Tyrosine kinase A receptor (trkA): a potential marker in epithelial ovarian cancer

Objectives

To evaluate the role of trkA receptor as a potential tumor marker in serous epithelial ovarian cancer and its relationship with the angiogenic factors expression as vascular endothelial growth factor (VEGF) and nerve growth factor (NGF). Additionally, to examine whether NGF and VEGF secreted by epithelial ovarian cancer (EOC) explants and from epithelial ovarian cancer cell line (A2780) are involved in the process of angiogenesis, such as cellular proliferation, migration and differentiation of the human endothelial cell line (EA.hy926).

Methods

The mRNA levels of VEGF, NGF and trkA receptors were measured using PCR in 60 ovarian samples. Cellular localization and semi-quantitative estimation of VEGF, NGF, total trkA and p-trkA was performed using IHC in epithelial cells. NGF, total trkA and p-trkA protein were also evaluated in endothelial cells from the same tissues. Human endothelial cell line EA.hy926 was cultured with conditioned media obtained from both EOC explants and from the A2780 cell line, with or without NGF stimulus.

Results

Significantly higher levels of NGF, total trkA and p-trkA protein expressions were observed in epithelial and endothelial cells in poorly differentiated EOC versus normal ovary. Interestingly, the p-trkA receptor expression level showed the most significant difference and its presence was only found in borderline tumor and EOC samples indicating the importance of trkA receptor in EOC as a potential tumor marker.A significant increase in proliferation, migration and differentiation of EA.hy926 cells was observed with NGF, and this effect was significantly reverted when NGF was immuno-blocked and when a trkA inhibitor was used, showing that NGF is an important angiogenic factor in EOC by activating its trkA receptor.

Conclusion

These results indicate that p-trkA may be considered as a new potential tumor marker in EOC, and that NGF may also act as a direct angiogenic factor in EOC.     

sTWEAK在上皮性卵巢癌患者血清中的表达及其意义

目的:检测卵巢上皮性癌(EOC)患者外周血可溶性肿瘤坏死因子样凋亡微弱诱导剂(sTWEAK)的表达水平,并分析其临床意义。方法:选取2010年1月至2014年2月在上海交通大学附属仁济医院住院行手术的上皮性卵巢癌(EOC)患者和良性上皮性卵巢肿瘤患者共43例,采用ELISA法检测外周血血清中sTWEAK的表达水平。结果:(1)与良性上皮性卵巢肿瘤组相比,EOC组的sTWEAK的表达水平明显升高(P0.05)。两组患者中sTWEAK的表达与CA125呈正相关(r=0.42,P0.05),其中CA125500U/ml者的sTWEAK表达水平明显低于CA125500U/ml者(P0.05)。(2)EOC患者血清中,sTWEAK的表达水平与其临床分期、病理分级、组织类型以及腹水有无肿瘤细胞均无相关性(P0.05)。(3)EOC组中,CA125500U/ml者行满意肿瘤细胞减灭术的比例显著高于CA125500U/ml者(P0.05)。结论:sTWEAK虽不能反映EOC患者的肿瘤进展,但其与EOC的发生密切相关,sTWEAK可能成为判断卵巢肿瘤良恶性的新型血清学标记,在EOC诊断及预后预测中具有一定的应用前景。     

Gene expression in epithelial ovarian cancer: a study of intratumor heterogeneity

The aim of this study was to investigate the intratumor heterogeneity of gene expression profiles in epithelial ovarian cancer (EOC). This was done to evaluate whether sampling of a single macrodissected tissue sample from each EOC case would bias the data and result in, eg, prognostic studies based on gene expression microarray experiments. From nine EOCs removed at Odense University Hospital, Denmark, three tumor samples of 200-300 mg each were taken with greatest possible mutual distance. The samples were immediately flash frozen. A parallel section was taken for histopathologic comparison. RNA was extracted from the tissue samples. Five micrograms of each RNA sample was used for labeling. The fragmented biotin-labeled complementary RNA was hybridized to Affymetrix GeneChip Human Genome U133 plus 2.0 arrays, and scanning was performed on the GeneArray scanner 3000 (Affymetrix, Santa Clara, CA). Data were evaluated using hierarchical clustering and intraclass correlation coefficient (ICC) from reliability analysis. All evaluation methods revealed low intratumor heterogeneity. Intratumor ICCs ranged from 0.888 to 0.978. In contrast, "between-tumor" ICC was 0.549 indicating much lower intra- than intertumor heterogeneity. Due to a low degree of intratumor variation, we conclude that it is sufficiently accurate in a clinical setup to use single, macrodissected tumor samples in the evaluation of gene expression in EOCs.     

Spontaneous regression in recurrent epithelial ovarian cancer

Background Although spontaneous regression has been reported in several cancers, it is generally believed that this rare phenomenon does not occur in epithelial ovarian cancer. Case We presented a stage IV epithelial ovarian cancer patient with long-term disease-free survival after palliative local irradiation alone against recurrence. The recurrent supraclavicular lymph node was almost completely necrotic, and the swelling of the para-aortic lymph node spontaneously regressed. Conclusion The histologic features and unexpected clinical course in this patient strongly suggest that spontaneous regression occurred in recurrent epithelial ovarian cancer.     

Frequency of serous tubal intraepithelial carcinoma (STIC) in patients with high grade serous ovarian cancer

ObjectiveSerous tubal intraepithelial carcinoma (STIC) is a known precursor of high-grade serous ovarian cancer (HGSOC). This study aimed to evaluate the proportion of STIC in patients with HGSOC and analyze the STIC-related prognosis in patients with HGSOC.Materials and methodsAll pathology reports at our institution that included bilateral salpingectomies of patients with HGSOC from January 2013 to December 2018 were reviewed by two experienced pathologists. The specimens from the ovaries and the salpinx including fimbria were examined. We analyzed the correlation between STIC and HGSOC and compared the clinical characteristics and STIC-related prognostic outcomes in patients with HGSOC.ResultsEleven of the 76 cases were STIC. BRCA mutations were found in 16.9% of patients with HGSOC. STIC was observed in 30.0% of patients with BRCA mutations and in 14.3% of patients without BRCA mutations. The incidence of STIC in patients with BRCA mutations was approximately twice that in patients without BRCA mutations; however, the difference was not statistically significant (P = 0.231). Further, the 5-year survival rate of patients without STIC appeared to be high; nevertheless, the difference was not statistically significant (59.7% vs. 47.4%, P = 0.633). Moreover, there was no significant difference in disease-free survival rate according to STIC (36.4% vs. 33.1%, P = 0.956).ConclusionSTIC was identified in patients with HGSOC, and STIC incidence was prominent in HGSOC related to BRCA mutation. Although low frequency, STIC was detected in patients without BRCA mutation. Therefore, prophylactic salpingectomy may be useful for prevention of HGSOC.     

The search for biomarkers to direct antiangiogenic treatment in epithelial ovarian cancer

Antiangiogenic agents have demonstrated improved progression-free survival in women with primary and recurrent epithelial ovarian cancer (EOC). Biomarkers that predict outcomes in patients treated with antiangiogenic agents are being investigated to rationally direct therapy for women most likely to benefit from these agents. Among the most promising plasma-based biomarkers are vascular endothelial growth factor (VEGF)-A, fibroblast growth factor, platelet-derived growth factor, angiopoietin-2, and VEGF receptor-2. While these biomarkers have been correlated with prognosis, they have not been shown to predict benefit, specifically from anti-VEGF therapy, highlighting the need for alternative biomarkers, including molecular and clinical factors, which may be predictive of outcome in women with ovarian cancer treated with antiangiogenic agents. Biomarkers are currently being investigated as secondary outcomes in several ongoing phase II and phase III clinical trials of antiangiogenic agents in patients with EOC. Molecular techniques, such as microarray analyses, and imaging techniques, such as dynamic contrast-enhanced magnetic resonance imaging, positron emission tomography, and single photon emission computed tomography, are also being explored in this field. In this review, we provide a comprehensive overview of current biomarker research, with an emphasis on angiogenic biomarkers associated with EOC.     

Defective expression of Protein 4.1N is correlated to tumor progression,aggressive behaviors and chemotherapy resistance in epithelial ovarian cancer

Objective

Protein 4.1N (4.1N) is a member of the Protein 4.1 family that is involved in cellular processes such as cell adhesion, migration and signaling. In this study, we evaluated the expression of 4.1N protein and its potential roles in epithelial ovarian cancer (EOC) tumorigenesis and progression.

Methods

4.1N protein expression was investigated in a total of 280 samples including 74 normal tissues, 35 benign, 30 borderline and 141 malignant epithelial ovarian tumors by immunohistochemistry. Correlation between 4.1N expression levels and clinicopathologic features was statistically analyzed. The expression of 4.1N in EOC cell lines was examined by western blotting.

Results

Immunohistochemistry analysis revealed that, although there was no loss of 4.1N expression in normal tissues and benign tumors, absence of Protein 4.1N was significantly more common in EOCs (44.0%) than in borderline tumors (3.3%) (p < 0.001). Furthermore, loss or decreased expression of 4.1N protein expression was correlated with malignant potential of the tumors (14.3% in benign tumors, 56.7% in borderline tumors and 92.9% in malignancy) (p < 0.001). In EOC samples, loss of 4.1N protein was significantly associated with advanced-stage (p = 0.004), ascites (p = 0.009), omental metastasis (p = 0.018), suboptimal debulking (p = 0.024), poorly histological differentiation (p = 0.009), high-grade serous carcinoma (p = 0.001), short progression-free-survival (p = 0.018) and poor chemosensitivity to first-line chemotherapy (p = 0.029). Moreover, western blotting analysis revealed that expression of 4.1N protein was lost in 4/8 (50%) EOC cell lines.

Conclusions

4.1N protein expression level was significantly decreased during malignant transformation of epithelial ovarian tumors and that loss of 4.1N expression was closely correlated to poorly differentiated and biologically aggressive EOCs.     

微小RNA-7在上皮性卵巢癌原发灶及转移灶中表达的研究

目的:检测上皮性卵巢癌组织原发灶、转移灶中miR-7及其靶标EGFR蛋白的表达,探讨miR-7及EGFR与卵巢癌转移的关系。方法:采用显色原位杂交(CISH)检测miR-7在卵巢癌组织原发灶、转移灶石蜡切片中的表达;采用免疫组化检测EGFR蛋白表达。结果:上皮性卵巢癌转移灶中miR-7表达显著低于原发灶(P0.05),而EGFR表达显著高于原发灶(P0.05);miR-7与EGFR表达具有相关性(r=-0.441,P0.05)。结论:miR-7可能通过EGFR抑制卵巢癌的转移,miR-7可能成为治疗卵巢癌的新靶标。     

Prognostic significance of human epididymis protein 4 in epithelial ovarian cancer

Objective

The purpose of this study was to evaluate the prognostic significance of serum human epididymis protein 4 (HE4) level in patients with epithelial ovarian cancer.

Study design

A total of 78 women diagnosed with a pelvic mass and operated on in our institute comprised our cohort. Forty-five of these were diagnosed with epithelial ovarian cancer and treated with debulking surgery, followed by taxane and platinum-based chemotherapy as clinically indicated. Preoperatively obtained serum samples were analyzed for levels of HE4 and CA125.

Results

The elevated serum HE4 level was related to advanced stage and serous type of cancer. The median duration of the follow-up was 35.1 months. In advanced stage, the median progression-free survival (PFS) of patients with elevated serum HE4 levels was 20.1 months (95% CI, 15.7–24.6 months), whereas that of patients with normal serum HE4 level was 24.2 months (95% CI, 13.9–34.6 months) (p = 0.029). Independent predictors for PFS in patients with advanced stage EOC included serum HE4 level (hazard ratio 2.24; 95% CI, 1.14 to 6.84; p = 0.048).

Conclusions

Our results demonstrated that an elevated serum HE4 level was related to the advanced stage of epithelial ovarian cancer. An elevated serum level of HE4 is a poor prognostic factor for PFS in patients with epithelial ovarian cancer who were treated with debulking surgery and adjuvant taxane and platinum-based chemotherapy. The serum HE4 level is a promising indicator for the progression of cancer as well as a biomarker for the detection of epithelial ovarian cancer.     

The expression levels and clinical significance of MFG-E8 and CD133 in epithelial ovarian cancer

Abstract

The aim of our study was to test whether there is an association between high expression of milk fat globule EGF factor 8 (MFG-E8) and CD133 presence or clinical outcomes of patients with epithelial ovarian cancer (EOC). MFG-E8 and CD133 expression levels were analyzed by immunohistochemistry in 88 EOC tumor specimens. High expression of MFG-E8 directly and significantly correlated with the presence of CD133 immunostaining (R?=?0.353, p=.001), whereas immunostaining of MFG-E8 and CD133 significantly correlated with FIGO stage, tumor grade, debulking status, the dualistic model, ascites status, and nonresponse to chemotherapy (p<.05). It was also found that high expression of MFG-E8 and CD133 presence is a potent predictor of poor clinical outcomes among patients with EOC. Our study is the first to show that high expression of MFG-E8 in EOCs positively correlates with CD133 presence. Further research on MFG-E8 in EOC is needed to determine whether MFG-E8 is a new tumor marker of ovarian cancer and a new target for anticancer therapy as well as whether it can interact with cancer stem cell inhibitors for the treatment of refractory tumors.     

Overexpression of c-Abl predicts unfavorable outcome in epithelial ovarian cancer

Objectives

Abelson tyrosine kinase (c-Abl) has been shown to promote solid tumor invasion and metastasis. However, little is known regarding whether c-Abl contributes to the development or progression of epithelial ovarian cancer (EOC). The aims of this study are to determine the expression of c-Abl and investigate a possible relationship between c-Abl and prognosis in EOC.

Methods

c-Abl protein level was evaluated in 137 EOC specimens by immunohistochemical staining and 32 EOC specimens by Western blot analysis. Expression of c-Abl in ovarian cancer cell lines was measured by Western blot analysis and immunofluorescence. Survival analysis was performed to assess the correlation between c-Abl expression and survival.

Results

Immunohistochemical staining and Western blot analysis revealed that c-Abl was overexpressed in EOC compared with samples from a non-invasive ovarian tumor and normal ovaries (P < 0.05). Furthermore, expression of c-Abl was significantly associated with advanced FIGO stage, poor grade, serum Ca-125 and residual tumor size (P < 0.05). By Western blot analysis, c-Abl expression was examined in four ovarian cancer cell lines. Meanwhile, immunofluorescence was performed to show c-Abl expression in SKOV3 and 3AO cell lines. Survival analysis demonstrated that patients with low c-Abl staining had a significantly better survival compared to patients with high c-Abl staining (P < 0.05). In multivariate analysis, c-Abl overexpression, poor grade, advanced stage and suboptimal surgical debulking were independent prognostic factors of poor survival.

Conclusions

Our present study finds that c-Abl overexpression is associated with an unfavorable outcome. c-Abl may be a crucial predictor for EOC metastasis.     

Alcohol drinking and epithelial ovarian cancer risk. a systematic review and meta-analysis

Objective

In order to provide an updated quantification of the association between alcohol drinking and epithelial ovarian cancer risk, we conducted a meta-analysis of published observational studies.

Methods

Using PubMed, we performed a literature search of all case-control and cohort studies published as original articles in English up to September 2011. We included 27 observational studies, of which 23 were case-control studies, 3 cohort studies and one pooled analysis of prospective cohort studies, including a total of 16,554 epithelial ovarian cancer cases. We derived pooled meta-analytic estimates using random-effects models.

Results

The pooled relative risk (RR) for any alcohol drinking compared with non/occasional drinking was 1.00 [95% confidence interval (CI), 0.95-1.05]. The RRs were 0.97 (95% CI, 0.92-1.02), 1.03 (95% CI, 0.96-1.11) and 1.09 (95% CI, 0.80-1.50) for light (≤ 1 drink/day), moderate (> 1 to < 3 drinks) and heavy drinking (≥ 3 drinks/day), respectively. In particular, the pooled RR for invasive epithelial ovarian cancers was 1.00 (95% CI, 0.95-1.06), while for borderline cancers was 0.96 (95% CI, 0.74-1.26). Stratified analyses across cancer histotypes revealed a modest protective effect of alcohol on endometrioid epithelial ovarian tumors (RR = 0.82, 95% CI, 0.70-0.96), while no association was found for serous (RR = 1.00, 95% CI, 0.84-1.19), mucinous (RR = 0.91, 95% CI, 0.78-1.08) and clear cell (RR = 0.93, 95% CI, 0.76-1.14) cancers. There was no evidence of publication bias.

Conclusions

This comprehensive meta-analysis provided no evidence of a material association between alcohol drinking and epithelial ovarian cancer risk.     

MAGE-A family serves as poor prognostic markers and potential therapeutic targets for epithelial ovarian cancer patients: a retrospective clinical study

Objective: The aim of our study is to investigate the expression pattern and prognostic significance of melanoma-associated antigens-A (MAGE-A) family in primary epithelial ovarian cancer (EOC) patients.

Study design: The expression of MAGE-A family members, including MAGE-A1, -A2, -A3, -A4, -A6, -A10 and -A12 was immunohistochemically detected in 82 cases of primary EOC and 10 cases of pericarcinoma ovarian tissues. The association between MAGE-A family expression and the clinicopathological parameters as well as the prognosis of primary EOC patients was analyzed.

Results: MAGE-A family expressed in 48.8% of primary EOC tissues, but not expressed in pericarcinoma ovarian tissues. MAGE-A expression was associated with the pathological types, FIGO stage, and pre-operative serum CA125 level. Overall survival of EOC patients with positive MAGE-A family expression was significantly shorter than those patients with negative MAGE-A expression. Multivariate analysis showed that although MAGE-A family expression can affect the overall survival, it was not an independent prognostic marker for EOC patients.

Conclusions: Molecular assessment of MAGE-A family members could be helpful to improve the prognostic evaluation and to provide a new potential therapeutic target for primary EOC patients.     

雄激素受体基因CAG多态性与卵巢早衰的关系

目的:探讨雄激素受体(AR)基因CAG重复序列与特发性卵巢早衰(POF)发病的关系.方法:选择特发性POF患者85例为研究组,健康女性80例为对照组.提取两组患者的外周血DNA,PCR扩增并采用琼脂糖凝胶电泳获取目的基因,进行基因测序,检测每例AR基因CAG重复序列的重复次数.结果:POF组与对照组的最大频率等位基因分别为n=23及n=22;(CAG)n重复次数分别为22.73±3.24、22.03±3.06,差异具有显著性(P＜0.05);POF组(CAG)n长片段(n≥23)的发生频率显著高于对照组(P＜0.05).结论:POF患者AR基因CAG重复次数增多,CAG长片段的发生频率增高.AR基因CAG重复序列次数多态性可能与中国汉族女性的POF发病有关.     

卵巢浆液性癌组织中Hugl-1和aPKC的表达及意义

目的探讨原发性卵巢浆液性癌组织中Hugl-1和aPKC的表达及意义。方法选择浙江大学医学院附属妇产科医院2000年6月至2007年6月经手术切除的原发性卵巢浆液性癌组织79例,用免疫组化SP法检测Hugl-1和aPKC蛋白在其组织中的表达。同时检测Hugl-1和aPKC蛋白在25例正常卵巢组织,31例卵巢浆液性囊腺瘤,41例卵巢交界性浆液性肿瘤中的表达,并分析了Hugl-1和aPKC蛋白表达与卵巢癌临床病理参数间的关系。结果Hugl-1蛋白在卵巢浆液性癌组织、正常卵巢、浆液性囊腺瘤及交界性浆液性肿瘤中的强阳性表达率分别为12.7%、48%、58%、41.5%。癌组织中的表达显著低于其余3组,差异有统计学意义(P0.001);aPKC蛋白在卵巢浆液性癌组织、正常卵巢、浆液性囊腺瘤及交界性浆液性肿瘤中的强阳性表达率分别为19%、0、0、0。癌组织中的表达显著高于其余3组,差异有统计学意义(P0.001);Hugl-1和aPKC的表达与组织学分级、手术病理分期、淋巴结转移有一定的相关性(P0.05)。Hugl-1和aPKC在卵巢癌组织中的表达不存在相关性(P0.05)。结论Hugl-1和aPKC可能均参与了卵巢癌的发生与发展。     

上皮性卵巢癌PTEN蛋白表达分析

目的：探讨抑癌基因PTEN的蛋白表达与上皮性卵巢癌的发生、发展之间的关系。方法：对74例上皮性卵巢癌应用免疫组化方法检测PTEN蛋白的表达。结果：上皮性卵巢癌中PTEN蛋白有一定的表达缺失，在G1、G2级组织表达率（90．62％）高于G3级组织表达率（66．67％），二者差异有显著性；不同的临床分期中，I、Ⅱ期阳性表达为22／24（91．67％），Ⅲ、Ⅳ期阳性表达为35／50（70％），二者差异有显著性。在浆液性和粘液性两种不同的组织类型间，表达率分别为82．98％和66．67％，二者无明显差别。结论：PTEN蛋白的表达缺失在上皮性卵巢癌的发生、发展过程中有一定的作用，且与病变进展和分化有关。