NDLI: Generation and Characterisation of a Canine EGFP-HMGA2 Prostate Cancer In Vitro Model

Content Provider	PubMed Central
Author	Willenbrock, Saskia Wagner, Siegfried Nicola, Reimann-berg Moulay, Md Marion, Hewicker-trautwein Nolte, Ingo Escobar, Hugo Murua
Editor	Culig, Zoran
Copyright Year	2014
Abstract	The architectural transcription factor HMGA2 is abundantly expressed during embryonic development. In several malignant neoplasias including prostate cancer, high re-expression of HMGA2 is correlated with malignancy and poor prognosis. The let-7 miRNA family is described to regulate HMGA2 negatively. The balance of let-7 and HMGA2 is discussed to play a major role in tumour aetiology. To further analyse the role of HMGA2 in prostate cancer a stable and highly reproducible in vitro model system is precondition. Herein we established a canine CT1258-EGFP-HMGA2 prostate cancer cell line stably overexpressing HMGA2 linked to EGFP and in addition the reference cell line CT1258-EGFP expressing solely EGFP to exclude EGFP-induced effects. Both recombinant cell lines were characterised by fluorescence microscopy, flow cytometry and immunocytochemistry. The proliferative effect of ectopically overexpressed HMGA2 was determined via BrdU assays. Comparative karyotyping of the derived and the initial CT1258 cell lines was performed to analyse chromosome consistency. The impact of the ectopic HMGA2 expression on its regulator let-7a was analysed by quantitative real-time PCR. Fluorescence microscopy and immunocytochemistry detected successful expression of the EGFP-HMGA2 fusion protein exclusively accumulating in the nucleus. Gene expression analyses confirmed HMGA2 overexpression in CT1258-EGFP-HMGA2 in comparison to CT1258-EGFP and native cells. Significantly higher let-7a expression levels were found in CT1258-EGFP-HMGA2 and CT1258-EGFP. The BrdU assays detected an increased proliferation of CT1258-HMGA2-EGFP cells compared to CT1258-EGFP and native CT1258. The cytogenetic analyses of CT1258-EGFP and CT1258-EGFP-HMGA2 resulted in a comparable hyperdiploid karyotype as described for native CT1258 cells. To further investigate the impact of recombinant overexpressed HMGA2 on CT1258 cells, other selected targets described to underlie HMGA2 regulation were screened in addition. The new fluorescent CT1258-EGFP-HMGA2 cell line is a stable tool enabling in vitro and in vivo analyses of the HMGA2-mediated effects on cells and the development and pathogenesis of prostate cancer.
Related Links	http://dx.doi.org/10.1371/journal.pone.0098788
Starting Page	98788
File Format	PDF
ISSN	19326203
e-ISSN	19326203
Journal	PLoS ONE
Issue Number	6
Volume Number	9
Language	English
Publisher	Public Library of Science
Publisher Date	2014-06-10
Access Restriction	Open
Rights Holder	Public Library of Science
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

CircTHSD4 promotes the malignancy and docetaxel (DTX) resistance in prostate cancer by regulating miR-203/HMGA2 axis

Generation of a syngeneic orthotopic transplant model of prostate cancer metastasis

RNA-seq of nine canine prostate cancer cell lines reveals diverse therapeutic target signatures

Studies of a Vascular-Acting Photosensitizer, Pd-Bacteriopheophorbide (Tookad), in Normal Canine Prostate and Spontaneous Canine Prostate Cancer

Quantum Dots for Multiplexed Detection and Characterisation of Prostate Cancer Cells Using a Scanning Near-Field Optical Microscope

Histotripsy Focal Ablation of Implanted Prostate Tumor in an ACE-1 Canine Cancer Model

In vitro and in vivo model systems used in prostate cancer research

CCR4 blockade leads to clinical activity and prolongs survival in a canine model of advanced prostate cancer

Urethral Sparing Histotripsy of the Prostate in a Canine Model

Generation and Characterisation of a Canine EGFP-HMGA2 Prostate Cancer In Vitro Model

Similar Documents

CircTHSD4 promotes the malignancy and docetaxel (DTX) resistance in prostate cancer by regulating miR-203/HMGA2 axis

Generation of a syngeneic orthotopic transplant model of prostate cancer metastasis

RNA-seq of nine canine prostate cancer cell lines reveals diverse therapeutic target signatures

Studies of a Vascular-Acting Photosensitizer, Pd-Bacteriopheophorbide (Tookad), in Normal Canine Prostate and Spontaneous Canine Prostate Cancer

Quantum Dots for Multiplexed Detection and Characterisation of Prostate Cancer Cells Using a Scanning Near-Field Optical Microscope

Histotripsy Focal Ablation of Implanted Prostate Tumor in an ACE-1 Canine Cancer Model

In vitro and in vivo model systems used in prostate cancer research

CCR4 blockade leads to clinical activity and prolongs survival in a canine model of advanced prostate cancer

Urethral Sparing Histotripsy of the Prostate in a Canine Model

Generation and Characterisation of a Canine EGFP-HMGA2 Prostate Cancer In Vitro Model