Heterogeneous genetic alterations in ovarian mucinous tumors: Application and usefulness of laser capture microdissection

Takeshima, Yukio; Amatya, Vishwa Jeet; Daimaru, Yutaka; Nakayori, Fumio; Nakano, Tomohiro; Inai, Kei

doi:10.1053/hupa.2001.28956

Cited by 16 publications

(8 citation statements)

References 20 publications

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“…80 Disparities in DNA content and K-Ras mutations have been noted between primary ovarian tumors and corresponding lymph-node and peritoneal metastases, further supporting the importance of ITH. 27,81,82 Significant differences have also been observed in immunohistochemical staining of mismatch repair proteins, including high mMSH2 staining in metastatic versus primary tumors. Additional evidence comes from Khalique et al who confirmed ITH in 16 cases of grade 3 serous EOCs using microsatellite and single nucleotide polymorphism (SNP) analyses.…”

Section: Discussionmentioning

confidence: 93%

Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform

et al. 2013

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The use of biomarkers to infer drug response in patients is being actively pursued, yet significant challenges with this approach, including the complicated interconnection of pathways, have limited its application. Direct empirical testing of tumor sensitivity would arguably provide a more reliable predictive value, although it has garnered little attention largely due to the technical difficulties associated with this approach. We hypothesize that the application of recently developed microtechnologies, coupled to more complex 3-dimensional cell cultures, could provide a model to address some of these issues. As a proof of concept, we developed a microfluidic device where spheroids of the serous epithelial ovarian cancer cell line TOV112D are entrapped and assayed for their chemoresponse to carboplatin and paclitaxel, two therapeutic agents routinely used for the treatment of ovarian cancer. In order to index the chemoresponse, we analyzed the spatiotemporal evolution of the mortality fraction, as judged by vital dyes and confocal microscopy, within spheroids subjected to different drug concentrations and treatment durations inside the microfluidic device. To reflect microenvironment effects, we tested the effect of exogenous extracellular matrix and serum supplementation during spheroid formation on their chemotherapeutic response. Spheroids displayed augmented chemoresistance in comparison to monolayer culturing. This resistance was further increased by the simultaneous presence of both extracellular matrix and high serum concentration during spheroid formation. Following exposure to chemotherapeutics, cell death profiles were not uniform throughout the spheroid. The highest cell death fraction was found at the center of the spheroid and the lowest at the periphery. Collectively, the results demonstrate the validity of the approach, and provide the basis for further investigation of chemotherapeutic responses in ovarian cancer using microfluidics technology. In the future, such microdevices could provide the framework to assay drug sensitivity in a timeframe suitable for clinical decision making.

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Section: Discussionmentioning

confidence: 93%

Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform

et al. 2013

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“…The DNA was extracted from these microdissected tissues as per the previously described method 12 . To avoid contamination of the tumor tissues and stromal tissues, these components were also microdissected using Laser Capture Microdissection (LCM; LM200; Arcturus Engineering, Santa Clara, CA, USA) and DNA was extracted as previously reported 13 . The analysis of TP53 point mutation (exons 5–8) was performed in extracted DNA using ABI Prism 310 Genetic Analyzer (Perkin‐Elmer Japan Co., Chiba, Japan) according to the manufacturer's directions.…”

Section: Case Reportmentioning

confidence: 99%

Esophageal carcinosarcoma with basaloid squamous carcinoma and rhabdomyosarcoma components with TP53 mutation

Amatya

Takeshima

Kaneko

et al. 2004

Pathology International

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Carcinosarcoma of the esophagus is a rare tumor with a distinct pathological entity having squamous cell carcinoma as the most described carcinomatous component. This paper reports the first case of carcinosarcoma of the esophagus that showed predominant basaloid squamous carcinoma component in addition to squamous cell carcinoma and poorly differentiated carcinoma and sarcoma component. A 64-year-old male patient consulted for dysphasia and chest pain was examined and found to have gastrointestinal fiber-endoscope and a polypoid growth in the lower third of the esophagus. Partial esophagectomy was performed and the excised tumor showed histological features of carcinosarcoma with heterogeneous carcinomatous components with dominance of basaloid squamous carcinoma and minority of squamous cell carcinoma, poorly differentiated carcinoma, and sarcomatous component, immunohistochemically proven to be rhabdomyosarcoma. Immunohistochemical study and TP53 mutation analysis was carried out to explain the histogenesis of this rare tumor. The distinct immunohistochemical profiles of the carcinomatous and sarcomatous components suggested the possibility of transition from a carcinomatous to a sarcomatous component. The similar TP53 mutation in the carcinomatous and sarcomatous component suggested each of these components had the same origin, that is, the tumor was monoclonal in origin.

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“…DNA, RNA or protein analyses may be performed with the microdissected tissue by any method with adequate sensitivity. For example, DNA extracted from cells procured via this technique can be used for loss-of-heterozygosity analysis [63][64][65][66][67], DNA methylation assays [62], production of cDNA libraries [68,69], gene expression assays [25,70,71], RT-PCR and quantitative PCR (QPCR) [49,72,73], while protein extracted from microdissected [75][76][77] and mass spectrometry [78][79][80][81][82][83][84].…”

Section: Diversity Of Applicationsmentioning

confidence: 99%

Laser capture microdissection technology

Espina

Heiby

Pierobon

et al. 2007

Expert Review of Molecular Diagnostics

166

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Deciphering the cellular and molecular interactions that drive disease within the tissue microenvironment holds promise for discovering drug targets of the future. In order to recapitulate the in vivo interactions through molecular analysis, one must be able to analyze specific cell populations within the context of their heterogeneous tissue microecology. Laser capture microdissection is a method to procure subpopulations of tissue cells under direct microscopic visualization. Laser capture microdissection technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity analysis, RNA transcript profiling, cDNA library generation, mass spectrometry proteomics discovery and signal pathway profiling.

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Heterogeneous genetic alterations in ovarian mucinous tumors: Application and usefulness of laser capture microdissection

Cited by 16 publications

References 20 publications

Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform

Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform

Esophageal carcinosarcoma with basaloid squamous carcinoma and rhabdomyosarcoma components with TP53 mutation

Laser capture microdissection technology

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