Article Text
Abstract
Aims: To elucidate genes that participate in the process of oncogenesis, primers based on the E6 genes of genital human papillomaviruses (HPVs) were used to amplify potential expressed sequence tags (ESTs) from the MOLT-4 T lymphoblastic leukaemia cell line.
Methods: Using the polymerase chain reaction (PCR) with human papillomavirus E6 gene primers, an EST from the MOLT-4 T lymphoblastic leukaemia cell line was amplified. Via rapid amplification of cDNA ends (RACE) and cycle sequencing from MOLT-4 and fetal lung cDNA libraries, overlapping cDNAs of 2786 bp and 2054 bp of the corresponding novel human intronless gene designated MOST-1 (for MOLT-4 sequence tag-1) were characterised and assigned the symbol C8orf17 by the HUGO Nomenclature Committee.
Results: Both cDNAs contained a potential open reading frame (ORF) of 297 bp incorporating a methionine codon with an ideal Kozak consensus sequence for translation initiation, and encoding a putative hydrophilic polypeptide of 99 amino acids. Although reverse transcription PCR (RT-PCR) demonstrated MOST-1 expression in all 19 cancer and two normal cell lines tested, differential expression was seen in only nine of 16 normal tissues tested (heart, kidney, liver, pancreas, small intestine, ovary, testis, prostate, and thymus). A 388 bp fragment was amplified from the NS-1 mouse myeloma cell line, the sequence of which was identical to that within the MOST-1 ORF. The MOST-1 gene was mapped by fluorescent in situ hybridisation to chromosome 8q24.2, a region amplified in many breast cancers and prostate cancers, which is also the candidate site of potential oncogene(s) other than c-myc located at 8q24.1. Analysis of paired biopsies of invasive ductal breast cancer and adjacent normal tissue by semiquantitative and real time RT-PCR revealed average tumour to normal ratios of MOST-1 expression that were two times greater in grade 3 cancers than in grade 1 and 2 cancers. Quantitative real time PCR of archival prostatic biopsies displayed MOST-1 DNA values that were 9.9, 7.5, 4.2, and 1.4 times higher in high grade carcinomas, intermediate grade carcinomas, low grade carcinomas, and benign hyperplasias, respectively, than in normal samples.
Conclusions: These data suggest a role for MOST-1 in cellular differentiation, proliferation, and carcinogenesis.
- amplification
- breast cancer
- C8orf17
- MOST-1
- overexpression
- prostate cancer
- CT, threshold cycle
- EST, expressed sequence tag
- FISH, fluorescence in situ hybridisation
- G3PDH, glyceraldehyde-3-phosphate dehydrogenase
- HPV, human papillomavirus
- nt, nucleotides
- N, normal
- ORF, open reading frame
- PBS, phosphate buffered saline
- PCR, polymerase chain reaction
- RACE, rapid amplification of cDNA ends
- RT, reverse transcription
- SDS, sodium dodecyl sulfate
- T, tumour
- UTR, untranslated region
Statistics from Altmetric.com
- CT, threshold cycle
- EST, expressed sequence tag
- FISH, fluorescence in situ hybridisation
- G3PDH, glyceraldehyde-3-phosphate dehydrogenase
- HPV, human papillomavirus
- nt, nucleotides
- N, normal
- ORF, open reading frame
- PBS, phosphate buffered saline
- PCR, polymerase chain reaction
- RACE, rapid amplification of cDNA ends
- RT, reverse transcription
- SDS, sodium dodecyl sulfate
- T, tumour
- UTR, untranslated region