Gain-of-Function p53 Protein Transferred via Small Extracellular Vesicles Promotes Conversion of Fibroblasts to a Cancer-Associated Phenotype

Crosstalk between tumor and stromal cells is essential for tumor pathogenesis. Tumor and stromal interactions consist of reciprocal signaling through cytokines, growth factors, direct cell-cell interactions, and extracellular vesicles (EVs), among others. Small EVs (=200 nm) have been considered critical messengers of cellular communication during tumor development. Here, we demonstrated that gain-of-function (GOF) p53 protein can be packaged into small EVs and transferred to stromal cells. In this study, we performed RNA sequencing to explore the gene signature alterations in small EVs after knockdown GOF p53 expression. Overall design: Small EVs were isolated from HT29-nontargeting control sequence (ntsh) cells (ntsh-sEVs) and HT29-p53 shRNA cells (shp53-sEVs). RNA samples from small EVs were extracted using an exoRNeasy Midi and Maxi Kit (Qiagen, Hilden, Germany; Cat. # 77144) following the manufacturer's instructions. The library construction for small RNA and the sequencing were performed by Novogene Corporation Inc. (Sacramento, CA).