apoVs from HSC-3 cells transfer CCT2 to SAS cells and promote their cisplatin resistance (IMAGE)

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apoVs from HSC-3 cells transfer CCT2 to SAS cells and promote their cisplatin resistance

Caption

(A) HSC-3 control and shCCT2 cells were treated with 20 μM cisplatin for 48 h to induce apoptosis. The apoptotic cell suspensions were then isolated using a sequential centrifugation system to obtain apoVs. (B) Nanoflow cytometry analysis showed the annexin V-positive rate of apoVs. (C) Nanoparticle tracking analysis confirmed the size distribution of apoVs. (D) Transmission electron microscopy assay showed the morphology of apoVs. Scale bar: 200 nm. (E) Western blotting revealed the expression of lamin B1, FAS, RhoA, and syntenin-1 in aopVs and exosomes. (F) Western blot images of CCT2 expression in apoVs derived from control and shCCT2 HSC-3 cells. (G) Nanoflow cytometry analysis showed the positive rate of CCT2 in apoVs. (H) Immunofluorescence images showed that the apoVs were internalized by receptor-tumor cells. Scale bar: 10 μm. (I) Flow cytometry analysis showed the apoptosis levels of SAS cells after uptake of apoVs from HSC-3 control and shCCT2 cells, followed by treatment with different concentrations of cisplatin for 2 days. (J) CCK-8 assay evaluated the effect of apoVs on the cisplatin sensitivity of SAS cells. (K) BrdU assay confirmed that apoVs influenced the cell cycle of SAS cells. (L) Western blotting assay showed that apoVs affected the expression levels of CCT2, c-Myc, CCND1, and CCNE1 proteins in SAS cells. (M) Schematic diagram of CCT2 stabilizing c-Myc to drive tumor progression and chemoresistance in HNSCC, with vesicle-mediated transfer promoting drug resistance in recipient cells. Data were presented as mean ± standard deviation. ns (not significant), ∗p < 0.05 and ∗∗∗p < 0.001 for student's t-test and one-way ANOVA test. apoVs, apoptotic vesicles; CCT2, chaperonin containing TCP1 subunit 2; CCND1, cyclin D1; CCNE1, cyclin E1; HNSCC, head and neck squamous cell carcinoma.

Credit

Lin Lu, Yi Shen, Xuerong Li, Yiwei Zhao, Xuefan Zhai, Min Cai, Baicheng Bao, Guiqing Liao, Jianbo Sun

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