| Objective: To investigate the effect of ultrasound irradiation on the proliferation and migration of ovarian cancer cells with CXCR4 antibody and PD-L1 antibody double targeted nanobubbles.Methods: IOSE-80 normal ovarian epithelial cells,SKOV3 and CAOV3 ovarian cancer cells were cultured and expanded.Double labeling fluorescence immunoassay was used to co-locate CXCR4 and PD-L1 proteins.Western blot was used to detect the relative expression of CXCR4 and PD-L1 proteins in three kinds of cells and screen out the experimental cells.The experimental cells were co-incubated with targeted nano-bubbles with different ligands and irradiated by ultrasound.The experimental cells were divided into 6 groups according to different modes of action.CCK8 test and wound healing test were used to compare the ability of cell proliferation and migration.Results:(1)Double-labeled fluorescence immune image assay showed that the cell membrane expressing CXCR4 was dyed green,while the cell membrane and cytoplasm expressing PD-L1 dyed red.CXCR4 and PD-L1 proteins could be expressed in all three kinds of cells.(2)Western blot showed that the expression of CXCR4 and PD-L1 in two kinds of cancer cells was significantly higher than that in IOSE-80 cells(P<0.05).There was no significant difference between the two kinds of cancer cells(P>0.05).Therefore,SKOV3 cancer cells were selected as the follow-up experimental cells.(3)In the cancer cell experiment,the cell survival rate and healing rate of the double targeted nanobubble group were(28.6±3.1)% and(16.5±3.4)%,respectively,which were significantly different from those of the single targeted nanobubble group(P<0.05).Conclusion:The results showed that the use of CXCR4 antibody and PD-L1 antibody double targeted nanobubbles under UTMD can significantly inhibit the proliferation and migration of ovarian cancer cells,providing strong evidence that combined immunotherapy can enhance the anti-tumor effect. |
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