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105 Identification and characterization of long noncoding RNA markers for prostate cancer detection
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Identification and characterization of long noncoding RNA markers for prostate cancer detection Eur Urol Suppl 2014;13;e105
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Lee B. 1 , Mazar J. 1 , Aftab M. 1 , Qi F.1 , Li J-L. 1 , Govindarajan S. 1 , Kameh D.2 , Patel V. 3 , Perera R.1 1 Sanford-Burnham 2 Florida
Medical Research Institute, Dept. of Analytical Genomics and Bioinformatics, Orlando, United States of America,
Hospital, Celebration Health, Dept. of Pathology, Celebration, United States of America, 3 Florida Hospital, Global Robotics Institute,
Dept. of Urology, Celebration, United States of America INTRODUCTION & OBJECTIVES: Prostate cancer (PCa) is one of the leading causes of cancer deaths among American men. Digital rectal exam and prostate-specific antigen (PSA) are the most commonly used screening methods for PCa. However, several advisory groups now recommend against use of the PSA tests because the benefits are, at best, small. Thus, there is an urgent unmet need for novel and accurate diagnostic biomarkers for PCa to reduce overtreatment and associated morbidity. We therefore conducted a study to identify differentially expressed long noncoding RNAs (lncRNAs) in PCa cell lines and patient samples using DNA microarrays, and performed confirmatory analysis using qRT-PCR and RNA-FISH. MATERIAL & METHODS: Several highly upregulated lncRNAs were further tested in patient prostatic adenocarcinoma tissue samples (Gleason score >6.0) and compared to matched normal tissues. AK024556, XLOC-007697, LOC100506411, LOC100287482, XLOC001699, XLOC-005327, XLOC-008559, and XLOC-009911 were confirmed as highly upregulated in patient samples. RESULTS: AK024556, also known as SPRY4-IT1, an intronic lncRNA originating from the first intron of the SPRY4 gene was previously reported to be upregulated in primary human melanomas and melanoma cell lines. SPRY4-IT1 was not expressed in LNCaP cells due to the epigenetic modification (CpG island methylation) of the SPRY4 promoter. Epigenetic silencing was reversed by treatment with 5-aza-2’deoxycytidine (a DNA methyltransferase inhibitor) and resulted in upregulation of SPRY4 and SPRY4-IT1, indicating that SPRY4 and SPRY4-IT1 are epigenetically co-regulated. siRNA knockdown of SPRY4-IT1 inhibited proliferation and invasion, and increased apoptosis in PC3 cells. Chromogenic in situ hybridization (CISH) assay was developed to detect SPRY4-IT1 in PCa, and we believe that this assay will be useful for prostate cancer diagnosis in a clinical setting. CONCLUSIONS: We report here support that lncRNAs are potential molecular markers for prostate cancer detection in men.
Identification and characterization of long noncoding RNA markers for prostate cancer detection Eur Urol Suppl 2014;13;e105
Print! Print!
Lee B. 1 , Mazar J. 1 , Aftab M. 1 , Qi F.1 , Li J-L. 1 , Govindarajan S. 1 , Kameh D.2 , Patel V. 3 , Perera R.1 1 Sanford-Burnham 2 Florida
Medical Research Institute, Dept. of Analytical Genomics and Bioinformatics, Orlando, United States of America,
Hospital, Celebration Health, Dept. of Pathology, Celebration, United States of America, 3 Florida Hospital, Global Robotics Institute,
Dept. of Urology, Celebration, United States of America INTRODUCTION & OBJECTIVES: Prostate cancer (PCa) is one of the leading causes of cancer deaths among American men. Digital rectal exam and prostate-specific antigen (PSA) are the most commonly used screening methods for PCa. However, several advisory groups now recommend against use of the PSA tests because the benefits are, at best, small. Thus, there is an urgent unmet need for novel and accurate diagnostic biomarkers for PCa to reduce overtreatment and associated morbidity. We therefore conducted a study to identify differentially expressed long noncoding RNAs (lncRNAs) in PCa cell lines and patient samples using DNA microarrays, and performed confirmatory analysis using qRT-PCR and RNA-FISH. MATERIAL & METHODS: Several highly upregulated lncRNAs were further tested in patient prostatic adenocarcinoma tissue samples (Gleason score >6.0) and compared to matched normal tissues. AK024556, XLOC-007697, LOC100506411, LOC100287482, XLOC001699, XLOC-005327, XLOC-008559, and XLOC-009911 were confirmed as highly upregulated in patient samples. RESULTS: AK024556, also known as SPRY4-IT1, an intronic lncRNA originating from the first intron of the SPRY4 gene was previously reported to be upregulated in primary human melanomas and melanoma cell lines. SPRY4-IT1 was not expressed in LNCaP cells due to the epigenetic modification (CpG island methylation) of the SPRY4 promoter. Epigenetic silencing was reversed by treatment with 5-aza-2’deoxycytidine (a DNA methyltransferase inhibitor) and resulted in upregulation of SPRY4 and SPRY4-IT1, indicating that SPRY4 and SPRY4-IT1 are epigenetically co-regulated. siRNA knockdown of SPRY4-IT1 inhibited proliferation and invasion, and increased apoptosis in PC3 cells. Chromogenic in situ hybridization (CISH) assay was developed to detect SPRY4-IT1 in PCa, and we believe that this assay will be useful for prostate cancer diagnosis in a clinical setting. CONCLUSIONS: We report here support that lncRNAs are potential molecular markers for prostate cancer detection in men.