Trans-splicing repair of mutant p53 suppresses the growth of hepatocellular carcinoma cells in vitro and in vivo

Abstract

Reactivation of wild-type p53 (wt-p53) function is an attractive therapeutic approach to p53-defective cancers. An ideal p53-based gene therapy should restore wt-p53 production and reduces mutant p53 transcripts simultaneously. In this study, we described an alternative strategy named as trans-splicing that repaired mutant p53 transcripts in hepatocellular carcinoma (HCC) cells. The plasmids which encoded a pre-trans-splicing molecule (PTM) targeting intron 6 of p53 were constructed and then transfected into HCC cells carrying p53 mutation. Phenotypic changes of HCC cells induced by p53-PTM were analyzed through cell cycle, cell apoptosis and the expression of p53 downstream target genes. Spliceosome mediated RNA trans-splicing (SMaRT) reduced mutant p53 transcripts and produced functional wt-p53 protein after the delivery of p53-PTM plasmids, which resulted in phenotype correction of HCC cells. In tumor xenografts established by p53-mutated HCC cells, adenovirus encoding p53-PTM induced cell cycle arrest and apoptosis and then blocked the growth of tumors in mice. Collectively, our results demonstrated for the first time that mutant p53 transcripts were functionally corrected in p53-defective HCC cells and xenografts using trans-splicing, which indicated the feasibility of using trans-splicing to repair p53 mutation in p53-defective cancers.

Highly efficient delivery of potent anticancer iminoquinone derivative by multilayer hydrogel cubes

“…It is noteworthy that BA-TPQ-hydrogel cubes display excellent inhibitory effects in both p53 wide-type (HepG2) and p53-mutant (Huh7) liver cancer cells indicating a p53-independent antitumor activity. Since the p53 mutation commonly occurs in HCC which may promote tumorigenesis and is associated with poor therapeutic outcomes [81], BA-TPQ-hydrogel cubes can be a successful candidate for HCC treatment due to their p53-independent antitumor activity.…”

Propofol Inhibits Proliferation, Migration, Invasion and Promotes Apoptosis Through Down-Regulating miR-374a in Hepatocarcinoma Cell Lines

“…TP53 mutation is one of the common genetic alterations in HCC, suggesting the critical role of TP53 in HCC [43]. Restoration of TP53 has been reported to suppress growth of HCC in vitro and in vivo [44]. Therefore, the interaction between miR-374a and TP53 was further studied among those predicted targets.…”

Differential Impacts of Alternative Splicing Networks on Apoptosis

“…A spliceosome-mediated RNA trans- splicing (SMaRT) strategy was recently reported to correct the mutant TP53 gene to the wild-type TP53 gene through a trans- splicing mechanism which involves splicing between two individual transcripts [77]. In brief, the expression plasmid containing a pre- trans- spliced exon that encodes the correct TP53 fragment is delivered into TP53 -defective hepatocellular carcinoma (HCC) cells.…”

“…In brief, the expression plasmid containing a pre- trans- spliced exon that encodes the correct TP53 fragment is delivered into TP53 -defective hepatocellular carcinoma (HCC) cells. Mutant TP53 transcripts were corrected by replacing the mutant exon with the trans- spliced exon, which encoded the functional TP53 protein in TP53 -defective HCC cells [77]. Introduction of the pre- trans- spliced TP53 exon mediated activation of TP53-responsive genes and subsequently suppressed the progression of HCC cells in vitro.…”