med Chem,2017,24(15): 1488-1503. DOI:10.2174/0929867324666170124145557. pmid:28120709
[22]	Mohanty S, Huang J, Basu A. Enhancement of cisplatin sensitivity of cisplatin-resistant human cervical carcinoma cells by bryostatin 1[J].Clin Cancer Res,2005,11(18): 6730-6737. DOI:10.1158/1078-0432.CCR-05-0450. pmid:16166454
[23]	Zhou B, Lu YW, Zhao ZM, et al. B7-H4 expression is upregulated by PKCδ activation and contributes to PKCδ-induced cell motility in colorectal cancer[J].Cancer Cell Int,2022,22(1): 147. DOI:10.1186/s12935-022-02567-1.
[24]	周艳, 陈晓西. 宫颈细胞DNA倍体定量分析联合液基薄层细胞学检查在宫颈癌早期筛查中的应用价值[J].癌症进展,2022,20(12): 1239-1241. DOI:10.11877/j.issn.1672-1535.2022.20.12.13.
[25]	潘银燕, 李世宏, 宋娟, 等. 液基细胞学、HPV检测及DNA倍体定量分析在宫颈癌筛查中的价值[J].中国妇产科临床杂志,2023,24(1): 69-70. DOI:10.13390/j.issn.1672-1861.2023.01.021.
[26]	刘畅, 李坤, 舒丽莎, 等. B7-H4和Foxp3⁺Treg在宫颈病变组织中的表达及临床意义[J].重庆医学,2019,48(17): 2960-2963. DOI:10.3969/j.issn.1671-8348.2019.17.019.