The Expression And Significance Of KLK6,KLK8 Of Serum And Tumor Tissue In Patients With Epithelial Ovarian Cancer

Objective: Ovarian carcinoma is a malignant tumor with the highest mortality in gynecological tumor. Its pathogenesis is hidden, but developed rapidly, about 70%～80%of ovarian carcinoma are at advanced stage when they were diagnosed, the 5-year survival is about 20%～30%, the main cause is that the patient with early ovarian carcinoma has no symptoms, and it is difficult to see a doctor at an early stage, so, how to diagnose ovarian carcinoma early is becomeing the urgent problem, which is to be solved. This study is to detect the expressions of Kallikrein Gene 6(KLK6) and Kallikrein Gene 8(KLK8) protein in serum and tissiue of epithelial ovarian benign tumor, borderline tumor and ovarian carcinoma, and to explore the role of them in the development and invasion of ovarian carcinoma. Compared with CA125, to evaluate them for early diagnosis and follow up, to provide an experimental basis for therapy of ovarian carcinoma.Methods: 1 Immunohistochemistry was used to detect the expres- sions of KLK6, KLK8 protein in 93 epithelial ovarian tumor patients, among this series, 20 cases were benign ovarian tumor, 11 cases were borderline ovarian tumor and 62 cases were ovarian carcinoma. Including serous cystadenocarcinoma (n =33), mucinous cystadenocarcinoma (n=18) and endometri- oid cystadenocarcinoma (n=11). Mean gray degree of the positive cell of staining section(A value) was measured by computer image processing software.2 The positive expression rate of KLK6,KLK8 protein in benign ovarian tumor, borderline ovarian tumor and ovarian carcinoma was compared, relationship between expression of KLK6, KLK8 protein and different clinicopathological charact- er in epithelial ovarian cancer tissue was analyzed.3 The double antibody sandwich Enzyme-linked Immu- nosorbent Assay(ELISA) technique was applied to detect serum human Kallikrein 6(hK6) protein and human Kallikrein 8 (hK8) protein concentration, CA125 concentration from our laboratory was detected by an electrogenerated chemilumin- cence method.4 The levels of hK6, hK8, CA125 in benign ovarian tumor, borderline ovarian tumors and ovarian carcinoma were comp- ared. The correlation between serum concentrati on and tissue A value of KLK6 and KLK8 protein in epithelial ovarian cancer patients were analyzed.5 According to the positive standards from references published, sensitivity and specificity of serum hK6, hK8, CA125 in diagnosing ovarian carcinoma were compared.6 All statistical analyses were performed with spss12.0 statistical software package, measurement data was expressed as mean SD values, the comparison of multiple mean with analysis of variance, after the equal check of variance, and the two-two comparison among the means was done by LSD-t method. The differences of the enumeration data was compared with the chi-square test. The positive expression rate in different pathological types, the clinical staging and pathological grade was compared with the chi-square test and the fisher's exact probability test. The correlation between serum concentration and tissue A value was evaluated by spearman correlation analysis. The level of significance was set at P<0.05.Results: 1 In benign ovarian tumor, borderline ovarian tumor and ovarian carcinoma, positive expression rate of KLK6 protein was 15%, 63.6%, 61.3%,respectively. The positive expression rate of benign ovarian tumor was significantly lower than those of the last two groups (P<0.01). The last two groups were further compared, no statistically significant difference (P> 0.05). In serous cystadenocarcinoma, mucinous cystaden ocarcinoma and endometrioid adenocarcin- oma, The positive expre- ssion rate of KLK6 protein was 78.8% (26/33), 44.4% (8/18), 36.4%(4/11), respectively. The positive expression rate of serous cystadenocarcinoma was signficantly higher than those of the last two groups (P<0.05), the last two groups were further compared, no statistically significant difference (P> 0.05); The positive expression was significantly lower in ovari- an carcinoma patients with the early stages (stageⅠ,Ⅱ, 41.7%) compared with the advanced stages (stageⅢ,Ⅳ, 73.1%) (P< 0.05);the positive rate in well differentiation group (40%) was lower than that in moderately and poorly differentiation group (71.4%) (P<0.05).2 In benign ovarian tumor, borderline ovarian tumor and ovarian carcinoma, the positive expression rate of KLK8 protein was 25%, 27.3%, 66.1%, respectively. The positive expression rate of ovarian carcinoma group was signficantly higher than those of the former two groups (P<0.05). The former two groups were further compared, no statistically significant difference (P>0.05). In serous cystadenocarcinoma, mucinous cystadenocarcinoma and endometrioid adenocarcino- ma, the positive expression rate of KLK8 protein was 87.9%, 50%, 27.3%, respectively. The positive expression rate of serous cystadenocarcinoma was signficantly higher than those of the last two groups (P<0.05). The last two groups were further compared, no statistically significant difference (P> 0.05); Significantly higher in ovarian carcinoma patients with the early stages (stageⅠ,Ⅱ, 83.3%) compared to the advanced stages (stageⅢ,Ⅳ, 55.3%) (P<0.05);The positive rate in well differentation group (85%) was higher than that in moderately and poorly differentiation group(57.1%) (P<0.05).3 In benign ovarian tumor, borderline ovarian tumor and ovarian carcinoma, the expression level of serum hk6 protein was 3.24±0.19, 3.88±0.22, 4.49±0.19(μg/L), respectively; hk8 protein was 4.26±0.29, 5.26±0.46, 6.59±0.15(μg/L), CA125 was 18.58±16.45, 36.22±15.24, 146.78±110.62 , respectively. The expression levels of serum hK6, hK8, CA125 were difference among three groups (P<0.01 or P< 0.05),hK6 protein concentration in benign ovarian tumor was lower than those in the last groups, the last two groups were further compared, in the ovarian carcinoma group, it was significantly higher than that in the borderline ovarian tumor group (P<0.01)。HK8 protein concentration in ovarian carcinoma was higher than those in the former groups, the former two groups were further compared, no statistically significant difference (P>0.05). CA125 concentration in benign ovarian tumor was lower than those in the last two groups (P<0.05), the last two groups were further compared, the levels of expression in ovarian carcinoma group was significantly higher than that in the borderline ovarian tumor group (P<0.05).4 The A value of KLK6, KLK8 protein in ovarian carcino- ma tissue was 145.2±11.3, 156.4±14.7,respectively, which we- re positively correlated with serum concentration, their correl- ation coefficients was 0.31(P<0.05), 0.608(P< 0.001).5 According to the positive standards from references published, sensitivity and specificity of serum hK6, hK8, CA125 in diagnosing ovarian carcinoma was 66.1%, 61.3%, 69.4%; 83.9%, 77.4%, 69.4%, respectively, among three markers, no statistically significant difference(P>0.05).Conclusions: 1 The expression of KLK6, KLK8 proteins are up-regulati on in serum and tissue of patients with epithelial ovarian carcinoma. The changes of their expressions are related to clinical pathology stage and malignant degree of ovarian carci- noma, KLK6, KLK8 cou1d be seen as an marker for the risk classification of ovarian carcinoma. Up-regulation of KLK6 expression indicates poor prognosis, but for KLK8, indicates well prognosis.2 The levels of serum hk6 and hk8 protein are related with the levels of KLK6 and KLK8 protein in epithelial ovarian carcinoma.3 KLK6, KLK8 are involved in the development of epith- elial ovarian carcinoma, combined serum tumor markers (hK6, hK8 and CA125) can be used to guide in early diagnosis, metastases and pathological staging and chemiotherapy of ovarian carcinoma.4 The particularity of KLK6, KLK8 protein in structure, biodistribution and metabolism, which makes them become the new serum marker of ovarian carcinoma, and they could be considered as possible targets for the gene therapy of ovarian carcinoma.