The transcriptome analysis of cleft lip/palate-related PTCH1 variants in GMSM-K cells show carcinogenic potential

[摘要] Cancer progression involves the sonic hedgehog (SHH) pathway, in which the receptor PTCH1 actives thedownstream pathways. Dysfunction of PTCH1 can lead to nevoid basal cell carcinoma Syndrome (NBCCs) includingneoplastic disease and congenital disorder. To evaluate the relationship between PTCH1 and cancer, we applied theCRISPR/Cas9 system to knock out PTCH1 in oral nontumorous epithelial cells (GMSM-K). Then we screened sixPTCH1 variants associated with cleft lip/palate (CL/P), one of the congenital disorders in NBCCs, and generatedPTCH1 variant and wild-type recombinant PTCH1−/− GMSM-K cell lines. Transcriptome sequencing was conductedin these cell lines. The results revealed that differentially expressed genes (DEGs) in PTCH1−/− GMSM-K wereenriched in extracellular compartments, contributing epithelial diseases by pathway enrichment analysis. RT-PCRconfirmed that KRT34, KRT81, KRT86, PDGFB, and WNT10B genes, associated with extracellular compartmentswere highly expressed in PTCH1−/−. The Kyoto Encyclopedia of Genes and Genomes analysis also suggested thatDEGs are closely related to focal adhesion, transcriptional misregulation, and proteoglycans in breast and gastriccancers. Comparative analysis of samples revealed that the CL/P-associated PTCH1 variants A443G and V908G arepotentially carcinogenic. These findings provide new insights into the carcinogenic potential of PTCH1 dysfunction.

[学科分类] 仪器

[关键词] CRISPR/Cas9;PTCH1 variant;Carcinogenic potential;Transcriptome sequencing