Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples

Abstract

The Saint James's Hospital Biobank was established in 2008, to develop a high-quality breast tissue BioResource, as a part of the breast cancer clinical care pathway. The aims of this work were: (1) to ascertain the quality of RNA, DNA, and protein in biobanked carcinomas and normal breast tissues, (2) to assess the efficacy of AllPrep(®) (Qiagen) in isolating RNA, DNA, and protein simultaneously, (3) to compare AllPrep with RNEasy(®) and QIAamp(®) (both Qiagen), and (4) to examine the effectiveness of Allprotect(®) (Qiagen), a new tissue stabilization medium in preserving DNA, RNA, and proteins. One hundred eleven frozen samples of carcinoma and normal breast tissue were analyzed. Tumor and normal tissue morphology were confirmed by frozen sections. Tissue type, tissue treatment (Allprotect vs. no Allprotect), extraction kit, and nucleic acid quantification were analyzed by utilizing a 4 factorial design (SPSS PASW 18 Statistics Software(®)). QIAamp (DNA isolation), AllPrep (DNA, RNA, and Protein isolation), and RNeasy (RNA isolation) kits were assessed and compared. Mean DNA yield and A(260/280) values using QIAamp were 33.2 ng/μL and 1.86, respectively, and using AllPrep were 23.2 ng/μL and 1.94. Mean RNA yield and RNA Integrity Number (RIN) values with RNeasy were 73.4 ng/μL and 8.16, respectively, and with AllPrep were 74.8 ng/μL and 7.92. Allprotect-treated tissues produced higher RIN values of borderline significance (P=0.055). No discernible loss of RNA stability was detected after 6 h incubation of stabilized or nonstabilized tissues at room temperature or 4°C or in 9 freeze-thaw cycles. Allprotect requires further detailed evaluation, but we consider AllPrep to be an excellent option for the simultaneous extraction of RNA, DNA, and protein from tumor and normal breast tissues. The essential presampling procedures that maintain the diagnostic integrity of pathology specimens do not appear to compromise the quality of molecular isolates.

Development and Progress of Ireland's Biobank Network: Ethical, Legal, and Social Implications (ELSI), Standardized Documentation, Sample and Data Release, and International Perspective

“…Our group previously described quality control in breast cancer biobanking. 10 The SJH biobank provides training and guidance to hospitals entering the network.…”

“…12 Although the SAP was only finalized in 2011, three projects have been approved, and have already led to scientific publications. 10,53 Data Protection and the Irish National Cancer Registry…”

Can a biobank network and supporting infrastructure enhance Ireland's ability to attract pharmaceutical research and development and clinical trial opportunities? A pilot survey

“…The network, which commenced in 2008, aims to deliver benefits for patients, researchers, industry, and the economy. 13,14 BIT engages with stakeholders, including patient advocate groups, who are positive about industry accessing their donated biobanked material (blood, tissue, etc), because of the hope of new and improved treatments. BIT's network, which is under development, could also act as an interface between industry and academia, to assist in fostering collaborations.…”

Utilização do produto Allprotect Tissue Reagent® na estabilização do DNA extraído de tecidos dentais humanos em diferentes condições de armazenamento

“…Neste estudo foi possível observar que nos dentes íntegros mantidos refrigerados não houve uma melhoria significativa na quantidade de DNA ao se utilizar o produto, assim como no armazenamento da polpa dental, em que o uso ou não do produto não evidenciou diferenças significativas em ambas às condições de temperatura. Esses resultados são similares ao encontrado no estudo de Mee et al (2011), que não observaram diferenças na qualidade do DNA extraído ao comparar a utilização ou não do produto, antes do congelamento em amostra de tecidos mamários.…”

“…Assim, torna-se possível que as evidências recolhidas em qualquer lugar à temperatura ambiente sejam enviadas para as instalações apropriadas de forma segura (Grotzer et al, 2000;Medeiros et al, 2003;Mutter et al, 2004;Qiagen, 2006Qiagen, , 2008Qiagen, , 2011 (Mee et al, 2011;Gelbart et al, 2012), ainda não há trabalhos na literatura que evidenciam a funcionalidade e eficácia do mesmo em relação aos tecidos dentais.…”