Author/Authors :
Poursani, Ensieh M Department of Molecular Genetics - Faculty of Biological Sciences - Tarbiat Modares University, Tehran, Iran , Mehravar, Majid Department of Molecular Genetics - Faculty of Biological Sciences - Tarbiat Modares University, Tehran, Iran , Mohammad Soltani, Bahram Department of Molecular Genetics - Faculty of Biological Sciences - Tarbiat Modares University, Tehran, Iran , Mowla, Javad Department of Molecular Genetics - Faculty of Biological Sciences - Tarbiat Modares University, Tehran, Iran , Trosko, James E Food Safety Toxicology Center - Department of Pediatrics and Human Development - Michigan State University, East Lansing, Michigan (MI), USA
Abstract :
Background: Alternative splicing is an important mechanism that regulates gene expression
and function in human cells. OCT4, a crucial pluripotency marker in embryonic
stem/carcinoma cells generates several spliced variants in different cell types and
cancers. The expression of OCT4 in cancers has been challenged in many studies. The
existence of several OCT4 spliced variants and absence of specific discriminating primers
is the main reason of this controversy. Therefore, using specific primers and discriminating
OCT4 variants from each other might help to reduce these discrepancies
in carcinogenesis and stem cell researches.
Methods: 17 various human cancer, pluripotent and normal cells were cultured and
their RNAs were extracted. Related cDNAs were synthesized and the expression pattern
of OCT4 variants was investigated by RT-PCR assay. PCR products were cloned
into pTZ57R/T vector and their authenticity was confirmed by DNA sequencing.
Results: Expression pattern of OCT4 variants (OCT4A, OCT4B and OCT4B1) was analyzed
by RT-PCR assay and the authenticity of PCR products was confirmed by DNA
sequencing. A novel spliced variant of OCT4 was discovered and named as OCT4B3.
This variant was very similar to OCT4B2 transcript except that 207-nt of exon 1b is
lost. Moreover, the expression pattern of OCT4B3 variant was investigated in 17 human
cell types, where its expression was only found in astrocytoma and bladder cancer
cell types 1321N1 and 5637, respectively.
Conclusion: OCT4 variants are differentially expressed in various human cancer cell
lines. Moreover, a novel variant of OCT4, OCT4B3, was detected in two human cancer
cell lines of bladder carcinoma (5637) and brain astrocytoma (1321N1) for the first
time.
Keywords :
Stem cells , Gene , Cell lines , Alternative splicing