Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD

Abstract

Purpose:RNA activation (RNAa) is a mechanism of gene activation triggered by promoter-targeted small double stranded RNAs (dsRNAs), also known as small activating RNAs (saRNAs). Myogenic regulatory factor MyoD is regarded as the master activator of myogenic differentiation cascade by binding to enhancer of muscle specific genes. Stress urinary incontinence (SUI) is a condition primarily resulted from urethral sphincter deficiency. It is thus expected that by promoting differentiation of adipose-derived stem cells (ADSCs) into myoblasts by activating MyoD gene through RNAa may offer benefits to SUI.Materials and Methods:Rats ADSCs were isolated, proliferated in vitro, and identified by flow cytometry. Purified ADSCs were then transfected with a MyoD saRNA or control transfected. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to detect MyoD mRNA and protein expression, respectively. Immunocytochemical staining was applied to determine the expression of desmin protein in transfected cells. Cell viability was measured by using CellTiter 96® AQueous One Solution Cell Proliferation Assay kit.Results:Transfection of a MyoD saRNA (dsMyoD) into ADSCs significantly induced the expression of MyoD at both the mRNA and protein levels, and inhibited cell proliferation. Desmin protein expression was detected in dsMyoD treated ADSCs 2 weeks later.Conclusion:Our findings show that RNAa mediated overexpression of MyoD can promote transdifferentiation of ADSCs into myoblasts and may help treat stress urinary incontinence (SUI)–a condition primarily resulted from urethral sphincter deficiency.

saKLK1-374 is More Difficult to Induce KLK1 Expression in Normal Cell Lines Than in Tumor Cell Lines And Inhibits the Growth of Prostate Cancer Cells Not via Induction of KLK1 Expression

“…Bin Wang activated SOX2 in human lung diploid broblast via saRNA carried by lentiviral vector 23 . Chenghe Wang induced the expression of myogenic regulatory factor in adiposederived stem cells via cationic-lipid transfection reagent RNAiMax which was consistent with the transfection reagent in our research 24 . Perhaps the virus could carry more exogenous saRNA into normal cells.…”

A Lipid-Nanosphere-Small MyoD Activating RNA-Bladder Acellular Matrix Graft Scaffold [NP(saMyoD)/BAMG] Facilitates Rat Injured Bladder Muscle Repair and Regeneration [NP(saMyoD)/BAMG]

“…MTT assay was performed to determine the toxic effects ofNP(saMyoD)/BAMG and SF (VEGF)/BAMG scaffolds on ADSCs. Comparing to the blank group, NP(saMyoD)/BAMG slightly inhibited the proliferation of ADSC, which is consistent with our previous study (Wang et al, 2015), but NP(saMyoD)/BAMG is not toxic accroding to the China national standards for medical devices evaluation (GB/T16886.5-2017), because the inhibition ratio is lower than 20%. The cell morphology is normal under the microscope (Figure 5C).…”