Therapeutic Efficacy Analysis of lncRNA NEAT1 Gene Knockout and Apoptosis Induction in Prostate Cancer Cell Line Using CRISPR/Cas9

Abstract

Background and Objective: Long non-coding ribonucleic acid (lncRNA) has been identified as an important gene regulator and prognostic marker in various cancers. The present study aimed to investigate the effects of Nuclear Paraspeckle Assembly Transcript1 (NEAT1) gene knockout using Clustered Regularly Interspaced Short Palindromic Repeats-associated Protein 9 (CRISPR/Cas9) in PC-3 cell line. Materials and Methods: In this experimental study, recombinant plasmids carrying single guide RNA in the CRISPR system were designed and constructed. The prostate cancer PC3 cell lines were transfected by recombinant vectors, and apoptotic gene expression was evaluated using the quantitative polymerase chain reaction technique. Furthermore, annexin, MTT, and wound healing assays were applied for apoptosis, cell proliferation, and cell migration activities respectively. Results: Knockout of NEAT1 gene using CRISPR/Cas9 technique caused significant changes in the expression of apoptosis-related genes in a way that the expression of P21, BCL2, and BIRC5 genes decreased, while the expression of P53 and BAX increased. In addition, in the treatment group, the rate of cell proliferation and migration was reduced compared to those in the control group. Moreover, an increase was observed in the apoptosis rate in genetically modified cells compared to the control group. Conclusion: As an oncogene, NEAT1 plays an important role in tumorigenesis and its knockout reduces cell survival and migration and increases the apoptosis of cancer cells.

Keywords

• crispr-associated protein 9
• neat1 long non-coding rna
• prostatic neoplasms
• pc-3 cells