The Expression Of Protein BRCA1、HMGB1 And P62 In Epithelial Ovarian Carcinoma And Their Clinical Significance

Objective:To study the resistant mechanism of ovarian cancer in vitro and to investigate the expression of autophagy related gene HMGB1, P62 in epithelial ovarian cancer (EOC). To detect the protein expression of BRCA1, HMGB1 and P62 in the epithelial ovarian carcinoma by immunohistochemical method SP. The relationship between the two proteins and the histology degree, the criterion of FIGO, chemotherapy resistance and the relevance of the two types of protein and to evaluate the role of BRCA1, HMGB1 and P62 on the origin、progression、therapy and prognosis of epithelial ovarian cancer; and to illustrate the relationship between BRCA1, HMGB1 and P62 act as the dependence of choosing clinic therapy, predicting prognosis and finding effective retro-drug-resistance methods.Methods (1) CCK-8 was used to detect the influence of varied doses and time of cisplatin on the growth of SKOV3 and SKOV3/DDP cells, and the use of RT-PCR, Western Blotting to detect the expression of HMGB1 and P62. (2) Immunohistochemistry method was used to detect the expression of BRCA1 HMGB1 and P62 protein in 38 epithelial ovarian carcinoma,13 benign epithelial ovarian tumor tissues and 8 borderline ovarian tumors tissue samples. Among 39 epithelial ovarian carcinoma were 23 serous cystadenocarcinoma,4 mucinous cystadenocarcinoma,5 endometrioid carcinoma,5 clear cell carcinoma,2 mixed carcinoma; they were graded according to histology degree:well-differentiated ovarain carcinoma,8samples,moderated and poorly differentiated ovarain carcinoma, 31 samples; Clinical stage according the criterion of FIGO, Stage Ⅰ～Ⅱ,19 samples, Stage Ⅲ-Ⅳ,20 samples; Platinum sensitive 26 cases,13 cases of platinum Resistance.(3) Using the SPSS 19.0 to process the data, Chi-square test and spearman correlation analysis to statistically analyze the data. Statistical results of P<0.05 was considered statistically significant.Results1.DDP could inhibit the growth of SKOV3 and SKOV3/DDP cells in a time-and dose-dependent fashion. The study found that HMGB1 mRNA and protein expression in SKOV3 cells than SKOV3/DDP cells. P62 mRNA and protein expression in SKOV3 cells than SKOV3/DDP cells.2. Immunohistochemistry results show that BRCA1 protein was located in the Ovarian nuclei, Which was brown granules. The P62 protein was located in the ovarian cytoplasm, Which was brown granules.The HMGB1 protein was located in the ovarian nuclei and partial cytoplasm, Which was brown granules3. Expression of BRCA1:For the expression of BRCAl, there are 7 in of 13 benign epithelial ovarian tumor tissues samples show positive(53.8%); 4 of the 8 cases appear to be positive in the borderline ovarian tumors tissue (50%); 8 are seen to be positive in the 39 cases of the epithelial ovarian carcinoma(23.1%). The positive expression rate of BRCA1 protein in the epithelial ovarian carcinoma was showed lower than that in benign ovarian tumor and borderline ovarian tumors tissue, Which was significantly different (x2=6.440, P=0.040). The protein expression of BRCA1 was not correlated to clinical stage(P>0.05). In epithelial ovarian cancer BRCA1 protein in platinum sensitive positive expression is lower than drug-resistant ovarian cancer, with a difference (x2=7.863, P=0.005).4. Expression of HMGB1:For the expression of HMGB1, there are 12 in of 13 benign epithelial ovarian tumor tissues samples show positive(92.3%); 8 of the 8 cases appear to be positive in the borderline ovarian tumors tissue (100%); 38 are seen to be positive in the 39 cases of the epithelial ovarian carcinoma(97.4%). The positive expression rate of HMGB1 protein in the different tumors tissue was not significantly (P>0.05). the expression of HMGB 1 protein was correlated tobhistological type, and they were no significant correlation with the age, grade(P>0.05). In epithelial ovarian cancer HMGB1 protein in platinum sensitive and drug-resistant ovarian cancer were not significantly (P>0.05).5. Expression of P62:For the expression of P62, there are 0 in of 13 benign epithelial ovarian tumor tissues samples show positive(0%); 4 of the 8 cases appear to be positive in the borderline ovarian tumors tissue (50%);21 are seen to be positive in the 39 cases of the epithelial ovarian carcinoma(53.8%). The positive expression rate of P62 protein in the epithelial ovarian carcinoma was showed higher than that in benign ovarian tumor and borderline ovarian tumors tissue, Which was significantly different (X2=11.117, P=0.004). The protein expression of P62 was correlated to clinical stage, and they were no significant correlation with the age, grade and histological type. In epithelial ovarian cancer P62 protein in platinum sensitive positive expression is higher than drug-resistant ovarian cancer, with a difference (x2=6.174, P=0.013).6. There was significant positive correlation between HMGB1 and P62 protein expression in human ovarian cancer (r=-0.399, P=0.012).Conclusions1. HMGB1 and P62 mRNA and protein expression in SKOV3/DDP rate cut, hints of HMGB1 and P62 expression in ovarian cancer platinum resistance plays an important role, may be related to the occurrence and development of epithelial ovarian sex platinum resistance.2. In epithelial ovarian cancer BRCA1 protein in platinum sensitive positive expression is lower than drug-resistant ovarian cancer, Illustrate the BRCA1 protein may be involved in the occurrence of ovarian cancer drug resistance development process.3. HMGB1 protein expression in ovarian tissue, and no difference in platinum sensitive ovarian cancer and ovarian cancer drug resistance, and probably has nothing to do with the process of ovarian cancer drug resistance. HMGB1 expression related to tumor tissue types that HMGB1 may be involved in the pathogenesis of ovarian cancer.4.The expression of P62 was related to clinical stage of ovarian cancer, prompted the P62 may be involved in the development of ovarian cancer; In epithelial ovarian cancer P62 protein in platinum sensitive positive expression is higher than drug-resistant ovarian cancer, Illustrate the P62 protein may be involved in the occurrence of ovarian cancer drug resistance development process.5. There is a negative correlation between the expression of BRCA1 and P62, the BRCA1 positive expression was lower in the higher P62 positive expression. They are related to the progression of ovarian cancer and poor prognosis; BRCA1 and P62 co-expression may predict poor prognosis and instruct postoperative therapy. BRCA1 and P62 can be predicted epithelial ovarian cancer iconic protein of platinum resistance.