Characterization

Description(s):

MiRNAs have been widely known to function as either oncogenes or tumor suppressors under certain conditions. The dysregulated miRNAs have been shown to affect the hallmarks of cancer, including sustaining proliferative signaling, eluding growth suppressors, resisting cell death, and inducing angiogenesis. An increasing number of studies have identified miRNAs as potential biomarkers for human cancer diagnosis, prognosis and therapeutic targets. Signosis has developed Cancer miRNA Array including 132 cancer-related miRNAs for cancer research.

Applicable Grid:

List of Applicable miRNAs

Let-7a-5p	Let-7b-5p	Let-7c-5p	Let-7d-5p	Let-7e-5p	Let-7f-5p	Let-7g-5p	Let-7i-5p	miR-1	miR- 7
miR-9-5p	miR-10a-5p	miR-15a-5p	miR-15b-5p	miR-16-5p	miR-17-5p	miR-18b-5p	miR- 18b	miR-19a-3p	miR-19b-3p
miR-20a-5p	miR-21-5p	miR-25-3p	miR-28-5p	miR-34a-5p	miR-99a-5p	miR-122a-5p	miR-124a-3p	miR-125a-5p	miR-125b-5p
miR-126-3p	miR-131	miR-133a-3p	miR-133b	miR-143-3p	miR-145-5p	miR-146a-5p	miR-146b-5p	miR-148a-3p	miR-155-5p
miR-181a-5p	miR-181b-5p	miR-181c-5p	miR-182-5p	miR-192-5p	miR-194-5p	miR-195-5p	miR-199a-5p	miR-199b-5p	miR-199a*-3p
miR-200a-3p	miR-200c-3p	miR-204-5p	miR-206	miR-216-5p	miR-223-3p	miR-224-5p	miR-342-3p	mir-376c-3p	miR-375
miR-9-1-5p	miR-10b-5p	miR-17-3p	miR-22-3p	miR-23a-3p	miR-24-3p	miR-26a-5p	miR-26b-5p	miR-27a-3p	miR-27b-3p
miR-29a-3p	miR-29b-3p	miR-29c-3p	miR-30a-3p	miR-30a-5p	miR-30b-5p	miR-30c-5p	miR-92-3p	miR-92b-3p	miR-93-5p
miR-95-3p	miR-101-1-3p	miR-103-3p	miR-106a-5p	miR-106b-5p	miR-107	miR-128a-3p	miR-128b-3p	miR-132-3p	miR-134-5p
miR-135b-5p	miR-136-5p	miR-137	miR-140-5p	miR-141-3p	miR-142-3p	miR-149-5p	miR-150-5p	miR-151-3p	miR-153-3p
miR-154-5p	miR-181d-5p	miR-183-5p	miR-185-5p	miR-186-5p	miR-188-5p	miR-190-5p	miR-191-5p	miR-196a-5p	miR-196b-5p
miR-197-3p	miR-198	miR-200b-3p	miR-202-5p	miR-203a	miR-205-5p	miR-210-3p	miR-214-3p	miR-215-5p	miR-218-5p
miR-219	miR-221-3p	miR-222-3p	miR-296-5p	miR-372-3p	miR-373-5p	miR-488-5p	miR-100-5p	miR-127	miR-142-5p
miR-31-5p	miR-181a-3p	U6

Principle

miRNA is different from large messenger RNA in three aspects; (1) miRNAs are small size molecules with quite a big difference in abundance, (2) mature miRNAs co-exist with their precursor pre-miRNA and pri-miRNA, which only differ in length, and (3) many miRNAs are very closely related in sequences, such as isoforms, only differing by one or a few nucleotides. Therefore, the conventional mircoarray technologies cannot directly be applied to analyzing these molecules. A number of miRNA microarray products are commercially available, but they are either tedious in requiring pre-isolation of microRNA, lack the discriminative power to differentiate between isoforms, or are not sensitive enough to monitor low abundant miRNAs.

In our array assay, each miRNA molecule is targeted by two oligos that hybridize with the target miRNA to form a RNA/DNA duplex. When the sequences are perfectly matched, the oligos are aligned with the miRNA and the joint can be ligated by DNA ligase (figure 1). A single nucleotide difference among miRNAs will block either the hybridization or the ligation; Thus, miRNA isoforms can be differentiated. Due to the small size of miRNA, the hybrid might not be stable; Therefore, we introduce the stacking sequences. By extending these two oligos along with their complementary oligos, the stability is increased. Once the pair of oligos is ligated, the ligated molecules are subjected to linear amplification via T7 transcription into RNA in the presence of biotin-UTP, which are used as probes for array hybridization. To differentiate each isoform, we assigned unique tag sequences to the ligation oligos, so that single nucleotide differences are converted into unique tag sequences. In this way, each isoform can be easily distinguished by array hybridization.

The assay procedure includes three steps: (1) mix the miRNA with provided oligos to form miRNA/oligo hybrids; (2) select the hybrids and remove free oligos, and ligate miRNA-directed pairing of oligos to become a single DNA; and (3) amplify the ligated DNA with T7 transcription.

Data

Human miRNA array III analysis of miRNA expression in HeGP2, HEK293, and HeLa cells.

Literature

View user manual

Citations

1.  MicroRNA27a Regulates Cardiomyocytic Apoptosis During Cardioplegia-Induced Cardiac Arrest by TargetingInterleukin 10-Related Pathways.
 
2. Yeh CH, Chen TP, Wang YC, Lin YM, Fang SW. Shock. 2012 Dec;38(6):607-14.
 
3.  MTORC1-Dependent Dysregulated Expression Of MicroRNA Contributes To The Pathogenesis Of Lymphangioleiomyomatosis (LAM) Trindade AJ, Medvetz DA,  Neuman NA, Myachina F, Zhang E, Yu J, Henske EP. Am. J. Respir. Crit. Care Med., 2012 May; 185: A6821.
 
4. Curcumin inhibits the proliferation and invasion of human osteosarcoma cell line MG-63 by regulating miR-138, D Yu, F An, X He, X Cao. International Journal of Clinical and Experimental  2015. Int J Clin Exp Pathol. 2015; 8(11): 14946–14952. Published online 2015 Nov 1.