MiR-146a-5p and miR-191-5p are the Candidates for Novel Diagnostic Markers of Ovarian Clear Cell Carcinoma

Abstract Background: Ovarian cancer is a malignant gynecologic disease rarely diagnosed in the early stages. Among ovarian cancers, clear cell carcinoma has a poor prognosis due to its malignant potential. MicroRNAs (miRNAs) regulate gene expression in cells by suppressing the translation of the target gene or by degrading the target mRNA. They are also secreted from the cells in the blood, binding to the proteins or included in extracellular vesicles lipids and assisting in cell-cell communication. Hence, the serum miRNAs can also be diagnostic biomarkers for ovarian cancer. This study investigated and identified specific miRNAs for ovarian cancer clear cell carcinoma and compared them to those of the ovarian endometrioma in healthy controlpatients. Results: CA125, an ovarian tumor marker, did not differ between ovarian clear cell carcinoma, endometriosis, and healthy controlsthe three groups. Four miRNAs (miR-146a-5p, miR-191-5p, miR-484, and miR-574-3p) were analyzed. The miR-146a-5p and miR-191-5p expression levels were significantly increased in the serum samples from the ovarian clear cell carcinoma subjects compared to the healthy controlscontrols, but not in the subjects with endometriosis (P < 0.05). Furthermore, the bioinformatics analysis showed that CCND2 and NOTCH2 were the candidate target genes of miR‑146a-5p and miR-191-5p.Conclusions: Our results showed that miR‑146a-5p and miR-191-5p might be useful as an early and non-invasive diagnostic tool in ovarian clear cell carcinoma. These miRNAs can help in distinguishing between ovarian clear cell carcinoma and ovarian endometrioma. To the best of our knowledge, no studies have screened any candidates specifically for clear cell carcinoma.

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Frequent PIK3CA mutations in eutopic endometrium of patients with ovarian clear cell carcinoma

Modern Pathology ◽

10.1038/s41379-021-00861-3 ◽

2021 ◽

Author(s):

Kosuke Murakami ◽

Akiko Kanto ◽

Kazuko Sakai ◽

Chiho Miyagawa ◽

Hisamitsu Takaya ◽

...

Keyword(s):

Ovarian Cancer ◽

Cell Carcinoma ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Driver Mutations ◽

Intratumor Heterogeneity ◽

Ovarian Clear Cell Carcinoma ◽

Eutopic Endometrium ◽

Stromal Component ◽

Hotspot Mutations

AbstractRecent studies have reported cancer-associated mutations in normal endometrium. Mutations in eutopic endometrium may lead to endometriosis and endometriosis-associated ovarian cancer. We investigated PIK3CA mutations (PIK3CAm) for three hotspots (E542K, E545K, H1047R) in eutopic endometrium in patients with ovarian cancer and endometriosis from formalin-fixed paraffin-embedded specimens by laser-capture microdissection and droplet digital PCR. The presence of PIK3CAm in eutopic endometrial glands with mutant allele frequency ≥ 15% were as follows: ovarian clear cell carcinoma (OCCC) with PIK3CAm in tumors, 20/300 hotspots in 11/14 cases; OCCC without PIK3CAm, 42/78 hotspots in 11/12 cases; high-grade serous ovarian carcinoma, 8/45 hotspots in 3/5 cases; and endometriotic cysts, 5/63 hotspots in 5/6 cases. These rates were more frequent than in noncancer nonendometriosis controls (7/309 hotspots in 5/17 cases). In OCCC without PIK3CAm, 7/12 (58%) cases showed multiple hotspot mutations in the same eutopic endometrial glands. In 3/54 (5.6%) cases, PIK3CAm was found in eutopic endometrial stroma. Multisampling of the OCCC tumors with PIK3CAm showed intratumor heterogeneity in three of eight cases. In two cases, PIK3CAm was detected in the stromal component of the tumor. Homogenous PIK3CAm in the epithelial component of the tumor matched the mutation in eutopic endometrial glands in only one case. Eutopic endometrial glands in ovarian cancer and endometriosis show high frequency of PIK3CAm that is not consistent with tumors, and multiple hotspot mutations are often found in the same glands. While the mutations identified in eutopic endometrium may not be driver mutations in the patient’s cancer, these are still driver mutations but this specific clone has not undergone the requisite steps for the development of cancer.

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miR-29b regulates cell proliferation and invasion in human ovarian clear cell carcinoma by targeting Lysyl oxidase (LOX)

Archives of Biological Sciences ◽

10.2298/abs150420020w ◽

2016 ◽

Vol 68 (1) ◽

pp. 155-163

Author(s):

Xuan Wang ◽

Yan Wang ◽

Guichan Wang ◽

Peishu Liu

Keyword(s):

Ovarian Cancer ◽

Cell Carcinoma ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Lysyl Oxidase ◽

Gynecologic Cancer ◽

Tumor Formation ◽

Migration And Invasion ◽

Ovarian Clear Cell Carcinoma ◽

Proliferation And Invasion

Ovarian cancer is the leading cause of death from gynecologic cancer, reflecting its chemoresistance and frequent late diagnosis, and suggesting that a more effective treatment approach is needed. Lysyl oxidase (LOX) is involved in important biological processes such as gene regulation, cell signaling and cell motility, its deregulation contributing to tumor formation and development. Although it is known that LOX is involved in proliferation, migration and invasion in several types of tumors, studies of LOX in ovarian cancers are scarce. To explore the molecular regulation mechanisms in ovarian cancer tumorigenesis, the expression change and the function of LOX was confirmed in ovarian tissues and cells, which suggested that LOX is a tumor suppressor gene. To further understand how LOX expression is regulated in ovarian cancer, microRNAs(miRNAs) were considered because of their role in post-transcriptional regulation of many genes. Recent work has described differential expression of mature miRNAs in human cancers. Bioinformatics prediction which was used to find the appropriate miRNA regulating LOX, revealed that miR-29b regulates LOX protein level via its binding site on the 3'UTR of LOX mRNAin ES-2 cells, a human ovarian clear cell carcinoma cell line. miR-29b knockdown inhibited proliferation and invasion in ES-2 cells. Taken together, these findings suggest that influencing LOX regulation bychanging the level of miR-29b expression could provide a novel potential approachfor treating human ovarian clear cell carcinoma.

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Treatment Strategies for ARID1A-Deficient Ovarian Clear Cell Carcinoma

Cancers ◽

10.3390/cancers13081769 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1769

Author(s):

Kazuaki Takahashi ◽

Masataka Takenaka ◽

Aikou Okamoto ◽

David D. L. Bowtell ◽

Takashi Kohno

Keyword(s):

Ovarian Cancer ◽

Cell Carcinoma ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Immune Checkpoint Blockade ◽

Serous Carcinoma ◽

Histological Subtype ◽

Ovarian Clear Cell Carcinoma ◽

Chemotherapeutic Regimen ◽

Platinum Based Chemotherapy

Ovarian clear cell carcinoma (OCCC) is a histological subtype of ovarian cancer that is more frequent in Asian countries (~25% of ovarian cancers) than in US/European countries (less than 10%). OCCC is refractory to conventional platinum-based chemotherapy, which is effective against high-grade serous carcinoma (HGSC), a major histological subtype of ovarian cancer. Notably, deleterious mutations in SWI/SNF chromatin remodeling genes, such as ARID1A, are common in OCCC but rare in HGSC. Because this complex regulates multiple cellular processes, including transcription and DNA repair, molecularly targeted therapies that exploit the consequences of SWI/SNF deficiency may have clinical efficacy against OCCC. Three such strategies have been proposed to date: prioritizing a gemcitabine-based chemotherapeutic regimen, synthetic lethal therapy targeting vulnerabilities conferred by SWI/SNF deficiency, and immune checkpoint blockade therapy that exploits the high mutational burden of ARID1A-deficient tumor. Thus, ARID1A deficiency has potential as a biomarker for precision medicine of ovarian cancer.

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Extracellular miRNAs as Predictive Biomarkers for Glypican-3-Derived Peptide Vaccine Therapy Response in Ovarian Clear Cell Carcinoma

Cancers ◽

10.3390/cancers13030550 ◽

2021 ◽

Vol 13 (3) ◽

pp. 550

Author(s):

Mayu Ukai ◽

Akira Yokoi ◽

Kosuke Yoshida ◽

Shiro Suzuki ◽

Kiyosumi Shibata ◽

...

Keyword(s):

Cell Carcinoma ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Peptide Vaccine ◽

Therapy Response ◽

Predictive Biomarkers ◽

Cancer Tissue ◽

Ovarian Clear Cell Carcinoma ◽

Serum Samples ◽

Extracellular Mirnas

Ovarian clear cell carcinoma (OCCC) has been treated with surgery and chemotherapy; however, the prognosis remains poor because of chemoresistance. Therefore, immunotherapies are attracting attention, including the GPC3 peptide vaccine, which improves overall survival. However, the response rate is limited and there are no sufficient predictive biomarkers that can identify responders before treatment. Our purpose was to identify circulating serum miRNAs as predictive biomarkers for response to GPC3 peptide vaccine. Eighty-four patients in a phase II trial of a GPC3 peptide vaccine were enrolled and miRNA sequencing was performed on their serum samples. Candidate miRNAs were selected from a group of 14 patients for whom treatment was responsive and validated in an independent group of 10 patients for whom treatment was responsive. Three markedly upregulated miRNAs, miR-375-3p, miR-193a-5p, and miR-1228-5p, were identified, and the combination of those miRNAs demonstrated high value in the prediction of the response. The origin of these miRNAs was assessed by referring to OCCC tissue miRNA profiles, and they were not identified as cancer tissue-related miRNAs. Functional annotation analysis suggested that they were associated with interferon-related pathways. The miRNAs identified herein have great potential to allow the realization of liquid biopsy for predicting the immunotherapy response and precision medicine.

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G Protein–Coupled Receptor 30 Mediates the Anticancer Effects Induced by Eicosapentaenoic Acid in Ovarian Cancer Cells

Cancer Research and Treatment ◽

10.4143/crt.2019.380 ◽

2020 ◽

Vol 52 (3) ◽

pp. 815-829 ◽

Cited By ~ 1

Author(s):

Yue Zhao ◽

Meng-Fei Zhao ◽

Mei-Lin Yang ◽

Tian-Yu Wu ◽

Cong-Jian Xu ◽

...

Keyword(s):

Ovarian Cancer ◽

Fatty Acids ◽

Cell Carcinoma ◽

G Protein ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Carcinoma Cells ◽

Omega 3 ◽

Ovarian Clear Cell Carcinoma ◽

G Protein Coupled

PurposeWhile numerous epidemiological studies have indicated that omega-3 polyunsaturated fatty acids have anticancer properties in various cancers, the effects and mechanisms of eicosapentaenoic acid (EPA) in ovarian cancer cell growth are poorly understood.Materials and MethodsES2 ovarian clear cell carcinoma cells and SKOV3 adenocarcinoma cells were treated with palmitic acid or EPA, followed by flow cytometry and cell counting to measure apoptosis and proliferation, respectively. A modified protein lipid overlay assay was used to further verify whether EPA was a ligand of G protein–coupled receptor 30 (GPR30) in ES2 cells. The levels of apoptosis-related genes, phosphorylated AKT, and phosphorylated ERK1/2 were detected to explore the underlying mechanism. Finally, inhibitory effect of EPA on tumor growth via GPR30 was determined <i>in vitro</i> and <i>in vivo</i>.ResultsEPA suppressed ES2 ovarian clear cell carcinoma cells growth via GPR30, a novel EPA receptor, by inducing apoptosis. As a ligand of GPR30, EPA activated the GPR30-cAMP– protein kinase A signaling pathway. When GPR30 was suppressed by siRNA or its inhibitor G15, the antiproliferative action of EPA was impaired. Furthermore, EPA inhibited tumor growth by blocking the activation of AKT and ERK. In the mouse xenograft model, EPA decreased tumor volume and weight through GPR30 by blocking tumor cell proliferation.ConclusionThese results confirm that EPA is a tumor suppressor in human ovarian clear cell carcinoma cells and functions through a novel fatty acid receptor, GPR30, indicating a mechanistic linkage between omega-3 fatty acids and cancers.

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Frequent PIK3CA mutations in eutopic endometrium of patients with ovarian clear cell carcinoma

10.1101/2021.02.25.432943 ◽

2021 ◽

Author(s):

Kosuke Murakami ◽

Akiko Kanto ◽

Kazuko Sakai ◽

Chiho Miyagawa ◽

Hisamitsu Takaya ◽

...

Keyword(s):

Ovarian Cancer ◽

Cell Carcinoma ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Driver Mutations ◽

Intratumor Heterogeneity ◽

Ovarian Clear Cell Carcinoma ◽

Eutopic Endometrium ◽

Stromal Component ◽

Hotspot Mutations

AbstractRecent studies have reported cancer-associated mutations in normal endometrium. Mutations in eutopic endometrium may lead to endometriosis and endometriosis-associated ovarian cancer. We investigated PIK3CA mutations (PIK3CAm) for three hotspots (E542K, E545K, H1047R) in eutopic endometrium in patients with ovarian cancer and endometriosis from formalin-fixed paraffin-embedded specimens by laser-capture microdissection and droplet digital PCR. The presence of PIK3CAm in eutopic endometrial glands with mutant allele frequency ≥15% were as follows: ovarian clear cell carcinoma (OCCC) with PIK3CAm in tumors, 20/300 hotspots in 11/14 cases; OCCC without PIK3CAm, 42/78 hotspots in 11/12 cases; high-grade serous ovarian carcinoma, 8/45 hotspots in 3/5 cases; and endometriotic cysts, 5/63 hotspots in 5/6 cases. These rates were more frequent than in non-cancer non-endometriosis controls (7/309 hotspots in 5/17 cases). In OCCC without PIK3CAm, 7/12 (58%) cases showed multiple hotspot mutations in the same eutopic endometrial glands. In 3/54 (5.6%) cases, PIK3CAm was found in eutopic endometrial stroma. Multi-sampling of the OCCC tumors with PIK3CAm showed intratumor heterogeneity in three of eight cases. In two cases, PIK3CAm was detected in the stromal component of the tumor. Homogenous PIK3CAm in the epithelial component of the tumor matched the mutation in eutopic endometrial glands in only one case. Eutopic endometrial glands in ovarian cancer and endometriosis show high frequency of PIK3CAm that is not consistent with tumors, and multiple hotspot mutations are often found in the same glands. Our results suggest that most PIK3CAm in eutopic endometrial glands are passenger rather than driver mutations.

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Human monoclonal antibody for ovarian clear cell carcinoma-2, a human monoclonal antibody with antitumor activity against ovarian cancer cells that recognizes CA125-like antigen

International Journal of Gynecological Cancer ◽

10.1111/j.1525-1438.2007.01147.x ◽

2008 ◽

Vol 18 (5) ◽

pp. 996-1006

Author(s):

N. Suzuki ◽

Y. Tamada ◽

K. Shigirahara ◽

A. Suzuki ◽

N. Susumu ◽

...

Keyword(s):

Ovarian Cancer ◽

Monoclonal Antibody ◽

Antitumor Activity ◽

Cell Carcinoma ◽

Cancer Cells ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Human Monoclonal Antibody ◽

Ovarian Cancer Cells ◽

Ovarian Clear Cell Carcinoma

In recent years, antibody therapy employing monoclonal antibodies has become a new approach for treating cancer. This study was performed to establish a human monoclonal antibody recognizing an epitope related to CA125 using KM mice and to assess its reactivity with ovarian cancer cells. A human ovarian clear cell adenocarcinoma cell line (RMG-I) was used to immunize KM mice, and hybridoma supernatant was obtained by a standard method employing enzyme-linked immunosorbent assay screening. Next, selection of hybridomas was performed with two antibodies (MA602-1 and MA602-6) and a sandwich immunoassay for CA125-like antigen, and then the limiting dilution was used to obtain a human monoclonal antibody. Immunohistochemical reactivity of this antibody (human monoclonal antibody for ovarian clear cell carcinoma-2 [HMOCC-2]) with ovarian cancer was assessed, while its specificity was analyzed by Western blotting. Various antibodies were used to identify the epitope targeted by HMOCC-2. Finally, the antitumor effect of HMOCC-2 was assessed by intraperitoneal administration to SCID (severe combined immunodeficiency) mice with heterografts of RMG-I tumors. HMOCC-2 showed a positive reaction with 60% (63/105) of ovarian cancer specimens. Western blotting of the membrane fraction of RMG-I revealed several bands at 120–250 kd. HMOCC-2 recognized the CA125-like antigens identified by several antibodies. HMOCC-2 also exhibited significant antitumor activity (P< 0.01) against ovarian cancer heterografts. HMOCC-2 reacts specifically with ovarian cancer cells via a target epitope analogous to that of CA125 and also exhibits activity against ovarian tumors. These findings suggest that it may have the potential to be employed clinically for molecular-targeting therapy.

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GLS1 is a protective factor rather than a molecular target in ARID1A-mutated ovarian clear cell carcinoma

10.1101/2021.09.03.457161 ◽

2021 ◽

Author(s):

Valentino Clemente ◽

Andrew Nelson ◽

Britt Erickson ◽

Ruth Baker ◽

Nathan Rubin ◽

...

Keyword(s):

Ovarian Cancer ◽

Cell Carcinoma ◽

Protective Factor ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Glutamine Metabolism ◽

Ovarian Clear Cell Carcinoma ◽

Human Cancers ◽

Key Enzyme

AbstractTargeting glutamine metabolism has emerged as a novel therapeutic strategy for several human cancers, including ovarian cancer. The primary target of this approach is the kidney isoform of glutaminase, glutaminase 1 (GLS1), a key enzyme in glutamine metabolism that is overexpressed in several human cancers. A first-in-class inhibitor of GLS1, called CB839 (Telaglenastat), has been investigated in several clinical trials, with promising results. The first clinical trial of CB839 in platinum-resistant ovarian cancer patients is forthcoming. ARID1A-mutated ovarian clear cell carcinoma (OCCC) is a relatively indolent and chemoresistant ovarian cancer histotype. In OCCC-derived cells in vitro and mouse models, loss of ARID1A leads to upregulation of GLS1. Thus, targeting of GLS1 with CB839 has been suggested as a targeted approach for OCCC patients with tumors harboring ARID1A-mutations. Here, we investigated whether GLS1 is differentially expressed between OCCC patients whose tumors are ARID1A positive and patients whose tumors are ARID1A negative. In clinical specimens of OCCC, we found that GLS1 overexpression was not correlated with ARID1A loss. In addition, GLS1 overexpression was associated with better clinical outcomes. Our findings suggest that GLS1 expression in OCCC may be a protective factor and that caution should be taken when considering the use of CB839 to treat OCCC patients.

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