Antitumor Activity Of The Protein Of Aprionaswainsoni Larvae In Vitro And Its Immunoregulatory Effect On Mouse Macrophages

Study the anti-tumor activity and immunomodulatory effects of the protein of Aprionaswainsoni larvae in vitro.The total protein of Aprionaswainsoni 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 Aprionaswainsoni 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 proliferation.Migration assay was used to detect the effects of the protein of Aprionaswainsoni larvae on the migration ability of 4T1 cells.Flow cytometry was used to study the effect of the protein of Aprionaswainsoni larvae on apoptosis of MFC,4T1 cells.Fluorescence microspheres assay for the effect of the the protein of Aprionaswainsoni larvae on phagocytic ability of RAW264.7 cells.The production of NO(nitric oxide)production was evaluated by Griess.The production of TNF-?(prostaglandin tumor necrosis factor),IL-6(interleukin-6),and IL-1(interleukin-1?)were determined by ELISA(immunosorbent assay).The mRNA levels of TNF-?(tumor necrosis factor),IL-1(interleukin-1),TLR 4(toll-like receptors4),MIR-7(mirn7),IFN-?(interferon-?),IL-6(interleukin-6)associated with cell phagocytic ability in RAW264.7 cells were detected by qRT-PCR.The mRNA levels of MMP2(matrix metalloprotein 2),MMP9(matrix metalloprotein 9),signal transducer and Stat3(activators of transcription 3),c-Myc,andSdf1(stromalcell-derived factor-1)associated with cell proliferation and migration in 4T1 cells were detected by qRT-PCRCompared with normal control group,the protein of Aprionaswainsoni larvae showed a good inhibitory effect on the proliferation of MFC and 4T1 cells.The inhibition rate of MFC cell proliferation can reach 83%,and the inhibition rate of 4T1 cell proliferation reaches 53%.The protein 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 normal control group,suggesting that the protein of Aprionaswainsoni larvae inhibited the migration of 4T1 cells.The protein of Aprionaswainsoni larvae could induce apoptosis in MFC and 4T1 cells(P<0.05).The protein of Aprionaswainsoni larvae significantly increased phagocytic ability and production of NO,TNF-?,IL-6 and IL-1?.The mRNA levels of TNF-?,IL-1,TLR4,MIR-7,IFN-? 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 Sdfl in 4T1 cells of the test groups were significantly upregulated(P<0.05,P<0.01).The protein of Aprionaswainsoni larvae has good in vitro antitumor activity and potential immunomodulatory effects.