Abstract

To explore new therapeutic options for cervical cancer, the inhibitory effect on cervical cancer of targeted CD133-loaded sPD1 gene microbubbles (MBs) combined with low-frequency ultrasound was studied and its mechanism was explored. We prepared microbubbles conjugated with anti-CD133 antibody to deliver the sPD1 gene and determined concentration, particle size, and potentials of MBs. In addition, we verified that CD133 targeted-MBs could specifically bind to U14 cervical cancer cells in vitro. A mouse model of subcutaneous xenograft cervical cancer was established and mice were divided into a control group, an non-targeted microbubble group, a CD133-MBs group, an sPD1-MBs group and a CD133/sPD1-MBs group. Compared with the control group, tumor growth was inhibited in each group, with the CD133/sPD1 group showing the strongest inhibitory effect after treatment. The tumor volume and weight inhibition rates in the CD133/sPD1-MBs group were 78.01% and 72.25% respectively, which were statistically different from the other groups (P < 0.05), and HE staining and TUNEL immunofluorescence showed necrosis and apoptosis in tumor tissue. Flow cytometry, lactate dehydrogenase, and indirect immunofluorescence experiments showed that T lymphocytes were activated and a large number of CD8-positive T cells infiltrated the tumor tissue after treatment, with the CD133/sPD1-MBs group showing the most prominent effects (P < 0.05). The combination of ultrasound with anti- CD133 antibody-conjugated microbubbles loaded with the sPD1 gene can inhibit the growth of cervical cancer, suggesting that the immunosuppressive microenvironment of the tumor is improved after treatment.

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