应用抗体芯片筛选上皮性卵巢癌患者血清的差异表达蛋白

目的 通过检测健康妇女与上皮性卵巢癌患者血清间蛋白质组表达谱,筛选出差异表达蛋白质,分析异常表达蛋白质的生物学功能.方法 选取11例上皮性卵巢癌患者术前血清混合样品及其配对健康妇女血清混合样品经Cy3和Cy5双色荧光标记后与同一型号的单克隆抗体芯片杂交.根据杂交荧光信号强度,计算内源标准化信噪比(INR),并以INR>2.0或<0.7为临界值,筛选健康妇女与卵巢癌患者术前血清间差异表达的蛋白群.结果 发现27名健康妇女与卵巢癌患者术前血清间明显差异表达的蛋白质,含高表达蛋白16个和低表达蛋白11个.结论 蛋白抗体芯片的研究样品可以是组织,血清等.筛选出的"关键蛋白群",有助于认识卵巢癌发生的分子机理,寻找用于监测卵巢癌进展、判断疗效和预后的分子标志物以及新的治疗靶标.     

《PARP抑制剂在卵巢癌中的应用：ASCO指南》解读

        2014年，美国食品和药物监督管理局（FDA）批准了第一个多聚ADP核糖多聚酶（PARP抑制剂）奥拉帕利（Olaparib）治疗已接受≥3线化疗的胚系BRCA突变（gBRCA）卵巢上皮性癌患者；2016年批准卢卡帕利（Rucaparib）用于治疗胚系和体细胞BRCA突变（g/sBRCA）的复发性卵巢癌患者；2017年批准了尼拉帕利（Niraparib）和Olaparib用于对含铂化疗完全或部分缓解患者的维持治疗。 浏览更多请关注本刊微信公众号及当期杂志。     

Survival impact of capsule rupture in stage I clear cell carcinoma of the ovary in comparison with other histological types

Objective

We analyzed a large number of stage I clear cell carcinoma of the ovary (CCC) patients to estimate the survival impact of the capsule status in stage I CCC patients, particularly in comparison with non-CCC patients.

Methods

Clinicopathologic data on 564 patients with stage I epithelial ovarian cancer (EOC) collected under the central pathological review system were subjected to uni- and multivariable analyses to evaluate the disease-free survival (DFS) and overall survival (OS).

Results

There was no significant difference in both the OS and DFS of CCC patients between IA and IC(ir) (intraoperative capsule rupture) {IA vs. IC(ir); OS: P = 0.1402, DFS: P = 0.2701}. In contrast, CCC patients at IC(non-ir) {IC excluding for IC(ir), such as preoperative capsule rupture, positive ascites/washing, and surface involvement} showed a poorer OS and DFS than those at IC(ir), or those at the corresponding stage in non-CCC. In multivariable analysis, the capsule status was an independent prognostic factor of a poor OS and DFS {OS: HR, 2.832; 95% CI 1.156-6.938; P = 0.023; DFS: HR, 4.327; 95% CI, 1.937-9.667; P = 0.0004)} {In contrast, non-CCC: N.S. (OS/DFS)}. Furthermore, in CCC patients, intraperitoneal recurrences were more frequently observed in IC(non-ir) CCC than IA or IC(ir) CCC (P = 0.0083) {In contrast, non-CCC: N.S.}.

Conclusion

This study suggests that CCC patients other than those with intraoperative capsule rupture show a considerable risk for mortality despite adjuvant chemotherapy.     

血清miRNA-21和miRNA-203在上皮性卵巢癌中的表达及诊断价值

目的 探讨微小核糖核酸21（miR-21）和微小核糖核酸203（miR-203）在上皮性卵巢癌（EOC）中的表达情况,及两者与血清糖类抗原125（CA125）在EOC诊断中的临床价值。方法 采用QPCR检测40例EOC患者（EOC组）、40例上皮性卵巢良性肿瘤患者（良性组）和42例健康女性（正常对照组）血清miR-21和miR-203的表达情况;同时采用电化学发光法检测上述3组人群血清CA125的表达水平;分析miR-21和miR-203的表达与EOC临床病理特征的关系,用受试者工作特征曲线（ROC）评价两者的临床诊断价值并分别计算CA125、miR-21和miR-203单独和联合检测在EOC诊断中的灵敏度和特异度。结果 EOC组miR-21和miR-203的相对表达量分别为2.27±0.48和3.61±0.71,均高于良性组的1.20±0.22和1.47±0.38以及正常对照组的1.00±0.33和1.00±0.28,差异有统计学意义（P＜0.05）;而良性组与正常对照组之间的差异无统计学意义（P＞0.05）。miR-21和miR-203表达与临床分期、淋巴结转移有关（P＜0.05）,与年龄、绝经状态、病理类型和CA125水平无关（P＞0.05）;CA125单独检测的特异度最高,为82.50％;CA125、miR-21和miR-201联合检测的灵敏度最高,达95.00％。结论 miR-21和miR-203可能在EOC的发生、发展中发挥原癌基因的作用;血清CA125、miR-21和miR-203三者联合检测可提高EOC诊断的灵敏度。     

Upregulation of FasL by LPA on ovarian cancer cell surface leads to apoptosis of activated lymphocytes

OBJECTIVE: Constitutive expression and upregulation of FasL by malignant epithelial cells counterattack infiltrating natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) and induce apoptosis of normal cells within the tumor, which may induce metastasis. As little is known about the mechanisms that regulate expression of Fas ligand and the subsequent release of FasL in epithelial ovarian cancer cells (EOC), we investigated the effects of lysophosphatidic acid (LPA) on FasL expression and associated signaling pathways. METHODS: We used established EOC cell lines that were incubated with or without LPA and FasL expression was detected by flow cytometry. Cells were additionally lysed and detected for total protein expression. Activated CD4+ T cells, after coculture with or without EOC, were collected for apoptosis staining and analysis by flow cytometry. RESULTS: Flow cytometry showed that LPA strongly upregulated FasL expression on the OVCAR3 cell surface (P < 0.01), yet in Dov13 cells, LPA significantly upregulated FasL expression only in the presence of the general matrix metalloproteinase (MMP) inhibitors GM6001 and MMP inhibitor II (P < 0.01). The MEK/ERK1/2 kinase cascade is required for FasL upregulation, since the MEK inhibitor PD98059 significantly inhibited FasL upregulation induced by LPA (P < 0.01). Type II secretory phospholipase A2 (sPLA2-II), which promotes protein exocytosis from secretory vesicles and gelatinase granules, affects FasL translocation from intracellular to the cell surface. Pretreatment of Dov13 cells with LPA increased activated T cell apoptosis in cocultures. CONCLUSIONS: These data suggest that upregulation of FasL by LPA provides EOC immune-privilege and leads to apoptosis of activated T lymphocytes.     

Involvement of SDF-1alpha/CXCR4 axis in the enhanced peritoneal metastasis of epithelial ovarian carcinoma

Epithelial ovarian carcinoma (EOC) spreads by implantation of tumor cells onto the human peritoneal mesothelial cells (HPMCs) lining the peritoneal cavity. The aim of this study was to determine whether the stromal cell-derived factor-1alpha (SDF-1alpha)/CXCR4 axis is involved in the interaction of EOC cells with HPMCs in peritoneal metastasis. Clinically, we first evaluated CXCR4 expression in sections from 36 primary EOCs using immunohistochemistry. We next examined whether SDF-1alpha played roles in EOC progression, including in proliferation, cell motility, attachment to HPMCs, and the in vivo development of peritoneal metastasis through CXCR4. Of the 36 carcinomas, 16 cases (44.4%) were positive for CXCR4 immunoexpression. Positive CXCR4 expression significantly predicted poorer overall survival compared with negative expression (p = 0.0069). We found CXCR4 expression in both EOC cells and HPMCs. In contrast, the level of production of SDF-1alpha by HPMCs was higher than that by various EOC cells. Functionally, SDF-1alpha induced enhanced attachment between ES-2 cells and HPMCs or extracellular matrix components. The enhancement of adhesion potential by SDF-1alpha was inhibited by AMD3100, a CXCR4 antagonist, and by phosphatidylinositol 3 kinase and p44/42 inhibitors. Furthermore, intraperitoneal treatment with AMD3100 resulted in reduced dissemination in nude mice inoculated with ES-2 cells. The present results suggest that there may be a link between the SDF-1alpha/CXCR4 axis and enhanced intraperitoneal dissemination of EOC and that CXCR4 may be a novel target for the treatment of EOC.