Exosome-based strategy against colon cancer using small interfering RNA-loaded vesicles targeting soluble a proliferation-inducing ligand

Figure 1 Generation of small interfering peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 RNA-loaded soluble a proliferation-inducing ligand-targeted exosome. A: Generation of small interfering peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 RNA (siPIN1)-loaded soluble a proliferation-inducing ligand (sAPRIL)-targeted exosome (tEx[p]). Briefly, the DNA sequence of the sAPRIL-targeting peptide was integrated into a pDisplay vector and used to transfect the adipose-derived stem cell (ASC), leading to the expression of the sAPRIL peptide on the cell membranes and the secretion of tEx. Subsequently, sIPIN1 was incapsulated into tEx using Exofect kit; B: Nanoparticle analysis of sAPRIL-targeted Ex. Zetaview analysis revealed that tEx had an average size of 187.9 ± 105.3 nm, aligning with the typical size range of exosomes (Ex), as corroborated by the transmission electron microscopy images; C: Western blot analysis showing PIN1 expression in various colon cancer cells. The increased expression of PIN1 was noted in HCT116 and SW620 colon cancer cells; D: Flow cytometric analysis of tEx. The tEx showed expression of markers CD63 and CD81 similar to control levels, while specifically expressing the myc marker (tEx marker) at a rate of 61.3% to 78.1%. Values are presented as mean ± standard deviation of three independent experiments. ^aP < 0.05, ^bP < 0.01. PIN1: Peptidyl-prolyl cis-trans isomerase NIMA-interacting 1.