Expression Analysis of BORIS during Retinoic Acid Induced Differentiation of Human Embryonal Carcinoma NCCIT Cell Line

Background: BORIS belongs a group of genes that are expressed normally in testis, and abnormally in various types of cancers. Recently, expression of BORIS in cancer stemcells and its role in expression induction of some CSC markers and in the regulation of CSC features has been demonstrated. Embryonic carcinoma (EC) cell line are thepluripotent cells derived from tumors known as teratocarcinomas and are generally considered to be the archetype of cancer stem cells. Materials/Patients and Methods: NCCIT cell line were cultured in RPMI supplemented with 10% fetal bovine serum. To induce differentiation, NCCIT cells were treated with 10 μM RA for 21 d. Primers for detection of all isoform of BORIS as well as the β2M gene as a reference were designed by oligo 7 software. After cDNA synthesis, one microliters of complementary DNA products were amplified with SYBR Green real-time PCR Master Mix. Quantification of target genes was normalized using the β2M gene as a reference and subsequently expressed as relative to untreated control cells. Fold change was calculated according to the following equation: 2(-ΔΔCT).Results: Although designed primer for BORIS can amplify all isoform for BORIS, Real time PCR did not show expression of BORIS in NCCIT cell line. Expression of BORISwas not induced during RA-induced differentiation of NCCIT cell line. Conclusion: In fact, NCCIT cell line is a heterogeneous population of cells comprising a small population of BORIS-expressing cancer stem cells. Therefore, in order to investigate expression of BORIS in this cell line, at first stem-like cells must be first isolated according to their markers and properties. There for it can be probable that heterogeneous population of NCCIT do not show expression of this gene.

کلیدواژه ها:

BORIS ، Human Embryonal Carcinoma NCCIT Cell Line ، Retinoic acid ، Differentiation