Open Med. 2016; 11:125-132 Research Article Open Access Li-e Zheng*, Jun-ying Qu, Fei He The diagnosis and pathological value of combined detection of HE4 and CA125 for patients with ovarian cancer DOI 10.1515/med-2016-0024 other. Conclusion HE4 can be used as a novel clinical bio- received January 28, 2016; accepted March 9, 2016 marker for predicting malignant ovarian tumors and its expression was closely related with the clinical pathologi- Abstract: Objective To evaluate the value of individual and cal features of malignant ovarian tumors. combined measurement of human epididymis protein 4 (HE4) and cancer antigen 125 (CA-125) in the diagnosis Keywords: HE4; CA125; diagnosis; ovarian tumor of ovarian cancer. Methods A clinical case-control study was performed in which the levels of serum HE4 and CA-125 of subjects with malignant, borderline, benign ovarian tumors and healthy women were measured before 1 Introduction surgery. An immunohistochemistry method was used In the female reproductive system, ovarian cancer ranks to measure the expression of HE4 in different tissues. third in incidence and 1st in mortality in gynecological Statistical analysis was performed to determine the rela- malignancies [1]. Nearly 70% of patients with ovarian tionship between the level of HE4 and the pathologic cancer are diagnosed at an advanced stage, and the 5 year type as well as the stage of the ovarian tumors. Results survival rate for patients with ovarian cancer rises up to The level of HE4 in the serum was significantly elevated in 90% through early effective treatment [2]. Hence, diag- the malignant ovarian cancer group compared with other nosing at early stage is crucial to improve the prognosis of groups. Women with benign ovarian tumors and non-neo- ovarian cancer. Currently, the most commonly used serum plastic lesions, and healthy women were designated as biomarkers for early diagnosis of ovarian cancer is CA125. references. When the level of HE4 in the serum was 58.66 But it could be also elevated in serum for patients with pmol/L, the sensitivity and specificity of HE4 in diagnos- pelvic inflammatory disease, such as endometriosis peri- ing malignant ovarian tumors was 82.35% and 96.03%, toneal disease and injury [3]. So the application of CA125 in respectively. The level of HE4 was negatively correlated ovarian cancer diagnosis showed high false positive rate, with the differentiation extent of the tumors whereas pos- and sometimes this would lead to unnecessary surgery. itively correlated to the clinical staging. In the groups of Human epididymis protein 4 (Human epididymis gene malignant and borderline tumors, the levels of HE4 were product 4, HE4) is considered a new biomarker for early higher than the other groups. The expression of HE4 was diagnosis of ovarian cancer. In this study, we included a significant higher in the serous types of ovarian tumors case-control study to determine the serum level as well as than that of the mucous types (P<0.05). The level of HE4 in the expression of HE4 and CA125 in clinical ovarian cancer the serum and tissues were positively correlated with each tissues. We also intended to evaluate the value of individ- ual and/or combined measurement of HE4 and CA-125 in the diagnosis of ovarian cancer and prognosis prediction. *Corresponding author: Li-e Zheng, The First Affliliated Hospital Of Fujian Medical University, Chazhong Road 22, Fuzhou, Fujian 350001, People’s Republic of China, Tel:0591-87982062, E-mail: [email protected] Fujian Medical University 350004, China Jun-ying Qu, Fei He, Department of obstetrics and gynecology, the First Affliliated Hospital Of Fujian Medical University, Fujian Fuzhou 350005, China; Fujian Medical University 350004 ,China © 2016 Li-e Zheng et al published by De Gruyter Open This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License. 126 Li-e Zheng et al 2 Material and method age of malignant ovarian cancer group was from 11 to 74, with an average age of 49.43±14.69 years old. Non-ovarian cancer group was aged from 15 to 76, with an average age 2.1 Patients of 40.66±12.56. The patients were not receiving any other treatment in our hospital. Over the same period, we also Select the patients with a pelvic mass, who were first tested the serum level of HE4 and CA125 in healthy women inpatient for gynecological surgery in the First Affiliated (59 cases). All the subjects of our study were in non-men- Hospital of Fujian Medical University, from December 2010 strual period, with no breast/thyroid disease, and a to October 2012. All patients had no significant syndrome history of other cancers. that effecting tumor biomarkers, and received no preop- erative chemotherapy and other treatments. According Ethical approval: The research related to human use to the postoperative pathology judging results, the sub- has been complied with all the relevant national regula- jects were divided into malignant ovarian tumor group (58 tions, institutional policies and in accordance the tenets cases), borderline ovarian tumor group (10 cases), benign of the Helsinki Declaration, and has been approved by ovarian tumors (40 cases), ovarian non-neoplastic lesions the authors’ institutional review board or equivalent (27 cases) and other gynecologic malignancies (26 cases). committee. Among the 58 cases of malignant ovarian tumors, there were 42 cases of epithelial ovarian tumors and 16 cases of Informed consent: Informed consent has been obtained non-epithelial ovarian tumors. In addition, the malignant from all individuals included in this study. ovarian tumor group included 30 cases of serous carcino- mas, 12 cases of mucous carcinomas, 6 cases of ovarian endometrioid adenocarcinomas, 3 cases of clear-cell car- 2.2 Method cinomas , 4 cases of yolk sac tumors, and 3 cases of dys- germinoma. Clinical staging was classified according to The electrochemical luminescence method was used the International Federation of Gynecology and Obstetrics to detect the level of HE4 and CA125 in the serum. We (FIGO) criteria: 18 cases of I, 6 cases of II, 30 cases of III, applied the human epididymis protein 4 assay kit (Roche, and 4 cases of IV stage. There were 8 cases of highly dif- Shanghai, China, 05950929190),Roche cobas e 601 and ferentiated, 18 cases of moderately differentiated and 32 electrochemiluminescence assay. cases of poorly differentiated. There were 22 patients with Monoclonal mouse anti-human HE4 (ZM- viscera metastasis. The borderline ovarian tumor group 0413) and CA125 (ZM-0019) antibodies were ordered included 6 cases of borderline serous papillary tumors, from Beijing Zhongshan Golden Bridge Company. 4 cases of borderline mucous cystadenomas. The benign Immunohistochemical detection reagents (PV-9000) were ovarian tumor group included 17 cases of serous cysta- also purchased from Beijing Zhongshan Golden Bridge denomas, 12 cases of mucous cystadenomas, 5 cases of Company. Take the above paraffin blocks of ovarian mature teratomas, 4 cases of simple cysts, and 2 cases cancer tissues, and perform routine HE staining, and then of fibromas. Ovarian non-neoplastic lesions included observe the morphology of tumor cells through the micro- 27 cases of ovarian endometriosis cysts. Gynecological scope. Phosphate buffer was used as a negative control cancer group included 2 cases of uterine sarcomas, 6 cases instead of primary antibody. of endometrial cancer and 18 cases of cervical cancer. All The reference value of serum HE4 is 0-140pmol/L, blood samples were taken from the elbow in the day before regarding HE4>140pmol/L as positive. The reference value surgery, fasting venous whole blood 3 ml, placed in a test of serum CAl25 is 0~35U/ml, regarding CAl25>35U/ml as tube without anticoagulant, Specimen collection was positive. Serum HE4 diagnostic cutoff points was obtained stand at room temperature for 1 hour, centrifuged (3000 through the ROC curve. Ovarian tumors were used as r/min) to take the clear phase, and stored in -80 degrees positive controls, with PBS instead of primary antibody C freezer for testing. After reading the pathological sec- as blank control. Comprehensive observing each slice, tions of these patients, select respective paraffin blocks regard the brown particles appearing in cytoplasm as pos- for section at the thickness of 4 micrometers. Meanwhile, itive. Two pathologists read the piece randomly. The deter- we selected 10 cases of partial ovarian normal tissues mine criteria was shown as follows: staining were divided from the surgically hysteromyoma resections followed into four levels: almost not colored for 0 point, yellow for by 4% paraformaldehyde-fixed, paraffin-embedded, and 1 point, brown for 2 point, tan for 3 point; coloring cells in histologically confirmed by conventional HE staining. The percentage: ≤ 5% as 0, 6% -25% as 1, 26-50% as 2, ≥ 51% as HE4 and CA125 for patients with ovarian cancer 127 3. The degree of ovarian tissue coloring, coloring percent- 4.2 Serum HE4 and CA125 joint evaluation age score: 0-1 as negative, 2-3 as +, 4-6 as ++, 6 points or more as +++. In the group of malignant ovarian cancer, including serous ovarian cancer, mucous ovarian cancer and ovarian endometrium adenoendometriocarcinoma, there was about 66.67% of those CA125 negative cases emerged 3 Statistical analysis as HE4 positive. In the group of non-neoplastic lesions of ovarian, there was about 88.00% of those CA125 positive Statistical analysis were performed using SPSS 13.0. The cases emerged as HE4 negative. level of HE4 and CA125 were expressed as median number The relationship between the serum HE4 level and (M). The pathologic diagnosis was considered as the stan- the patient age, pathological type, grading and clini- dard, the ROC curve was traced, and the sensitivity and cal staging in the malignant ovarian tumor group before specificity were calculated.