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siRNA induces gelsolin gene transcription activation in human esophageal cancer cell
Guo-Wei Huang1, Lian-Di Liao2, En-Min Li3
11] Institute of Oncologic Pathology, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China [2] The Key Laboratory of Molecular Biology for High Cancer Incidence Coastal Chaoshan Area, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China [3] Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China.
Abstract:
Recent studies show that targeting gene promoter or 3' terminal regions of mRNA with siRNA induces target gene transcription. However, the ability of exon-targeting siRNA to affect transcription has yet to be demonstrated. We designed and synthesized siRNA against various exons in the gelsolin gene (GSN) to knockdown GSN transcript in KYSE150 and KYSE450 cells. Surprisingly, we found that siGSN-2, targeting the GSN twelfth exon, induced GSN gene transcription detected by real time RT-PCR. An siGSN-2 co-precipitation assay was performed and H3 histone, previously shown to correlate with gene transcription, was detected in the siGSN-2 pull-down pellet. However, H3 histone was not detected in an siGSN-1-precipitated pellet, which resulted in GSN knockdown. In addition, siGSN-2 decreased stress fibers, lamellipodia and filopodia, demonstrating that siGSN-2 induced GSN transcription activation and exerted biological function. In conclusion, our finds reveal siRNA, which is derived from target gene exon, can form the complex with H3 histone to be involved in the regulation of gene expression.
