Protein profiling analysis of oral squamous cancer derived extracellular vesicle subgroups

doi:10.19438/j.cjoms.2023.04.002

Abstract

Abstract: PURPOSE: To reveal the commonality and heterogeneity in protein contents of extracellular vesicles (EVs) subgroups obtained by different immunomarkers, and provide data support for researchers to select appropriate isolation protocols. METHODS: In this study, oral cancer cells CAL27, pancreatic cancer cells PANC-1 and gastric cancer cells AGS were used, and EVs derived from them were isolated by 4 EVs markers CD9, CD63, CD81 and EpCAM. Then the proteins from these 4 different EVs subgroups were detected by mass spectrometry. The commonality and heterogeneity between EVs subgroups were evaluated by funrich3.1.3 software. SPSS 19.0 software package was used for statistical analysis. RESULTS: Under the same culture conditions, AGS cells produced the highest number of EVs, followed by CAL27 cells and the least by PANC-1 cells. Among the 4 different EVs isolation methods, the EpCAM marker-based isolation method was the most efficacious. The results of principal component analysis showed that the subgroups of EVs involved in this study were heterogeneous at the level of cell origin as well as at the level of isolation markers. The results of the cellular component analysis of the protein profile showed that 86.11% of the proteins enriched by CD63⁺ EVs derived from CAL27 were related to exosomes, which was significantly higher than the percentage of the CD9⁺, CD81⁺ and EpCAM⁺ EVs subgroups(70.92%, 70.90% and 63.68%); while 37.96% of the enriched proteins in the CD63⁺ EVs subgroup were related with cell membrane, which was lower than the percentage of CD9⁺, CD81⁺, and EpCAM⁺ EVs subgroups (53.20%, 61.19% and 49.53%), indicating that EVs subgroups isolated by different immunomarkers had different origins. CONCLUSIONS: The subgroups of EVs involved in this study are heterogeneous at the cellular origin level and at the immunomarker level. Among them, CD63⁺ EVs may originate more from the endosomal pathway, while CD9⁺, CD81⁺ and EpCAM⁺ EVs originate more from the plasma membrane outgrowth pathway. The EVs subgroups obtained based on different isolation markers were differential in terms of cell biological functions.

Key words: Extracellular vesicles, Tumor, Heterogeneity, Isolation markers, Ectosome

CLC Number:

R739.8

WU Ruo-yi, WANG Xiao-ning, ZHAI Pei-song, ZHANG Jian-jun, CHEN Wan-tao. Protein profiling analysis of oral squamous cancer derived extracellular vesicle subgroups[J]. China Journal of Oral and Maxillofacial Surgery, 2023, 21(4): 326-331.

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