Functional Study of Interferon Gamma Inducible Protein 16(IFI16)in Multiple Myeloma
Objective:To reveal the function of interferon gamma inducible protein 16(IFI16)in multiple myeloma(MM).Methods:Human multiple myeloma cells(RPMI-8266)were divided into Control group(untransfected cells),NC-sh group(cells transfected with negative control shRNA lentivirus),IFI16-sh group(cells transfected with IFI16 shRNA lentivirus),NC-OE group(cells transfected with negative control overexpressing lentivirus)and IFI16-OE group(cells transfected with IFI 16 overexpression lentivirus).Lipofectamine 2000 was used for transfection and cells were collected after 48 h culture.Cell proliferation was detected by MTT assay and colony for-mation test.Apoptosis was detected by Annexin V-FITC and PI staining.Cell invasion was detected by Transwell.The mRNA levels of IFI16,Bcl-2,Bax,fibroblast growth factor(FGF)1 and FGF2 were detected by qRT-PCR.The protein expression level of IFI16 was de-tected by Western blot.Nude mice were randomly divided into NC-sh group and IFI 16-sh group(n=6).Nude mice in NC-sh group and IFI16-sh group were subcutaneously injected with RPMI-8266 cell suspension transfected with NC-sh and IFI16-sh,respectively.After 35 days,the tumor volume was measured and the tumor weight was weighed.The levels of Th1 cytokines[interferon-γ(IFN-y),inter-leukin(IL)-12],Th2 cytokines(IL-4)and proinflammatory factors[IL-1β,IL-6,tumor necrosis factor-α(TNF-α)]were detected by ELISA method.Results:Compared with Control group and NC-sh group,the relative expression of IFI 16 mRNA and protein of RP-MI-8266 cells in IFI 16-sh group decreased(P＜0.05),relative cell viability and number of colony and invasive cells decreased(P＜0.05),the apoptosis rate and relative expression of Bax mRNA increased(P＜0.05),the relative mRNA expression level of Bcl-2,FGF1 and FGF2 decreased(P＜0.05).Compared with Control group and NC-OE group,the relative expression of IFI 16 mRNA and protein of RP-MI-8266 cells in IFI16-OE group increased(P＜0.05),relative cell viability and number of colony invasive cells increased(P＜0.05),apop-tosis rate and relative expression of Bax mRNA decreased(P＜0.05),the relative mRNA expression level of Bcl-2,FGF1 and FGF2 in-creased(P＜0.05).Compared with NC-sh group,the tumor volume,tumor weight,the relative expression of IFI16 mRNA and protein in tumor tissue of IFI16-sh group decreased(P＜0.05),the levels of IFN-γ and IL-12 in peripheral blood increased(P＜0.05),the level of IL-4,IL-1 β,IL-6 and TNF-α in peripheral blood decreased(P＜0.05).Conclusion:IFI 16 is a kind of oncogene in MM.Down-regulation of IFI16 can inhibit the growth and invasion of MM cells.The mechanism is related to correcting the balance of Thl/Th2 cells and inhibiting the activity of pro-inflammatory factors.
Interferon gamma inducible protein 16Multiple myelomaOncogeneThl/Th2 cell balancePro-inflammatory factor
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