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Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS

10 ARTICLES PUBLISHED IN JoVE

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Cancer Research

Cell-Free DNA Integrity Analysis in Urine Samples
Valentina Casadio 1, Samanta Salvi 1, Filippo Martignano 1, Roberta Gunelli 2, Sara Ravaioli 1, Daniele Calistri 1
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Urology, Morgagni-Pierantoni Hospital

A method for analyzing DNA integrity in the cell-free supernatant fraction of urine samples is proposed. The method is suitable for early detection of urological malignancies and has proven accurate for the early diagnosis of bladder cancer.

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Cancer Research

Development of a Human Preclinical Model of Osteoclastogenesis from Peripheral Blood Monocytes Co-cultured with Breast Cancer Cell Lines
Laura Mercatali 1, Chiara Spadazzi 1, Giacomo Miserocchi 1, Chiara Liverani 1, Alessandro De Vita 1, Alberto Bongiovanni 1, Federica Recine 1, Dino Amadori 1, Toni Ibrahim 1
1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), IRCCS

This protocol describes development of an in vitro human preclinical model of osteoclastogenesis from peripheral blood monocytes cultured with breast cancer cell lines to mimic the cancer cell-osteoclast interaction. The model could be used to further our understanding of bone metastasis formation and improve therapeutic options.

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Cancer Research

Serum and Plasma Copy Number Detection Using Real-time PCR
Samanta Salvi 1, Vincenza Conteduca 1, Filippo Martignano 1, Giorgia Gurioli 1, Daniele Calistri 1, Valentina Casadio 1
1Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS

This manuscript describes the copy number variation analysis performed in serum or plasma DNA using real-time PCR approach. This method is suitable for the prediction of drug resistance in castration resistant prostate cancer patients, but it could be informative also for other diseases.

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Cancer Research

Looking for Driver Pathways of Acquired Resistance to Targeted Therapy: Drug Resistant Subclone Generation and Sensitivity Restoring by Gene Knock-down
Chiara Arienti 1, Sara Pignatta 1, Michele Zanoni 1, Michela Cortesi 1, Alice Zamagni 1, Filippo Piccinini 2, Anna Tesei 1
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS

This is a time- and cost-effective in vitro protocol investigating the mechanisms of acquired resistance to targeted therapeutic agents, which is a highly unmet medical need in cancer management.

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Cancer Research

Establishment of a Primary Culture of Patient-derived Soft Tissue Sarcoma
Alessandro De Vita 1, Laura Mercatali 1, Giacomo Miserocchi 1, Chiara Liverani 1, Chiara Spadazzi 1, Federica Recine 1, Alberto Bongiovanni 1, Federica Pieri 2, Davide Cavaliere 3, Valentina Fausti 1, Dino Amadori 1, Toni Ibrahim 1
1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Pathology Unit, Morgagni-Pierantoni Hospital, 3Unit of Surgery and Advanced Oncologic Therapies, Morgagni-Pierantoni Hospital

The following protocol focuses on the establishment of a primary culture of patient-derived soft tissue sarcoma (STS). This model could help us to better understand the molecular background and pharmacological profile of these rare malignancies and could represent a starting point for further research aimed at improving STS management.

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Cancer Research

Characterization of Tumor Cells Using a Medical Wire for Capturing Circulating Tumor Cells: A 3D Approach Based on Immunofluorescence and DNA FISH
Giulia Gallerani 1, Claudia Cocchi 2, Martine Bocchini 3, Filippo Piccinini 1, Francesco Fabbri 1
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Associazione Annastaccatolisa Onlus, 3Nuclear Medicine Unit, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS

We present a new approach to characterize tumor cells. We combined immunofluorescence with DNA fluorescent-in-situ-hybridization to evaluate cells captured by a functionalized medical wire capable of in vivo enriching CTCs directly from patient blood.

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Cancer Research

Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR
Chiara Molinari 1, Raefa Abou Khouzam 2, Samanta Salvi 1, Tania Rossi 1, Guglielmina Nadia Ranzani 2, Daniele Calistri 1
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Biology and Biotechnology, University of Pavia

The manuscript describes a chip-based digital PCR assay to detect a rare CDH1 transcript variant (CDH1a) in fresh-frozen normal and tumor tissues obtained from patients with gastric cancer.

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Cancer Research

Dried Blood and Serum Spots As A Useful Tool for Sample Storage to Evaluate Cancer Biomarkers
Laura Mercatali 1, Patrizia Serra 2, Giacomo Miserocchi 1, Chiara Spadazzi 1, Chiara Liverani 1, Alessandro De Vita 1, Giorgia Marisi 3, Alberto Bongiovanni 1, Federica Recine 1, Akwilina Pangan 4, Nestory Masalu 4, Toni Ibrahim 1, Dino Amadori 1
1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 2Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 3Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 4Department of Oncology, Bugando Medical Center, Mwanza, Tanzania

This protocol describes a simple and useful method to store peripheral blood and serum/plasma for downstream analyses such as single nucleotide polymorphism (SNP) evaluation and ELISA assay.

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Cancer Research

Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer
Matteo Canale 1, Giorgia Marisi 1, Alessandro Passardi 2, Emanuela Scarpi 3, Paola Ulivi 1
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Medical Oncology, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 3Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS

We present a protocol to evaluate the expression levels of circulating microRNA (miRNAs) in plasma samples from cancer patients. In particular, we used a commercially available kit for circulating miRNA extraction and reverse transcription. Finally, we analyzed a panel of 24 selected miRNAs using real-time pre-spotted probe custom plates.

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Cancer Research

Evaluation of Colorectal Cancer Risk and Prevalence by Stool DNA Integrity Detection
Claudia Rengucci 1, Giulia De Maio 1, Maura Menghi 2, Francesca Benzi 2, Daniele Calistri 1
1Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 2Diatech Pharmacogenetics srl

The presented diagnostic FL-DNA kit is a time-saving and user-friendly method to determine the reliable probability of the presence of colorectal cancer lesions.

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