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Cancer Research

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Nanyang Technological University

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

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13.6K Views

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09:58 min

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December 9th, 2016

DOI :

10.3791/54714-v

December 9th, 2016

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Rocco Sciarrillo¹^,²^,³, Anna Wojtuszkiewicz¹, Irsan E. Kooi⁴, Valentina E. Gómez³, Ugo Boggi⁵, Gerrit Jansen⁶, Gert-Jan Kaspers¹^,⁷, Jacqueline Cloos*¹, Elisa Giovannetti*³^,⁸^,⁹

¹Department of Pediatric Oncology/Hematology, VU University Medical Center, ²Department of Hematology, VU University Medical Center, ³Department of Medical Oncology, VU University Medical Center, ⁴Department of Clinical Genetics, VU University Medical Center, ⁵Division of General and Transplant Surgery, Azienda Ospedaliera Universitaria Pisana, Universita’ di Pisa, ⁶Amsterdam Immunology and Rheumatology Center, VU University Medical Center, ⁷Princess Máxima Center for Pediatric Oncology, ⁸Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, ⁹Institute of Nanoscience and Nanotechnology, CNR-Nano

Podsumowanie

Here we describe a protocol aimed at investigating the impact of aberrant splicing on drug resistance in solid tumors and hematological malignancies. To this goal, we analyzed the transcriptomic profiles of parental and resistant in vitro models through RNA-seq and established a qRT-PCR based method to validate candidate genes.

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Splice Variants

Drug Resistance

In Vitro Cancer Models

Molecular Pharmacology

Chemotherapy Resistance

Transcriptomic Analysis

Cytotoxicity Screening

Formazan Crystals

Microplate Reader

Rozdziały w tym wideo

0:05

Title

1:09

Characterization of Drug Resistance Profiles through Cytotoxicity Assays

5:01

RNA Isolation and Library Preparation for RNA-sequencing

5:55

Validation of the Results by RT-PCR and qRT-PCR Assays

7:21

Results: Detection of Differential Splicing Events in Chemotherapy-resistant Cell Lines Compared to Parental Wild-type by Using RNA-sequencing

9:07

Conclusion

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