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Archive > Volume 29 Issue 4 > 2022,29(4):284-293. DOI:10.3872/j.issn.1007-385X.2022.04.003 Prev Next
Prompt Report
Anti-HSP90 antibody promotes the anticancer effect of cisplatin by inhibiting human gastric cancer stem cells through internalizing eHSP90
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Abstract:
Objective: To explore the promotive effect of anti-HSP90 monoclonal antibody mAb 28C10 on Cisplatin (DDP) inhibiting malignant biological behavior of human gastric cancer PAMC82 cells by targeting tumor stem cells and the possible mechanisms of action. Methods: Human gastric cancer PAMC82 cells were treated with 28C10 alone or in combination with DDP. Then, the serum- free pellet-forming ability, clone-forming ability, migration and invasion ability of PAMC82 cells were detected by different experiments, and the effects of 28C10 on the malignant biological behaviors of PAMC82 cells and the synergistic anti-cancer effect of 28C10 and DDP were detected by CCK-8 method. The expression, localization, eHSP90 + subgroup ratio of HSP90 and eHSP90 (extracellular HSP90) in PAMC82 cells and the effects of 28C10 on ALDH + , CD44 + , eHSP90 + subgroups of PAMC82 cells were detected by cellular immunofluorescence and flow cytometry. The changes in HSP90, stemness-related proteins and PI3K/AKT/mTOR signaling pathway in PAMC82 cells treated with 28C10 were detected by WB. Results: eHSP90 was expressed on the membrane surface of gastric cancer PAMC82 cells, and there were about 2%-3% of eHSP90 + subgroup cells. eHSP90 + cells were mostly co-positive with ALDH + or CD44 + cells. 28C10 significantly inhibited the ability of pellet formation, clone formation, proliferation, drug resistance, migration and invasion of PAMC82 cells; moreover, the effect was more obvious when combined with DDP (all P<0.05 or P<0.01). Flow cytometry analysis showed that 28C10 treatment significantly inhibited the number of eHSP90 + , ALDH + and CD44 + subgroups of PAMC82 cells (all P<0.01). Immunofluorescence assay showed that endocytosis of eHSP90 occurred after 28C10 treatment. WB results showed that the expression levels of eHSP90, CD44, ALDH and the expression of stemness-related proteins OCT4 and SOX2 were decreased (P<0.05 or P<0.01). Conclusion: 28C10 can target ALDH + and CD44 + tumor stem cell-related subgroups of gastric cancer PAMC82 cells, internalize eHSP90, reduce the level of total HSP90, and inhibit PI3K/AKT/mTOR signaling pathway, thereby effectively inhibiting stemness, drug resistance and other malignant biological behaviors of PAMC82 cells and synergistically improving the anticancer effect of DDP.
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Clc Number:
R735.2；R730.554；R730.53
