PAX8、PAX2、p53和RAS蛋白在卵巢及输卵管组织中的表达对于高级别卵巢浆液性癌起源的意义

Significance and expression of PAX8, PAX2, p53 and RAS in ovary and fallopian tubes to origin of ovarian high grade serous carcinoma

目的探讨配对盒基因8抗体（PAX8）、配对盒基因2抗体（PAX2）、p53和ras癌基因（RAS）蛋白在高级别卵巢浆液性癌（HGSC）患者的卵巢及输卵管组织中的表达，并分析其对于HGSC起源的意义。 方法 （1）组织标本：收集2015年1月—2016年1月广西医科大学附属肿瘤医院收治的34例卵巢恶性肿瘤患者的组织标本，以同期因卵巢良性疾病等行手术治疗患者的卵巢和输卵管无病变区域的组织标本10份作为对照。其中，卵巢组织标本的取材按左、右侧区分，输卵管组织标本的取材按左、右侧区分并按伞部、壶腹部、峡部解剖分段取材。34例卵巢恶性肿瘤患者中，上皮性恶性肿瘤29例（包括浆液性癌27例、黏液性癌1例、子宫内膜样腺癌1例）、非上皮性恶性肿瘤5例（均为性索-间质肿瘤）；27例卵巢浆液性癌患者中，HGSC 23例、低级别卵巢浆液性癌（LGSC）4例。（2）组织芯片：采用由山东大学齐鲁医院制作并赠送的组织芯片，该芯片收集了2005—2013年手术切除并经病理诊断明确为卵巢浆液性癌的组织标本153份。采用免疫组化SP法检测上述组织标本和组织芯片中PAX8、PAX2、p53和RAS蛋白的表达，（1）分析HGSC患者和对照妇女的卵巢和输卵管组织中PAX8、PAX2、p53及RAS蛋白的定位表达差异；（2）比较不同病理类型卵巢恶性肿瘤患者的卵巢和输卵管组织中PAX8、PAX2、p53及RAS蛋白表达水平的差异；（3）分析PAX8、PAX2、p53、RAS蛋白在HGSC患者输卵管组织中的表达水平与其对应卵巢组织中表达水平的相关性；（4）采用单因素生存分析法对组织芯片中卵巢浆液性癌患者的预后影响因素进行分析。结果（1）对照妇女的正常卵巢上皮细胞中PAX8、PAX2、p53、RAS蛋白均呈阴性表达，而正常输卵管上皮分泌型细胞中PAX8、PAX2、p53、RAS蛋白均呈棕褐色强阳性表达。HGSC患者的输卵管上皮分泌型细胞中及卵巢和输卵管的癌细胞中PAX8、PAX2、p53、RAS蛋白均呈棕褐色强阳性表达。（2）上皮性卵巢恶性肿瘤患者的输卵管组织中p53、RAS蛋白的表达水平均明显高于非上皮性卵巢恶性肿瘤患者（P〈0.05）；但两者的输卵管组织中PAX8、PAX2蛋白以及卵巢组织中PAX8、PAX2、p53、RAS蛋白的表达水平比较均无明显差异（P〉0.05）。HGSC患者的卵巢组织中PAX8、PAX2和p53蛋白的表达水平均明显高于LGSC患者（P〈0.05），RAS蛋白的表达水平明显低于LGSC患者（P〈0.05）；而两者的输卵管组织中PAX8、PAX2、p53、RAS蛋白的表达水平分别比较均无明显差异（P〉0.05）。（3）PAX8、PAX2蛋白在HGSC患者输卵管组织中的表达水平与其对应卵巢组织中的表达水平均呈明显正相关（r=0.422，P=0.045；r=0.693，P=0.000）；但p53和RAS蛋白在输卵管组织中的表达水平与其对应卵巢组织中的表达水平均无相关性（r=0.058，P=0.793；r=-0.190，P=0.384）。（4）单因素生存分析显示，卵巢浆液性癌患者的无进展生存时间与PAX8、PAX2、RAS蛋白表达均明显相关（P〈0.05）；而与年龄、手术病理分期、病理分化程度、淋巴结转移状态、术前化疗以及p53蛋白表达均无关（P〉0.05）。卵巢浆液性癌患者的总生存时间与PAX8蛋白表达有关，而与年龄、手术病理分期、病理分化程度、淋巴结转移状态、术前化疗以及PAX2、p53、RAS蛋白表达均无关（P〉0.05）。结论PAX8、PAX2、p53、RAS蛋白对于研究HGSC的起源定位于输卵管上皮分泌型细胞有着重要的意义，HGSC可能来源于输卵管，但需更深入的研究来证实；PAX8、PAX2、p53、RAS蛋白有望作为卵巢浆液性癌患者预后的预测指标。

To explore the origin of ovarian high grade serous carcinoma （HGSC） through analysing the expression and significance of PAX8, PAX2, p53 and RAS in the ovary and fallopian tube of different types and grades of serous carcinoma.Methods A total of 44 cases tissue samples of ovarian tumor including 34 malignant ovarian tumor and 10 normal normal tissue （as control group） were collected from the admitted patients in Affiliated Tumor Hospital of Guangxi Medical University from January 2015 to January 2016. Fallopian tube tissues were segmented in accordance with the fimbria, ampulla, isthmus and the corresponding ovarian tissues were by the side. There were 34 cases of patients with ovarian cancer including 29 cases of epithelial ovarian cancer （27 serous carcinoma, 1 mucinous carcinoma,1 endometrioid adenocarcinoma） and 5 non-epithelial ovarian cancer （sex cord-interstitial tumor）. Among 27 cases of patients with ovarian serous cancer, there were 23 HGSC and 4 low-grade ovarian serous cancer （LGSC）. One hundred fifty-three cases of samples were diagnosed as ovarian serous cancer by Shandong University Affiliated Qilu Hospital from 2005 to 2013 and these samples were made tissue microarray. （1） To analyze the expression and differences of PAX8, PAX2, p53 and RAS in the above tissues and tissue microarray from ovarian and tubal of HGSC and control women by immunohistochemistry methods. （2） To compare the expression levels of PAX8, PAX2, p53 and RAS in ovarian and fallopian tubes of ovarian cancer patients with different pathological types. （3） To analyze the correlations of tubal and ovarian tissue in PAX8, PAX2, p53 and RAS expression of HGSC. （4） To analyze the factors of the prognosis of ovarian serous cancer in tissue microarray by single factor analysis method.Results （1） PAX8, PAX2, p53 and RAS expression was negative in normal ovarian epithelium of control group, but the expression of PAX8, PAX2, p53 and RAS were strongly positive brown in secrete cells of normal fallopian tube epithelium. （2） p53 and RAS expression of fallopian tube epithelium in the epithelial ovarian cancer group were significantly higher than those in the non-epithelial ovarian cancer groups （P〈0.05）, but the expression of PAX8 and PAX2 in fallopian tube and the expression of PAX8, PAX2, p53 and RAS in ovarian tissue was not statistically significant in the groups （P〉0.05）. PAX8, PAX2 and p53 expression of the ovarian in HGSC group were significantly higher than those in LGSC group （P〈0.05）, while the expression of RAS was lower in the ovarian of the high-grade group （P〈0.05）, while the expression of PAX8, PAX2, p53 and RAS in fallopian tube was not statistically significant in the groups （P〉0.05）. （3） There was a significantly positive correlation between fallopian tube and the corresponding ovary of HGSC in PAX8 and PAX2 expression （r=0.422, P=0.045; r=0.693, P=0.000）, but not correlation in p53 and RAS expression （r=0.058, P=0.793; r= -0.190, P=0.384）. （4） Univariate survival analysis showed that the progression free survival time in patients with ovarian serous cancer group was significantly correlated with the protein expression of PAX8, PAX2 and RAS （P〈0.05）, but there were not correlated with age, surgical staging, cell differentiation, lymph node metastasis and preoperative chemotherapy and p53 protein expression （P〉0.05）. The total survival time in patients with ovarian serous cancer group was significantly correlated with the protein expression of PAX8 （P〈0.05）, but there were not correlated with age,surgical staging, cell differentiation, lymph node metastasis and preoperative chemotherapy and the protein expression of PAX2, RAS and p53 （P〉0.05）. Conclusions PAX8, PAX2, p53, RAS are of great significance for the study of origin of HGSC. HGSC may be derived from fallopian tube, but further investigation would be necessary to confirm this. PAX8, PAX2, p53, RAS could be expected to be used as predictors of survival prognosis in patients with ovarian serous cancer.