Genomic profiling of the residual disease of advanced high-grade serous ovarian cancer after neoadjuvant chemotherapy

The goal of our study was to demonstrate the spectrum of genomic alterations present in the residual disease of patients with advanced high-grade serous ovarian cancer (HGSOC) after neoadjuvant chemotherapy (NAC), including matched pretreatment biopsies. During the study period between 2006 and 2017, we collected pre-NAC and post-NAC tumor tissue samples from patients with advanced HGSOC. We performed combined next-generation sequencing and immunohistochemistry to identify actionable targets and pathway activation in post-NAC residual tumors. We also examined whether post-NAC profiling of residual HGSOC identified targetable molecular lesions in the chemotherapy-resistant component of tumors. Among 102 post-NAC samples, 41 (40%) of patients had mutations in homologous recombination repair (HRR) genes (HRR deficiency). Patients with HRR mutations had higher tumor mutation burdens (p < 0.001) and higher alterations in the PI3K–AKT–mTOR pathway (p = 0.004) than patients without these HRR mutations. Nevertheless, we found no significant differences in progression-free survival (p = 0.662) and overall survival (OS; p = 0.828) between the two groups. Most patients (91%) had alterations in at least one of the targetable pathways, and those patients with cell cycle (p = 0.004) and PI3K–AKT–mTOR signaling (p = 0.005) pathway alterations had poorer OS (Bonferroni-corrected threshold = 0.0083, 0.05/6). We showed the genomic landscape of tumor cells remaining in advanced HGSOC after NAC. Once validated, these data can help inform biomarker-driven adjuvant studies in targeting residual tumors to improve the outcomes of patients with advanced HGSOC after NAC.     

miR -542-5p 在浆液性卵巢癌中的表达及意义

目的：检测 miR -542-5p 在浆液性卵巢癌中的表达,并分析 miR -542-5p 的表达与浆液性卵巢癌临床病理特征之间的关系,探讨其意义。方法：采用实时荧光定量 PCR 技术分析100例浆液性卵巢癌组织及50例正常输卵管伞端组织中 miR -542-5p 的表达。结果：浆液性卵巢癌组织中 miR -542-5p 的表达显著低于正常输卵管伞端组织（P ﹤0.05）。miR -542-5p 的表达与浆液性卵巢癌各临床病理特征之间差异均无统计学意义（P ﹥0.05）。结论：miR -542-5p 可能作为抑癌基因参与了浆液性卵巢癌的发生,对于卵巢癌的早期诊断及治疗具有潜在意义。     

Similar protein expression profiles of ovarian and endometrial high-grade serous carcinomas

Background:

Ovarian and endometrial high-grade serous carcinomas (HGSCs) have similar clinical and pathological characteristics; however, exhaustive protein expression profiling of these cancers has yet to be reported.

Methods:

We performed protein expression profiling on 14 cases of HGSCs (7 ovarian and 7 endometrial) and 18 endometrioid carcinomas (9 ovarian and 9 endometrial) using iTRAQ-based exhaustive and quantitative protein analysis.

Results:

We identified 828 tumour-expressed proteins and evaluated the statistical similarity of protein expression profiles between ovarian and endometrial HGSCs using unsupervised hierarchical cluster analysis (P<0.01). Using 45 statistically highly expressed proteins in HGSCs, protein ontology analysis detected two enriched terms and proteins composing each term: IMP2 and MCM2. Immunohistochemical analyses confirmed the higher expression of IMP2 and MCM2 in ovarian and endometrial HGSCs as well as in tubal and peritoneal HGSCs than in endometrioid carcinomas (P<0.01). The knockdown of either IMP2 or MCM2 by siRNA interference significantly decreased the proliferation rate of ovarian HGSC cell line (P<0.01).

Conclusions:

We demonstrated the statistical similarity of the protein expression profiles of ovarian and endometrial HGSC beyond the organs. We suggest that increased IMP2 and MCM2 expression may underlie some of the rapid HGSC growth observed clinically.     

Clinicodemographic factors influencing outcomes in patients with low-grade serous ovarian carcinoma

BACKGROUND:

Low‐grade serous carcinoma (LGSC) of the ovary is a rare tumor that is distinct from its high‐grade counterpart. The objective of this study was to determine whether patient demographic factors and clinical treatment histories affected survival in a population of women with LGSC.

METHODS:

A review of patients who had pathologically confirmed LGSC of the ovary diagnosed between 1977 and 2009 was performed. Abstracted data included medical and social histories, anthropometric measurements, and details about diagnosis, treatment, and follow‐up. Statistical analyses included Fisher exact tests, Cox proportional hazards models, and the Kaplan‐Meier method.

RESULTS:

The study sample included 194 patients who had a median follow‐up of 60.9 months (range, 1‐383 months). In multivariate analyses, smoking had a negative association with both overall survival (OS) (hazard ratio [HR], 1.73; 95% confidence interval [CI], 1.03‐2.92; P = .04) and progression‐free survival (PFS) (HR, 1.72; 95% CI, 1.00‐2.96; P = .05). The median OS was shorter in current smokers than in former/never smokers (48.0 months vs 79.9 months; P = .002). PFS also was predicted by year of diagnosis >1994 (HR, 1.74; P = .01). Although the difference was not statistically significant, hormone consolidation appeared to be associated with better OS (HR, 0.15; P = .06) and better PFS (HR, 0.44; P = .07). A smaller proportion of the patients who received hormone consolidation experienced disease recurrence compared with the patients who did not receive hormone consolidation (66.7% vs 87.6%; P = .07).

CONCLUSIONS:

Smoking was associated negatively with survival outcomes in women with LGSC of the ovary, whereas consolidation treatment with hormone antagonists demonstrated a protective associative trend with survival. Both lifestyle modification and innovative treatment plans should be considered in this group of patients. Cancer 2011. © 2011 American Cancer Society.     

High-temperature-required protein A2 as a predictive marker for response to chemotherapy and prognosis in patients with high-grade serous ovarian cancers

Background:

High-temperature-required protein A2 (HtrA2), a protein relating with apoptosis in a caspases-dependent and non-dependent manner, has been reported to be associated with chemosensitivity in several human cancers.

Methods:

Tissue microarrays made from 142 patients with high-grade serous ovarian adenocarcinoma were evaluated to assess whether HtrA2 expression was related with several clinical parameters.

Results:

Negative HtrA2 expression was observed in 36 cases (25%) of the patients, and related with significantly lower response rates of primary chemotherapy than those with positive HtrA2 expression (56% vs 83%, P<0.01). In addition, negative HtrA2 expression was identified as an independent worse prognostic factor for progression-free survival and overall survival by multivariate analyses. Furthermore, HtrA2 downregulation modulated sensitivity to platinum in serous ovarian cancer cells in vitro.

Conclusions:

HtrA2 expression was a predictor for sensitivity to chemotherapy, and could be a candidate of molecular target in the treatment of high-grade serous ovarian cancers.     

Key nodes of a microRNA network associated with the integrated mesenchymal subtype of high-grade serous ovarian cancer

Metastasis is the main cause of cancer mortality. One of the initiating events of cancer metastasis of epithelial tumors is epithelial-to-mesenchymal transition (EMT), during which cells dedifferentiat...     

肝细胞癌组织microRNA-187-3p表达预后意义

目的 微小RNA (microRNA,miRNA)差异表达与多种肿瘤的发生发展密切相关,但微小RNA-187-3p(miR-187-3p)在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及预后意义研究较少.本研究检测HCC组织中microRNA的表达谱,分析miR-187-3p在HCC组织中的表达及其与临床病理特征之间的关系,探讨miR-187-3p表达在其发生、发展及预后中的作用.方法 利用microRNA芯片杂交技术筛选5例HCC组织及相应癌旁组织中差异表达的microRNA,然后通过实时荧光定量PCR(real-time fuorescence quantitative-PCR,RFQ-PCR)法验证86例HCC组织及相应癌旁组织中miR-187-3p表达水平,并统计临床病理资料,随访生存期.结果 148个microRNAs在HCC组织中差异表达,其中102个microRNAs在HCC组织较相应癌旁组织表达下调,46个microRNAs在HCC组织较相应癌旁组织表达上调.RFQ-PCR结果显示,miR-187-3p在HCC组织中表达水平明显低于相应癌旁组织,差异有统计学意义,Z=-2.244,P=0.025.HCC中miR-187-3p表达状态与肿瘤大小(x2=5.031,P=0.025)相关,而与性别(x2=3.648,P=0.056)、年龄(x2=0.003,P=0.956)、是否转移(x2=0.005,P=0.943)、TNM分期(x2=0.129,P=0.719)、肿瘤数目(x2 =0.126,P=0.722)、乙肝表面抗原(x2=0.019,P=0.890)、AFP水平(x2=0.187,P=0.665)和Child分级(x2=1.665,P=0.197)无关.miR-187-3p低表达的HCC患者总生存期较高表达者明显缩短,x2=7.684,P=0.006.且多因素分析发现,miR-187-3p表达水平和TNM分期是影响HCC患者生存预后的独立危险因素.结论 miR-187-3p在一定程度上参与了HCC的发生发展,其有望成为HCC新的潜在的治疗靶点以及诊断、判断病情和预测预后的分子标志物.     

Multisite analysis of high-grade serous epithelial ovarian cancers identifies genomic regions of focal and recurrent copy number alteration in 3q26.2 and 8q24.3

High-grade serous epithelial ovarian cancer (HGS-EOC) is a systemic disease, with marked intra and interpatient tumor heterogeneity. The issue of spatial and temporal heterogeneity has long been overlooked, hampering the possibility to identify those genomic alterations that persist, before and after therapy, in the genome of all tumor cells across the different anatomical districts. This knowledge is the first step to clarify those molecular determinants that characterize the tumor biology of HGS-EOC and their route toward malignancy. In our study, -omics data were generated from 79 snap frozen matched tumor biopsies, withdrawn before and after chemotherapy from 24 HGS-EOC patients, gathered together from independent cohorts. The landscape of somatic copy number alterations depicts a more homogenous and stable genomic portrait than the single nucleotide variant profile. Genomic identification of significant targets in cancer analysis identified two focal and minimal common regions (FMCRs) of amplification in the cytoband 3q26.2 (region α, 193 kb long) and 8q24.3 (region β, 495 kb long). Analysis in two external databases confirmed regions α and β are features of HGS-EOC. The MECOM gene is located in region α, and 15 genes are in region β. No functional data are yet available for the genes in the β region. In conclusion, we have identified for the first time two FMCRs of amplification in HGS-EOC, opening up a potential biological role in its etiopathogenesis.     

Insulin-like growth factor (IGF) and hepatocyte growth factor (HGF) in follicular fluid cooperatively promote the oncogenesis of high-grade serous carcinoma from fallopian tube epithelial cells: Dissection of the molecular effects

Incessant ovulation is believed to be a potential cause of epithelial ovarian cancer (EOC). Our previous investigations have shown that insulin-like growth factor (IGF2) and hepatocyte growth factor (HGF) in the ovulatory follicular fluid (FF) contributed to the malignant transformation initiated by p53 mutations. Here we examined the individual and synergistic impacts of IGF2 and HGF on enhancing the malignant properties of high-grade serous carcinoma (HGSC), the most aggressive type of EOC, and its precursor lesion, serous tubal intraepithelial carcinoma (STIC). In a mouse xenograft co-injection model, we observed that FF co-injection induced tumorigenesis of STIC-mimicking cells, FE25. Co-injection with IGF2 or HGF partially recapitulated the tumorigenic effects of FF, but co-injection with both resulted in a higher tumorigenic rate than FF. We analyzed the different transformation phenotypes influenced by these FF growth signals through receptor inhibition. The IGF signal was necessary for clonogenicity, while the HGF signal played a crucial role in the migration and invasion of STIC and HGSC cells. Both signals were necessary for the malignant phenotype of anchoring-independent growth but had little impact on cell proliferation. The downstream signals responsible for these HGF activities were identified as the tyrosine-protein kinase Met (cMET)/mitogen-activated protein kinase and cMET/AKT pathways. Together with the previous finding that the FF-IGF2 could mediate clonogenicity and stemness activities via the IGF-1R/AKT/mammalian target of rapamycin and IGF-1R/AKT/NANOG pathways, respectively, this study demonstrated the cooperation of the FF-sourced IGF and HGF growth signals in the malignant transformation and progression of HGSC through both common and distinct signaling pathways. These findings help develop targeted prevention of HGSC.     

Combination of serum hCG beta and p53 tissue expression defines distinct subgroups of serous ovarian carcinoma

Serous ovarian carcinoma comprises a clinically heterogenous group of tumors, and molecular markers stratifying patients into clinically meaningful subgroups are needed. Numerous markers have been evaluated, but none of them has yet been routinely incorporated into clinical practice. Previously we have found that elevated serum levels of the free beta subunit of human chorionic gonadotropin (hCG beta) and aberrant p53 expression confer poor prognosis in ovarian carcinoma. The aim of our study was to evaluate their combined effect in predicting the outcome of patients with serous ovarian carcinoma. The study material consisted of 173 consecutive patients treated for primary serous ovarian carcinoma in 1 institution between 1990 and 2000. The preoperative serum level of hCG beta was analyzed by a ultrasensitive and specific immunofluorometric assay, and p53 tumor tissue expression by immunohistochemistry using a novel classification. Elevated serum hCG beta (>or=2.0 pmol/L) was detected in 57 (33%) of 173 patients, and aberrant p53 expression in 103 (62%) of 167 interpretable cancers. Elevated hCG beta and aberrant p53 expression were strongly associated with poor prognosis (p < 0.0001 for both). Their additive prognostic effect was marked. Five-year survival was 14% (0-29%) when both markers were aberrant, 44% (29-60%) when either one was aberrant and 82% (70-94%) when both were normal. Preoperative serum hCG beta and tumor tissue p53 expression are feasible markers that divide serous ovarian carcinomas into clinically relevant subgroups.     

Differential and divergent activity of insulin-like growth factor binding protein 6 in platinum-sensitive versus platinum-resistant high-grade serous ovarian carcinoma cell lines

Insulin-like growth factor binding protein 6 (IGFBP6) is a secreted protein with a controversial role in human malignancies, being downregulated in most types of human cancer, but upregulated in selected tumors. Ovarian cancer (OC) is a human malignancy characterized by IGFBP6 downregulation; however, the significance of its low expression during ovarian carcinogenesis is still poorly understood. In the present study, IGFBP6 expression and activation of its associated signaling pathway were evaluated in two matched OC cell lines derived from a high-grade serous OC before and after platinum resistance (PEA1 and PEA2 cells, respectively). A whole genome gene expression analysis was comparatively performed in both cell lines upon IGFBP6 stimulation using Illumina technology. IGFBP6 gene expression data from human OC cases were obtained from public datasets. Gene expression data from public datasets confirmed the downregulation of IGFBP6 in primary and metastatic OC tissues compared with in normal ovarian tissues. The comparative analysis of platinum-sensitive (PEA1) and platinum-resistant (PEA2) cell lines showed quantitative and qualitative differences in the activation of IGFBP6 signaling. Notably, IGFBP6 enhanced ERK1/2 phosphorylation only in PEA1 cells, and induced more evident and significant gene expression reprogramming in PEA1 cells compared with in PEA2 cells. Furthermore, the analysis of selected genes modulated by IGFBP6 (i.e., FOS, JUN, TNF, IL6, IL8 and EGR1) exhibited an inverse regulation in PEA1 versus PEA2 cells. In addition, selected hallmarks (TNFA_signaling_via_NFKB, TGF_beta_signaling, P53_pathway) and IL-6 signaling were positively regulated in PEA1 cells, whereas they were inhibited in PEA2 cells in response to IGFBP6. These data suggested that dysregulation of IGFBP6 signaling may serve a role in the progression of OC, and is likely associated with the development of platinum resistance.     

miR-145在浆液性卵巢癌中的表达及对卵巢癌SKOV3细胞增殖的影响

目的:检测miR-145在浆液性卵巢癌组织及卵巢癌SKOV3细胞中的表达情况,探讨其对卵巢癌SKOV3细胞增殖的影响及其可能的分子机制。方法:利用实时定量PCR技术检测43组浆液性卵巢癌患者癌及癌旁组织中miR-145的表达情况;利用实时定量PCR技术检测正常卵巢浆液腺细胞及SKOV3细胞中miR-145的表达情况;合成miR-145的模拟物miR-145 agomir,将该模拟物及对照分别转染入SKOV3细胞中,应用MTT法检测细胞增殖情况,明确miR-145对卵巢癌SKOV3细胞增殖的影响。结果:实时定量PCR结果显示,与癌旁组织相比,miR-145在浆液性卵巢癌患者癌组织中表达含量降低;与正常细胞相比,miR-145在SKOV3细胞中表达含量降低; MTT结果表明转染了miR-145 angomir的SKOV3细胞增殖受到明显的抑制。结论:miR-145在浆液性卵巢癌组织及SKOV3细胞中均表达含量降低,miR-145可以抑制卵巢癌细胞增殖,miR-145与卵巢癌患者预后密切相关,其含量增高提示好的预后,有望成为新的生物学标志物用于卵巢癌早期诊断及判断预后。     

miR-145 inhibits tumor growth and metastasis by targeting metadherin in high-grade serous ovarian carcinoma

High-grade serous ovarian carcinoma (HGSOC), the most common and aggressive subtype of epithelial ovarian cancer, is characterized by TP53 mutations and genetic instability. Using miRNA profiling analysis, we found that miR-145, a p53 regulated miRNA, was frequently down-regulated in HGSOC. miR-145 down-regulation was further validated in a large cohort of HGSOCs by qPCR. Overexpression of miR-145 in ovarian cancer cells significantly suppressed proliferation, migration and invasion in vitro and inhibited tumor growth and metastasis in vivo. Metadherin (MTDH) was subsequently identified as a direct target of miR-145, and was found to be significantly up-regulated in HGSOC. Furthermore, overexpression of MTDH rescued the inhibitory effects of miR-145 in ovarian cancer cells. Finally, we found that high level of MTDH expression correlated with poor prognosis of HGSOC. Therefore, lack of suppression of MTDH by miR-145 when p53 is dysfunctional leads to increased tumor growth and metastasis of HGSOC. Our study established a new link between p53, miR-145 and MTDH in the regulation of tumor growth and metastasis in HGSOC.     

Circulating cell-free miRNAs as biomarker for triple-negative breast cancer

Background:

Triple-negative breast cancer (TNBC) accounts for 15–20% of all breast cancer in women globally. This subtype often has early and high recurrence rates resulting in poor survival, partially due to lack of targeted therapies. Therefore, there is an urgent need to identify TNBC-specific biomarkers for early diagnosis and treatment monitoring, and to develop more effective targeted therapy.

Methods:

By using miRCURY LNA array platform, we compared the differential miRNA expressions in plasma of patient with TNBC (n=5) and non-TNBC (n=5), as well as healthy controls (n=5). Potential miRNAs were then validated in a large cohort of patients by real-time PCR.

Results:

Ten putative miRNAs from the microarray data that differentially expressed between non-TNBC and healthy controls were identified. In the screening phase (n=90), we selected five miRNAs (miR-92a-3p, miR-342-3p, miR-16, miR-21 and miR-199a-5p) that could discriminate TNBC from non-TNBC for further validation. Results showed that miR-16, miR-21 and miR-199a-5p were underexpressed in TNBC when compared with non-TNBC, and were further validated in a large cohort (n=252). In addition, post-operative plasma levels of miR-16, miR-21 and miR-199a-5p were significantly restored when compared with pre-operative plasma of TNBC. Plasma miR-199a-5p expression in TNBC had significant difference when compared with non-TNBC and healthy controls, the receiver-operator characteristics curve analysis revealed the highest area under curve (AUC=0.8838) among all. The expression levels were associated with TNM stage and tumour subtypes.

Conclusions:

Our data suggest that miR-199a-5p could be a TNBC-specific marker with diagnostic value and provide insights into targeted therapy in the treatment of TNBC.