Lack of expression of preproorexin and orexin receptors genes in human normal and prostate cancer cell lines

Abstract

Introduction. Studies on expression of orexins (OXs) and their receptors in human prostate gland and human prostatic cell lines are scanty and their results contradictory. Regarding this, we carefully reinvestigated this problem on human prostatic cell lines. Material and methods. Expression of preproorexin (ppOX) (6 primer pairs), and orexin receptors 1 and 2 (OXR1, OXR2) (4 and 2 primer pairs, respectively) was assessed by conventional PCR and QPCR in human normal (PrEC, PrSc, PrSmC) and prostate carcinoma (Du145, LNCaP, and PC3) cell lines. We designed intron spanning primers and also we applied primers from earlier publications and commercially available ones. Results. With the designed primer pairs, in all studied cell lines we failed to demonstrate expression of ppOX, OXR1 and OXR2 genes at the mRNA level, while reaction products were observed in control tissues (human placenta and adrenals). Primers applied in earlier studies did not form amplification products specific for preproorexin or orexin 1 receptor. Some commercially available primers for orexin receptor 1 produced false positive results. Conclusions. We found no evidence for the presence of preproorexin-orexin receptors system genes' mRNAs in human prostate cell lines. The reported premises for these genes' expression in prostate and prostatic cell lines may have arisen either from the presence of non-prostate cells included in the samples or from faulty PCR settings.

ZFP91: A Noncanonical NF-κB Signaling Pathway Regulator with Oncogenic Properties Is Overexpressed in Prostate Cancer

“…Analyses were performed as described earlier [2, 18, 19]. Briefly, primers were designed by Primer 3 software (Whitehead Institute for Biomedical Research, Cambridge, USA) and purchased from the Laboratory of DNA Sequencing and Oligonucleotide Synthesis (Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland).…”

A review of orexin’s unprecedented potential as a novel, highly-specific treatment for various localized and metastatic cancers

“…OX1R mRNA was expressed in all clinical prostate cancer samples tested. 15 The expression of OX1R mRNA was variable in LNCaP 9 , 15 , 16 and DU145 9, 16 cell lines. OX1R mRNA was not found in the following cell lines: PC3, 16 PrEC, 16 PrSmC, 16 PrSc, 16 or in any clinical normal and benign prostatic hyperplasia (BPH) sample tissues 15 tested.…”

“… 15 The expression of OX1R mRNA was variable in LNCaP 9 , 15 , 16 and DU145 9, 16 cell lines. OX1R mRNA was not found in the following cell lines: PC3, 16 PrEC, 16 PrSmC, 16 PrSc, 16 or in any clinical normal and benign prostatic hyperplasia (BPH) sample tissues 15 tested. OX2R mRNA was expressed in all clinical and BPH sample tissues, 15 but was not expressed in LNCaP, 9 , 16 DU145, 9 , 16 PC3, 16 PrEC, 16 PrSmC, 16 or PrSc 16 cell lines.…”

ZFP91 zinc finger protein expression pattern in normal tissues and cancers

“…Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis. Analyses were performed as described earlier (14,(17)(18)(19). Briefly, primers were designed by Primer 3 software (Whitehead Institute for Biomedical Research, Cambridge, USA) and purchased from the Laboratory of DNA Sequencing and Oligonucleotide Synthesis (Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland).…”