Development Of Clear Cell Carcinoma Research Articles

Clear cell carcinoma of the ovary: Epidemiology, pathological and biological features, treatment options and clinical outcomes

Clear cell carcinoma of the ovary is a rare and distinct histotype of epithelial ovarian carcinomas. Women diagnosed with clear cell carcinomas are usually younger and diagnosed at earlier stages than those with the most common high-grade serous histology. Endometriosis is considered a main risk factor for the development of clear cell carcinoma of the ovary, and it can be considered a precursor of of this tumor, as it is identified in more than 50% of patients with clear cell carcinoma.Different molecular pathways and alterations heve been identified in ovarian clear cell carcinoma, including the most common mutations of AT-rich interaction domain 1A [ARID1A] and phosphatidylinositol-4,5-bisphosphate 3-kinase [PIK3] catalytic subunit alpha [PIK3CA].The prognosis of patients at early stage is favorable, while patients with advanced or recurrent disease experience a poor oncologic outcomes.Despite a lower rate of responses due to an intrinsic chemoresistance, the treatment strategy for advanced disease resembles the treatment of high-grade serous carcinoma, which includes aggressive cytoreductive surgery and platinum-based chemotherapy. For this reason, the role of adjuvant chemotherapy in patients with stage I disease undergoing complete surgical staging is still under debate.Alternative treatments, including biological agents that target different pathways constitute the most promising treatment strategies, and well-designed, collaborative international trials should be designed in order to improve the oncologic outcomes and the quality of life of patients with this aggressive disease.

Delphinidin suppresses proliferation and migration of human ovarian clear cell carcinoma cells through blocking AKT and ERK1/2 MAPK signaling pathways

Delphinidin possesses the highest chemopreventive activity among the six components of anthocyanidin that are pigments from fruits and vegetables giving them blue, purple or red colors. Although delphinidin has anti-carcinogenic and apoptotic effects in various cancers, little is known about its functional roles in ovarian clear cell carcinoma (CCC) which shows poor prognosis with resistance to chemotherapy as compared with other subtypes of epithelial ovarian cancers (EOC). Results of present study revealed that cell survival rates of ES2 cells from ovarian CCC treated with delphinidin decreased in a dose-dependent manner. Also, delphinidin inhibited migration and induced apoptosis of ES2 cells. To investigate the molecular mechanisms responsible for biological effects of delphinidin, we analyzed the phosphorylation status of carcinogenic protein kinases related to development of CCC in a dose- and time-dependent manner. Phosphorylation of downstream targets of PI3K (AKT and p70S6K) and MAPKs (ERK1/2 and JNK) signaling was suppressed by treatment of ES2 cells with delphinidin. In addition, pharmacological inhibitors of PI3K/AKT and ERK1/2 MAPK pathway improved the anti-proliferative action of delphinidin on ES2 cells. Moreover, we compared the cancer preventive effects of delphinidin with traditional cisplatin- and paclitaxel-based chemotherapy on cell viability of ES2 cells. Results showed that delphinidin is as effective in its therapeutic activity against ES2 cells as cisplatin and placlitaxel. Collectively, these results indicated that delphinidin plays a critical role as a new chemotherapeutic agent to prevent development and progression of ES2 cells in CCC via inactivation of PI3K/AKT and ERK1/2 MAPK signal transduction cascades.

Identifying potential therapeutics by molecular profiling of 136 cases of uterine clear cell carcinoma

• Uterine clear cell carcinoma is a rare subtype of endometrial cancer that accounts for only 1-6% of all uterine malignancies.1-2 • Uterine clear cell carcinomas are considered to be Type-II estrogen-independent and exhibit aggressive behavior with worse clinical outcomes compared Type I tumors.3 • Uterine clear cell carcinomas most often occur in post menopausal women with a median age of 66-68 years.4 • Similar to uterine serous cancers, clear cell carcinoma is more prevalent in African American women.5 • While the pathogenesis of many subtypes of uterine cancer have been well studied, the molecular pathways involved in uterine clear cell carcinoma development are poorly understood. • Broader understanding of the molecular and genomic characteristics of uterine clear cell carcinomas could contribute to the growing body of literature this disease and potentially provide treatment options with targeted therapies. Background: Clear cell carcinoma of the endometrium (CCE) is a rare subtype of endometrial cancer (EC) associated with worse prognosis when compared to other high grade EC subtypes. We aim to evaluate molecular, genomic and protein expression patterns in a large cohort of CCEs in order to direct patients to rational therapeutic strategies and clinical trials. Methods: Out of 3133 EC submitted to Caris Life Sciences from March 2011 to July 2014, 136 CCEs were identified based on reported pathology. Testing was ordered per physician request and included a combination of sequencing (Sanger or next generation sequencing), protein expression (immunohistochemistry), and /or gene amplification (FISH/CISH). Results: Of the samples evaluated by sequencing, the most common genetic mutations were P53 (40%) and BRCA2 (33%). Hormone receptor expression was low: ERα (35%), PR (22%) and AR (7%). ERBB2 was mutated in 8%, while its protein product Her2, a biomarker for HER2-directed therapies, was amplified in 12% and expressed in 5% of patients. Aberrations of the PI3K pathway, including 26% PTEN and 25% PIK3CA mutation rate, with 69% loss of PTEN expression, highlight potential utility with inhibitors of this pathway. DNA repair pathway was altered in our CCE cohort as well: low ERCC1 (6%) and MGMT (34%) expression, and high BRCA2 mutation rate, suggesting sensitivity to alkylating agents. Evaluation of the cMET pathway showed 40% IHC expression. TUBB3, a class III β-tubulin, was infrequently expressed (15%), and TLE3 expression was high (29%) compared to other EC subtypes, implicating sensitivity to microtubule-stabilizing agents. Increased TOPO2A expression, associated with anthracycline efficacy, was seen in over 80% of cases. Loss of RRM1, a DNA synthesis protein known to determine efficacy of gemcitabine, was seen in 78% of cases. Conclusions: Our findings highlight the genetic heterogeneity of CCE, and identified altered cellular pathways with potential diagnostic and predictive values for therapeutic intervention. Drugs targeting the pathways for DNA repair, PI3K, and receptor tyrosine kinases, as well as gemcitabine and taxanes, may warrant consideration in selected patients with CCE. • 136 out of 3133 (4.3%) of endometrial cancers submitted to Caris Life Sciences from March 2011 to July 2014 were identified as clear cell carcinoma. • Specific testing was performed per physician request and included a combination of sequencing (Sanger, NGS or pyrosequencing), protein expression (IHC), gene amplification (CISH or FISH), and/or RNA fragment analysis. • IHC analysis was performed on formalin-fixed paraffin-embedded tumor samples using commercially available detection kits, automated staining techniques (Benchmark XT, Ventana, and AutostainerLink 48, Dako), and commercially available antibodies. • Fluorescent in-situ hybridization (FISH) was used for evaluation of the HER-2/neu [HER-2/CEP17 probe], EGFR [EGFR/CEP7 probe], and cMET [cMET/CEP7 probe] (Abbott Molecular/Vysis). HER-2/neu and cMET status were also evaluated by chromogenic in-situ hybridization (INFORM HER-2 Dual ISH DNA Probe Cocktail; commercially available cMET and chromosome 7 DIG probe; Ventana). The same scoring system was applied as for FISH. • Direct sequence analysis was performed on genomic DNA isolated from formalin-fixed paraffin-embedded tumor samples using the Illumina MiSeq platform. Specific regions of 47 genes of the genome were amplified using the Illumina TruSeq Amplicon Cancer Hotspot panel. • Mutation analysis by Sanger sequencing included selected regions of BRAF, KRAS, NRAS, c-KIT, EGFR, and PIK3CA genes and was performed by using M13-linked PCR primers designed to amplify targeted sequences. • Retrospective data analysis; Statistical analysis (unpaired t-tests used to compare biomarker expression across histologic subtypes) performed using PrismTM v6. Biomarker associations were calculated by two-tailed Fisher Exact tests.

Are oestrogens and genetic predisposition etiologic factors in the development of clear cell carcinoma of the peritoneum?

A literature search was carried out for clinical observations that could explain the possible aetiology of primary peritoneal clear cell carcinoma (CCC) including diagnostic dilemmas, various theories of origin, oestrogen dependence and genetic association. It was found to be an extremely rare tumour (CCC) arising in the peritoneum and is often misdiagnosed as mesothelioma or serous carcinoma or metastatic adenocarcinoma due to overlapping morphological features. The awareness of such dilemmas is important even before making a diagnosis. Clinicopathological features and immunohistochemical studies like WT1, CK20 and calretinin are usually helpful in differentiating CCC from serous carcinoma, metastatic carcinoma from bowel and mesothelioma. (CK7 is common to all epithelial tumours, CEA can be expressed in clear cell carcinoma, WT1 is normally expressed in serous carcinoma, calretinin is expressed in mesothelioma and CK20 in colon carcinoma). The distinction between the above tumours is important as correct diagnosis is required in initiating appropriate treatment.Less than ten cases have been reported in the English language medical literature. Mullerian metaplasia and malignant transformation in endometriosis are two theories proposed for its existence. Peritoneal CCC can be oestrogen dependent in view of association with oestrogen dependent conditions which include endometriosis, adenomyosis and endometrial carcinoma. A genetic ascertain is difficult to be associated and needs further evaluation in a research setting in the familial cancer genetic clinics.

Abstract 1117: Macrophage migration inhibitory factor (MIF) plays a critical role in clear cell renal carcinoma

Abstract Clear cell renal carcinoma (CCRC) is the most common form of renal cancer, resulting in 50,000 new cases per year in the US, and greater than 10,000 deaths. CCRC is frequently characterized by alterations in the von Hippel-Lindau protein (VHL), an E3 ligase that mediates oxygen dependent degradation of the alpha subunit of the hypoxia inducible factor (HIF). Activation of HIF and many of its transcriptional target genes are critical for renal tumorigenesis. The macrophage migration inhibitor factor (MIF) is a pleiotropic cytokine that has roles in inflammation, cell signaling, and cell cycle regulation. MIF is regulated by HIF, and is expressed in a variety of tumor types wherein MIF expression correlates with poor outcome. Functionally, MIF can negatively regulate p53, and thus may be particularly relevant in cancers like clear cell carcinoma in which p53 is not commonly mutated. The purpose if this study is to determine the role of MIF in CCRC. In order to determine if MIF is expressed in CCRC patient samples, we stained tumor microarrays with over 95 primary CCRC tumors for MIF expression. In keeping with its reported regulation by the VHL/HIF pathway, MIF demonstrated high expression in greater than 90% of the tumors compared to normal kidney samples. Likewise, in CCRC tumor cell lines MIF expression was highly expressed compared to cell lines in which VHL expression has been reconstituted. To determine the significance of MIF expression in CCRC tumor cell lines, MIF was depleted using two independent shRNAs and we observed significant decreases cell proliferation and clonogenic survival in a variety of CCRC cell lines. These observations were replicated by silencing the expression of the MIF receptor CD74, suggesting the MIF signaling pathway is intact and functional CCRC. At the mechanistic level, MIF is known to inhibit the function of p53 through promotion of MDM2 mediated degradation. To assess whether the role of MIF in CCRC is to regulate p53, we determined the effects on p53 signaling when MIF expression was inhibited. We found that inhibition of MIF lead to an increase in p53 stability and subsequent expression of the canonical p53 target gene p21. Together these observations suggest that MIF expression may play a critical role as a target of the HIF/VHL pathway that leads to the development of clear cell carcinoma. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1117. doi:10.1158/1538-7445.AM2011-1117

Role of everolimus in the treatment of renal cell carcinoma

The therapeutic options in metastatic renal cell carcinoma have been recently expanded by the discovery of the VHL gene, the mutation of which is associated with development of clear cell carcinoma, and overexpression of the angiogenesis pathway, resulting in a very vascular tumor. This breakthrough in science led to the development of a variety of small molecules inhibiting the VEGF-dependent angiogenic pathway, such as sunitinib and sorafenib. These agents prolong overall and progression-free survival, respectively. The result was the development of robust front-line therapies which ultimately fail and are associated with disease progression. In this setting, there existed an unmet need for developing second-line therapies for patients with refractory metastatic renal cell carcinoma (MRCC). Everolimus (RAD 001) is an oral inhibitor of the mammalian target of rapamycin (mTOR) pathway. The double-blind, randomized, placebo-controlled phase III trial of everolimus (RECORD-1) conducted in MRCC patients after progression on sunitinib or sorafenib, or both, demonstrated a progression-free survival benefit favoring the study drug (4.9 months vs 1.9 months, HR 0.33, 95% CI 0.25 to 0.43, P </= 0 0.001). Everolimus thus established itself as a standard of care in the second-line setting for patients with MRCC who have failed treatment with VEGF receptor inhibitors.

The Role of Hepatocyte Nuclear Factor-1β in the Pathogenesis of Clear Cell Carcinoma of the Ovary

Clear cell carcinoma (CCC) of the ovary has a number of features distinguishing it from other epithelial ovarian carcinomas (EOC) because of its characteristic histology and biology, frequent concurrence with endometriotic lesion, and highly chemoresistant nature resulting in an extremely poor prognosis. The incidence of CCC has been steadily increasing in Japan. They comprise approximately 20% of all EOC. Understanding the mechanisms of CCC development and elucidating pathogenesis and pathophysiology are intrinsic to prevention and effective therapies for CCC. This article reviews the English language literature for biology, pathogenesis, and pathophysiological studies on endometriosis-associated EOC. Several data are discussed in the context of endometriosis and CCC biology. Recent studies based on genome-wide expression analysis technology have noted specific expression of hepatocyte nuclear factor-1beta (HNF-1beta) in endometriosis and CCC, suggesting that early differentiation into the clear cell lineage takes place in the endometriosis. The HNF-1beta-dependent pathway of CCC will be discussed, which are providing new insights into regulation of apoptosis and glycogen synthesis and resistance of CCC to anticancer agents. This review summarizes recent advances in the HNF-1beta and its target genes; the potential challenges to the understanding of carcinogenesis, pathogenesis, and pathophysiology of CCC; and a possible novel model is proposed.

P53 Mutation Is Infrequent in Clear Cell Carcinoma of the Ovary

Objective. p53 gene alteration has been extensively studied in epithelial ovarian cancer. However, its occurrence in clear cell carcinoma, an infrequent histologic subtype of epithelial ovarian cancer, is rarely reported. The aim of this study is to determine the status of p53 gene alteration in this distinct type of ovarian carcinoma.Methods. Paraffin blocks of tumors from 38 patients with primary or recurrent ovarian clear cell carcinoma were studied for p53 alteration. All these tumors were subjected to immunohistochemical and molecular analysis. Two monoclonal antibodies (DO-7 and PAb 1801) were used for immunohistochemical staining. Genomic DNAs extracted from paraffin blocks of the 38 tumors were subscribed for a nested polymerase chain reaction/single-strand conformation polymorphism (PCR/SSCP) analysis. Tumors showing band shift on SSCP were further prepared for DNA sequencing to determine the site of mutation.Results. Overexpression of p53 was observed in only one stage III clear cell carcinoma. However, focal positive p53 staining was noted in another five tumors. Of the six tumors showing positive immunohistochemistry, p53 alterations were noted in four tumors. Three tumors revealed a missense point mutation: two were in exon 7 (TCT227 → TTT and GGC245 → AGC) and one was in exon 5 (CGC156 → CAC). Another tumor revealed a 12-bp deletion in two possible ways: it might involve the last four codons at the 3′ end of exon 4 (nucleotides 12,288–12,299) or it might cross over the splice junction between exon 4 and intron 4 (nucleotides 12,290–12,301). The former would result in a predicted protein product of 389 amino acids whereas the latter would cause a frameshift in the gene sequence and would result in a truncated protein.Conclusion. Mutations in p53 appear to be much less frequent in clear cell carcinoma than in other histologic types of epithelial ovarian cancer. We suggest that p53 alterations may not play an important role in the development of clear cell carcinoma.

Clear cell carcinoma of the endometrium is characterized by a distinctive profile of p53, Ki-67, estrogen, and progesterone receptor expression

This study was designed to analyze certain clinicopathological features and the profile of p53, Ki-67, estrogen (ER), and progesterone (PR) receptor expression of clear cell carcinoma of the endometrium and to determine whether the pathogenesis of clear cell carcinoma can be accommodated by a dualistic model of endometrial carcinogenesis. In this model, endometrioid carcinoma develops from endometrial hyperplasia under unopposed estrogenic stimulation, and serous carcinoma develops in atrophic endometrium from a putative precursor lesion designated endometrial intraepithelial carcinoma (EIC). Twenty-one clear cell carcinomas of the endometrium were analyzed and compared with 77 endometrioid carcinomas of all grades and 30 serous carcinomas. Clear cell carcinomas showed a distinctive immunoprofile characterized by immunonegativity for ER and PR, low immunoreactivity for p53, and a high Ki-67 proliferation index. ER, PR, and Ki-67 expression were similar to serous carcinoma, but p53 expression was significantly lower in clear cell carcinoma (P < .05). ER and PR expression were significantly lower, and the Ki-67 proliferation index was significantly higher in clear cell carcinoma compared with endometrioid carcinomas (P < .05). p53 expression tended to be higher in clear cell carcinoma compared with endometrioid carcinoma, but the difference was not statistically significant. In contrast to endometrioid carcinoma, clear cell carcinoma was rarely associated with endometrial hyperplasia and serous carcinoma was not. Subdividing clear cell carcinoma morphologically into one that resembled serous carcinoma (clear cell carcinoma with serous features) and another that did not (typical clear cell carcinoma) showed that clear cell carcinoma with serous features had a higher Ki-67 proliferation index than typical clear cell carcinoma, although expression of ER, PR, and p53 were similar. Clear cell carcinoma with serous features was associated with EIC in 50% and was not associated with endometrial hyperplasia. In contrast, typical clear cell carcinoma was associated with endometrial hyperplasia in 40% and was not associated with EIC. In summary, this study provides evidence that clear cell carcinoma of the endometrium, like serous carcinoma, is estrogen independent and shows a high Ki-67 proliferation index. In contrast to serous carcinoma, strong p53 expression occurred less frequently in clear cell carcinoma and predominantly in clear cell carcinoma with serous features. The findings suggest that the molecular events that underlie the development of clear cell carcinoma differ from those of endometrioid and serous carcinoma.