Gene expression analysis of bone metastasis and circulating tumor cells from metastatic castrate-resistant prostate cancer patients
| Autor: | Won Jin Cho, Michael L. Cher, Hyeong Reh Choi Kim, Leandro E. Mainetti, Elisabeth I. Heath, R. Daniel Bonfil, Abdo J. Najy, Hussein D. Aoun, Daniel S. M. Oliveira |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male Pathology LASER CAPTURE MICRODISSECTION Biopsy PROSTATE CANCER Prostate cancer 0302 clinical medicine Circulating tumor cell Bone Marrow Gene expression Medicine RNA Neoplasm Medicine(all) Aged 80 and over Bone metastasis General Medicine Laser capture microdissection purl.org/becyt/ford/3.1 [https] Middle Aged Neoplastic Cells Circulating BONE METASTASIS Antisense RNA Gene Expression Regulation Neoplastic Medicina Básica Prostatic Neoplasms Castration-Resistant medicine.anatomical_structure 030220 oncology & carcinogenesis purl.org/becyt/ford/3 [https] medicine.medical_specialty GENE EXPRESSION CIENCIAS MÉDICAS Y DE LA SALUD Inmunología Bone Neoplasms General Biochemistry Genetics and Molecular Biology CIRCULATING TUMOR CELLS 03 medical and health sciences Cell Line Tumor Humans RNA Antisense RNA Messenger Gene Aged Biochemistry Genetics and Molecular Biology(all) business.industry Research Gene Expression Profiling Circulating tumor cells Reproducibility of Results medicine.disease Gene expression profiling 030104 developmental biology Bone marrow business |
| Zdroj: | CONICET Digital (CONICET) Consejo Nacional de Investigaciones Científicas y Técnicas instacron:CONICET Journal of Translational Medicine |
| DOI: | 10.1186/s12967-016-0829-5 |
| Popis: | Background: Characterization of genes linked to bone metastasis is critical for identification of novel prognostic or predictive biomarkers and potential therapeutic targets in metastatic castrate-resistant prostate cancer (mCRPC). Although bone marrow core biopsies (BMBx) can be obtained for gene profiling, the procedure itself is invasive and uncommon practice in mCRPC patients. Conversely, circulating tumor cells (CTCs), which are likely to stem from bone metastases, can be isolated from blood. The goals of this exploratory study were to establish a sensitive methodology to analyze gene expression in BMBx and CTCs, and to determine whether the presence or absence of detectable gene expression is concordant in matching samples from mCRPC patients. Methods: The CellSearch® platform was used to enrich and enumerate CTCs. Low numbers of PC3 prostate cancer (PCa) cells were spiked into normal blood to assess cell recovery rate. RNA extracted from recovered PC3 cells was amplified using an Eberwine-based procedure to obtain antisense mRNA (aRNA), and assess the linearity of the RNA amplification method. In this pilot study, RNAs extracted from CTCs and PCa cells microdissected from formalin-fixed paraffin-embedded BMBx, were amplified to obtain aRNA and assess the expression of eight genes functionally relevant to PCa bone metastasis using RT-PCR. Results: RNAs were successfully extracted from as few as 1-5 PCa cells in blood samples. The relative expression levels of reference genes were maintained after RNA amplification. The integrity of the amplified RNA was also demonstrated by RT-PCR analysis using primer sets that target the 5'-end, middle, and 3'-end of reference mRNA. We found that in 21 out of 28 comparisons, the presence or absence of detectable gene expression in CTCs and PCa cells microdissected from single bone lesions of the same patients was concordant. Conclusions: This exploratory analysis suggests that aRNA amplification through in vitro transcription may be useful as a method to detect gene expression in small numbers of CTCs and tumor cells microdissected from bone metastatic lesions. In some cases, gene expression in CTCs and BMBxs was not concordant, raising questions about using CTC gene expression to make clinical decisions. Fil: Cho, Won Jin. Wayne State University; Estados Unidos Fil: Oliveira, Daniel S.M.. Wayne State University; Estados Unidos Fil: Najy, Abdo J.. Wayne State University; Estados Unidos Fil: Mainetti, Leandro Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Wayne State University; Estados Unidos Fil: Aoun, Hussein D.. Wayne State University; Estados Unidos Fil: Cher, Michael L.. Wayne State University; Estados Unidos Fil: Heath, Elisabeth. Wayne State University; Estados Unidos Fil: Kim, Hyeong-Reh C.. Wayne State University; Estados Unidos Fil: Bonfil, Ricardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Wayne State University; Estados Unidos |
| Databáze: | OpenAIRE |
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