Anti-Tumor Activity of the Protein of Apriona swainsoni Larvae In Vitro and Its Immunoregulatory Effect on Mouse Macrophages

Here, we studied the anti-tumor activity and immunomodulatory effects of the protein of Apriona swainsoni larvae in vitro. The total protein of Aprionas wainsoni larvae was extracted and divided into three parts. The different protein sites were obtained after dialysis and salt removal. The molecular weights of different proteins of Apriona swainsoni larvae were detected by SDS-PAGE. The MTT method was used to study the human gastric cancer MFC, the mouse breast cancer 4T1 cells, macrophage RAW264.7 cells and the human umbilical endothelialvein HUEVC cells proliferation. Migration assay was used to detect the effects of the protein of Apriona swainsoni larvae on the migration ability of 4T1 cells. Flow cytometry was used to study the effect of the protein of Apriona swainsoni larvae on apoptosis of MFC, 4T1 cells. Fluorescence microspheres assay for the effect of the the protein of Apriona swainsoni larvae on phagocytic ability of RAW264.7 cells.The production of NO (nitric oxide) production was evaluated by Griess. The production of TNF-α (tumor necrosis factor-α), IL-6 (interleukin-6), and IL-1 (interleukin-1) were determined by ELISA (immunosorbent assay). The mRNA levels of TNF-α (tumor necrosis factor-α), IL-1 (interleukin-1), TLR4 (toll-like receptors 4), MIR-7 (mirn-7), IFN-γ (interferon-γ), IL-6 (interleukin-6) associated with cellphagocytic ability in RAW264.7 cells were detected by RT-PCR. The mRNA levels of MMP2 (matrix metalloprotein 2), MMP9 (matrix metalloprotein 9), signal transducer and STAT3 (signal transducer and activators of transcription 3), c-Myc and Sdf1 (stromal cell-derived factor 1) associated with cell proliferation and migration in 4T1 cells were detected by RT-PCR. Compared with the normal group, the protein of Apriona swainsoni larvae showed a good inhibitory effect on the proliferation of MFC and 4T1 cells, and had no effect on the proliferation of RAW264.7 cells (P<0.05). The cell migration rates of test groups were reduced (P<0.01) when compared with the normal group, suggesting that the protein of Apriona swainsoni larvae inhibited the migration of 4T1 cells. The protein of Apriona swainsoni larvae could induce apoptosis in MFC and 4T1 cells (P<0.05), and have no cytotoxic effect on HUEVC cells. The protein of Apriona swainsoni larvae significantly increased phagocytic ability and production of NO, TNF-α, IL-6 and IL-1β. The mRNA levels of TNF-α, IL-1, TLR4, MIR-7, IFN-γ and IL-6 in RAW264.7 cells of the test groups were significantly upregulated (P<0.05, P<0.01). The mRNA levels of MMP2, MMP9, STAT3, c-Myc and Sdf1 in 4T1 cells of the test groups were significantly upregulated (P<0.05, P<0.01). The protein of Apriona swainsoni larvae has good in vitro antitumor activity and potential immunomodulatory effects.

CSTR: 32200.14.cjcb.2020.02.0012