Targeting the MALAT1 gene with the CRISPR/Cas9 technique in prostate cancer

Abstract

Background The MALAT1 lncRNA acts as an oncogene in Prostate cancer (PC); thus, it can be severe as a cancer biomarker. Methods Using bioinformatics datasets including (HTSeq-Counts, GDC, and TCGA) 5501 gene expression profiling specimens were gathered. Then, expression profiles and sample survival of lncRNA were investigated using COX regression analyses, ROC curve analysis. The Database for Annotation, Visualization, and Integrated Discovery was used to conduct GO and KEGG studies on the lncRNA-related PCGs. After MALAT1 Knockout via CRISPR/Cas9 technique, the MALAT1 expression was assessed in DU-145 cells. The deletion of the target fragment was examined by polymerase chain reaction (PCR). Also, the expression of apoptosis genes was investigated by qRT-PCR. The viability and cell proliferation were measured using the MTT assay. Cell migration capability was determined using the cell scratch assay. The results of qRT-PCR were assessed by the ΔΔCt method, and finally, statistical analysis was performed in SPSS software. Results A maximum of 451 lncRNAs were discovered to reflect different expressions between PC and non-carcinoma tissue samples, with 307 being upregulated and 144 being down-regulated. Thirty-six lncRNAs related to OS were carefully selected, which were then subjected to stepwise multivariate Cox regression analysis, with 2 lncRNAs (MALAT1, HOXB-AS3). MALAT1 is highly expressed in PC cells. MALAT1 Knockout in DU-145 cells increases apoptosis and prevents proliferation and migration, and DU-145 transfected cells were unable to migrate based on the scratch recovery test. Overall, data suggest that MALAT1 overexpression in PC helps metastasis and tumorigenesis. Also, MALAT1 knockout can be considered a therapeutic and diagnostic target in PC. Conclusion Targeting MALAT1 by CRISPR/Cas9 technique inhibit the cell proliferation and migration, and in addition induce apoptosis. Thus, MALAT1 can act as a tumor biomarker and therapeutic target.

The Expanding Roles of Long Non‐Coding RNAs in Pancreatic Cancer

“…[ 123 ] The CRISPR‐Cas9 technology can also be employed to induce double‐stranded DNA breaks in specific regions of lncRNAs. [ 124 ] Another method for targeting lncRNAs involves the use of antisense oligonucleotides (ASOs) to regulate lncRNA expression and function. ASOs are short synthetic oligonucleotide sequences that bind to complementary sequences of target lncRNAs.…”

Long noncoding RNA MALAT-1: A versatile regulator in cancer progression, metastasis, immunity, and therapeutic resistance

“…Multiple investigations have demonstrated a connection between increased MALAT-1 levels and tumor development, invasion, and metastasis in PCa. Further research into MALAT-1 is crucial since it has also been linked to promoting resistance to several anticancer treatments [ [133] , [134] , [135] , [136] ].…”

Long non-coding RNAs as critical regulators and novel targets in cervical cancer: current status and future perspectives

“…They have already been proposed as potential diagnostic and prognostic biomarkers in cancers of different sites including breast, lung, liver, prostate, cervical and pancreas [126][127][128][129][130]. LncRNAs PCA3 (Prostate cancer gene 3) and MALAT-1 (Metastasis associated lung adenocarcinoma transcript 1) show the potential to be used in clinical practices as diagnostic biomarkers for prostate cancer [131,132]. Interestingly, the lncRNAs possessing diagnostic potential are also found in the body fluids with satisfactory stability, thereby enabling a painless diagnosis for the patient.…”