Description(s):

miRNAs have been implicated in the regulation of various biological processes such as cell death and cell proliferation. They can act as oncogenes and tumor suppressor genes, playing a crucial role in tumorogenesis. The change in the expression level of miRNAs is associated with dysfunction of their corresponding protein-coding gene targets, many of which are involved in the initiation and progression of cancer. For example, BCL2 oncogene is targeted by miR-15a and miR-16, while PTEN tumor suppressor is targeted by miR-21, and HOXD10, Pak, MYOD1 and ER, and Her2 are targeted by miR-10b, miR-7, miR-206, and miR-125a, respectively.
Signosis developed cancer miRNA plate assay kit I for quantitatively profiling the expression of seven well-known cancer related miRNAs, which are miRNA-15a, miR-16, miR-21, miR10b, miR-7, miR-125a and miR206. RNU48 is included for normalization.

Principle

In the proprietary miRNA plate assay, one miRNA molecule is flanked by a capture oligo and a biotinated detection oligo through two bridges oligos. One of the bridge oligos is partially hybridized with the miRNA molecule and the capture oligo and another with the miRNA and the detection oligo. The hybrid is immobilized onto plate through hybridization with an immobilized oligo and detected by a streptavidin-HRP conjugate and chemiluminecscent substrate. This hybrid structure is sensitive to the sequence of the miRNA molecule. One nucleotide difference will prevent the formation of the hybrid and therefore miRNA isoform can be differentiated, which normally is hard to tackle with Northern blot. In addition, the sensitivity of the assay is higher than miRNA Northern blot assay.

Data

Expression of miRNAs was analyzed with 0.5ug total RNA from normal liver andT47D cells respectively. The assay was subjected to chemiluminescent plate reader.

Literature

View user manual

Citations

HMGA2 Overexpression-Induced Ovarian Surface Epithelial Transformation Is Mediated Through Regulation of EMT Genes. Wu J, Liu Z, Shao C, Gong Y, Hernando E, Lee P, Narita M, Muller W, Liu J, Wei JJ.Cancer Res. 2011 Jan 15;71(2):349-59.