The Effect of siRNA Transfection on the ARID3A and ERIC3A Gene Expression in Various Cell Lines Invitro

Abstract

Background: Thousands of distinct long non-coding RNAs are encoded in the human genome, and it is becoming clear that these transcripts play a crucial role in controlling gene expression and cell destiny. However, the transcriptional control of their expression is not well understood. The long noncoding RNA E2f1-regulated inhibitor of cell death (ERICD) is essential for inducing both cell death and proliferation and is a critical downstream target of the tumor suppressor. However, E2F-dependent transcription is triggered by ARID3A's binding to the E2F transcription factor. Aim of the study: Detection of the expression level of ARID3A and ERICD by transfection of siRNA and comparison with normal and negative control. Material and methods: Osteosarcoma cells (U2OS) were performed for ERICD and ARID3A expression. Experiments including overexpression and knockdown of ARID3A and a knockdown of ERICD were conducted. Assays for colony development and migration were also carried out in U2OS. The siRNA was used for the inhibition of both ERICD and ARID3A in U2OS. Extraction of mRNA from U2OS then converting to cDNA by using random primer then qPCR was used for detection of bands. Results: This investigation demonstrated that is the indirect pathway through which ARID3A and ERICD interact. In addition, ERICD and ARID3A exhibit carcinogenic properties in osteosarcoma. The discovery of a unique interaction between the ERICD and ARID3A marks a significant advancement in the understanding of lncRNAs that target DNA-binding proteins. The siRNA-mediated knockdown of ARID3A and ERICD significantly decreased colony formation and hindered cell migration in U2OS. Conversely, overexpression of ARID3A verified a noteworthy enhancement in wound closure and heightened capacity of U2OS cells to form colonies. Conclusions: ARID3A and ERICD have roles in transcription factors for the expression of cancer cells. During the transfection of siRNA, both genes inhibited cancer cells. ERICD when compared with non-transfected normal cells showed a significant decrease in the level.

Keywords

• ERICD
• ARID3A
• Overexpression
• Knockdown