Poursani Ensieh M, Mehravar Majid, Mohammad Soltani Bahram, Mowla Seyed Javad, Trosko James E
Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Food Safety Toxicology Center, Department of Pediatrics and Human Development, Michigan State University, East Lansing, Michigan (MI), USA.
Avicenna J Med Biotechnol. 2017 Jul-Sep;9(3):142-145.
Alternative splicing is an important mechanism that regulates gene expression and function in human cells. , a crucial pluripotency marker in embryonic stem/carcinoma cells generates several spliced variants in different cell types and cancers. The expression of in cancers has been challenged in many studies. The existence of several spliced variants and absence of specific discriminating primers is the main reason of this controversy. Therefore, using specific primers and discriminating variants from each other might help to reduce these discrepancies in carcinogenesis and stem cell researches.
17 various human cancer, pluripotent and normal cells were cultured and their RNAs were extracted. Related cDNAs were synthesized and the expression pattern of variants was investigated by RT-PCR assay. PCR products were cloned into pTZ57R/T vector and their authenticity was confirmed by DNA sequencing.
Expression pattern of 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 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.
variants are differentially expressed in various human cancer cell lines. Moreover, a novel variant of , OCT4B3, was detected in two human cancer cell lines of bladder carcinoma (5637) and brain astrocytoma (1321N1) for the first time.
可变剪接是调节人类细胞中基因表达和功能的重要机制。OCT4是胚胎干/癌细胞中的关键多能性标志物，在不同细胞类型和癌症中产生多种剪接变体。许多研究对OCT4在癌症中的表达提出了质疑。存在多种OCT4剪接变体且缺乏特异性区分引物是造成这一争议的主要原因。因此，使用特异性引物并区分彼此的OCT4变体可能有助于减少癌症发生和干细胞研究中的这些差异。
培养17种不同的人类癌症、多能性和正常细胞，并提取其RNA。合成相关cDNA，通过RT-PCR分析研究OCT4变体的表达模式。将PCR产物克隆到pTZ57R/T载体中，并通过DNA测序确认其真实性。
通过RT-PCR分析OCT4变体（OCT4A、OCT4B和OCT4B1）的表达模式，并通过DNA测序确认PCR产物的真实性。发现了一种新的OCT4剪接变体，命名为OCT4B3。该变体与OCT4B2转录本非常相似，只是外显子1b的207个核苷酸缺失。此外,在17种人类细胞类型中研究了OCT4B3变体的表达模式，结果发现其仅分别在星形细胞瘤以及膀胱癌细胞系1321N1和5637中表达。
OCT4变体在各种人类癌细胞系中差异表达。此外，首次在两种人类膀胱癌细胞系（5637）和脑星形细胞瘤（1321N1）中检测到一种新的OCT4变体OCT4B3。
