The Expression Of B7-H4 In Tumor Tissues And The Diagnosis Values Of B7-H4, HE4, CA125 Assay In Serum Of Patients With Epithelial Ovarian Cancer

Objective: Ovarian cancer is a malignant tumor with the highest mortality in gynecological tumors. Its pathogenesis is hidden, but developed rapidly, about 70%～80%of ovarian cancer patients are at advanced stage when they are diagnosed, the 5-year survival rate is about 20%～30%, the main cause is that the patient with early ovarian cancer has no symptoms, and it is difficult to see a doctor at early stage. So how to diagnose ovarian cancer early is becoming the urgent problem, which needs to be solved. This study is to detect the expression of B7-H4 (B7X) in tissiues of normal ovaries, ovarian benign tumors, borderline ovarian tumors and ovarian cancer; the serum expression of B7-H4 and Human Epididymis protein 4 (HE4). And explored the correlation of them with ovarian cancer. Exploring the ability of B7-H4 and HE4 as tumor markers combined with CA125 for early diagnosis, monitoring curative effect.Methods: 1 Immunohistochemistry was used to detect the expression of B7-H4 protein in normal ovaries and epithelial ovarian tumors, among these series, 11 cases were normal ovaries, 20 cases were benign ovarian tumors, 10 cases were borderline ovarian tumors and 30 cases were ovarian cancer . Including serous cystadenocarcinoma(n=25), mucinou cystade- nocarcinoma (n=5). The correlation between positive expression rate of epithelial ovarian cancer tissues and clinicopathological characters were analyzed.2 The double antibody sandwich Enzyme-linked Immu- nosorbent Assay (ELISA) technique was applied to detect the serum concentration of B7-H4 and HE4. Among the series, 11 healthy women, 20 benign ovarian tumors, 7 borderline ovarian tumors, preoperative and postoperative serum of 25 ovarian cancer patient. CA125 concentration was detected in our laboratory by an electrogenerated chemilumincence method. The expression levels of B7-H4 and HE4 in four groups were compaired, respectively. The correlation between preoperative serum concetration of B7-H4, HE4 and different clinico- pathological characters were anylazed. The preoperative and postoperative serum concetration of B7-H4 and HE4 in cancer group were anylazed, respectively. The correlation between serum concetration of B7-H4 and tissue expression intensity was anylazed.3 Made operating characeristic curves(ROC) by using healthy and benign tumor groups as control and got the cut-off point. The sensitivity and specificity of B7-H4 and HE4 were calculated. The ability of serum HE4 at different cut-off point in diagonosing epithelial ovarian cancer was analyzed.4 All statistical analyses were performed with SAS statistical software package, measurement data was expressed with median, the comparisons of multiple groups were analyzed by Kruskal-wallis test, and the two-two comparisons among groups were done by Bonferronit t test. Wilcoxon signed rank test was used for the anlysis of paired data. The differences of the enumeration data were compared with the chi-square test. The positive expression rate in different pathological types, the clinical stages and pathological grades were compared with the chi-square test and the Fisher's exact probability test. The correlation between serum concetration of and tissue expressin intensity was evaluated by spearman correlation anylasis. The level of significance was setted at P<0.05.Results: 1 In normal ovaries, benign ovarian tumors, borderline ovarian tumors and ovarian cancer, the positive expession rates of B7-H4 protein were 9.1%(1/11), 20%(4/20), 30%(3/10) and 83.3%(25/30), respectively. The differences of positive expression rates in the four groups were significant (χ2=29.2901, P<0.0001). The level of last group was higher than groups before (P<0.0071). No significantly statistical difference was found between other two-two comparisons (P>0.0071).2 The correlation between tissue expression of B7-H4 and different clinicopathological charactersThe positive expression rate of B7-H4 protein in serous cystadenocarcinoma 92%(23/25) was higher than that of mucinous cystadenocarcinoma 40%(2/5), (P=0.0219); The positive expression rate was lower in ovarian cancer patients with early stages (stageⅠ/Ⅱ, 57.1%) compared with the advanced stages (stageⅢ/Ⅳ, 95.7%), (P=0.0307<0.05);the positive rate in well differentiation group (62.5%) was lower than moderately and poorly differentiation groups (95.5%) (P=0.0475).3 The serum concetration of B7-H4 and HE4 in healthy women, benign ovarian tumors, borderline ovarian tumors and ovarian cancer groupsThe serum concentrations of B7-H4 and HE4 protein in cancer group were higher than groups former, (both P<0.05), no difference was found between other groups, (all P>0.05). The median serum level of CA125 was 324.60(U/L). The serum levels of B7-H4 and HE4 on 3 days after operation were lower than before operation (P<0.0001).4 The correlation between serum concetration of B7-H4, HE4 and different clinicopathological characters The expression of B7-H4 was correlated with stage, grade, type, (all P<0.05); was not correlated with age and CA125, (both P<0.05). The expression of HE4 was correlated with type, age and CA125,(all P<0.05); was not correlated with stage and grade, (both P<0.05).5 Using the healthy women and benign tumor group as control, the diagnosis efficiency were the best when the B7-H4 and HE4 were at the cut-off point, which determined from ROC curve, 0.78ng/ml and 107.15pmol/L, respectively. The operating characteristic-area under the curve (ROC-AUC) of B7-H4 was 0.865, sensitivity was 84%, specificity was 78.9%. The ROC- AUC of HE4 was 0.960, sensitivity was 88%, and specificity was 92.1%.6 When the cut-off point of CA125 was 35KU/L, B7-H4 was 0.78ng/ml, HE4 was 107.15pmol/L(received from ROC) or 150pmol/L(received from agent kit): there was no difference for ovarian cancer detection by using B7-H4, HE4, CA125 alone, (P>0.05). B7-H4 or HE4 combined with CA125, and using three of them, the sensitivity in detecting for ovarian cancer were higher than CA125 alone, (P<0.05). The sensitivity of HE4 for detecting early ovarian cancer (stage I/II), which included 5 cases in this study, was the highest 100%, but B7-H4 and CA125 was 80%, 60%, respectively.7 There was no statistical difference in sensitivity and specificity of HE4 when the cut-off was 107.15pmol/L or 150 pmol/L (P>0.05).Conclusion: 1 The B7-H4 protein in ovarian cancer tissue was over expressed. And it was positively correlated with adverse clinicopathological stages, grades, and types. The serum levels of B7-H4 could represent the tissue expression, so B7-H4 protein may participate the cancer genesis and progression.2 B7-H4 and HE4 were expected to become the screening marker for ovarian cancer. B7-H4 expression level in serum of ovarian cancer had positive correlation with adverse clinicopathological stages, grades, and types. B7-H4 expression level was not correlated with age and CA125. HE4 expression level had positive correlation with types, ages and CA125. But it was not correlated with stages and grades. B7-H4 and HE4 could be complementary to CA125 for ovarian cancer diagnosis.3 There was no difference for ovarian cancer detection by using B7-H4, HE4, CA125 alone, (P>0.05). B7-H4 or HE4 combined with CA125, and using three of them, the sensitivity in detecting for ovarian cancer were higher than CA125 alone, (P<0.05). B7-H4 and HE4 were expected to become the new indexes for monitoring surgery effect and postoperative condition.4 When the cut-off point of HE4 was 107.15pmol/L, it was superior to 150p mol/L for improving the diagnostic sensitivity, accuracy, reducing the misdiagnostic rate. While the cut-off point was 150pmol/L, it was superior to 107.15pmol/L for decreasing the omission diagnostic rate.