The CRISPR/Cas system inhibited the pro-oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary XXXX cells.

@entity1 @entity5 @entity2347 @entity3 @entity22577
OBJECTIVES: To identify the association of fibronectin (FN) extra domain A ( @entity22577 ) with the progression of salivary @entity2347 (SACC). Accordingly, the exclusion of @entity22577 exon through the CRISPR/Cas9 system was investigated as the rescue for such pro-oncogenic splicing. MATERIALS AND METHODS: SACC-83 cells were transiently transfected with plasmids containing recombinant @entity22577 , and the cellular growth and motility were then accessed in vitro. Epithelial-mesenchymal transition (EMT) was investigated with immunohistochemistry, Western blot, and real-time PCR analysis. SACC tissues from 81 @entity1 were used to access the associations between @entity22577 +FN and clinical-pathological parameters. CRISPR/Cas9 plasmids containing sgRNA were designed and co-transfected into SACC-83 cells; the effects of @entity22577 knockout on cellular growth and motility were then accessed. RESULTS: The recombinant @entity22577 exhibited little effect on the proliferation of SACC cells, but significantly promoted the migration and invasion of the cells (P  <  0.05), accompanied with upregulated EMT (P  <  0.05); consistently, the expression of @entity22577 +FN was positively associated with the @entity3 , nerve invasion and recurrence of SACC (P  <  0.05). Furthermore, the @entity22577 knockout from the FN gene in most SACC cells resulted in a decrease in cell motility and invasion, as well as prolonged population doubling time, compared with untreated SACC-83 cells (P  <  0.05). CONCLUSION: The @entity22577 domain significantly promoted the motility of SACC cells, and positively associated with the @entity5 progression in @entity1 with SACC. Thus, it is a potential risk factor and also a therapeutic target for SACC. The CRISPR/Cas9 system may control a pro-oncogenic splicing process through the exclusion of @entity22577 exon from the FN gene, leading to inhibition of motility, invasion and proliferation of @entity5 cells.
