| 高 静,严学倩,肖 方,及月茹,陈 怡,李瑗春.干扰素γ诱导蛋白16（IFI16）在多发性骨髓瘤中的功能研究[J].,2024,(6):1030-1037 |
| 干扰素γ诱导蛋白16（IFI16）在多发性骨髓瘤中的功能研究 |
| Functional Study of Interferon Gamma Inducible Protein 16 (IFI16) in Multiple Myeloma |
| 投稿时间：2023-11-27 修订日期：2023-12-22 |
| DOI：10.13241/j.cnki.pmb.2024.06.005 |
| 中文关键词: 干扰素γ诱导蛋白16 多发性骨髓瘤 致癌基因 Th1/Th2细胞平衡 促炎因子 |
| 英文关键词: Interferon gamma inducible protein 16 Multiple myeloma Oncogene Th1/Th2 cell balance Pro-inflammatory factor |
| 基金项目:陕西省自然科学基础研究计划项目（2020JQ-460） |
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| 中文摘要: |
| 摘要 目的：探究干扰素γ诱导蛋白16（IFI16）在多发性骨髓瘤（MM）中的生物学功能。方法：将人多发性骨髓瘤细胞（RPMI-8266）分为Control组（未转染的细胞）、NC-sh组（转染阴性对照shRNA慢病毒的细胞）、IFI16-sh组（转染IFI16 shRNA慢病毒的细胞）、NC-OE组（转染阴性对照过表达慢病毒的细胞）和IFI16-OE组（转染IFI16过表达慢病毒的细胞）。使用Lipofectamine 2000进行细胞转染，培养48 h后收集细胞。通过MTT法和集落形成实验检测细胞增殖。通过Annexin V-FITC和PI染色法检测细胞凋亡。通过Transwell检测细胞侵袭。通过qRT-PCR检测IFI16、Bcl-2、Bax、成纤维细胞生长因子（FGF）1、FGF2的mRNA水平。通过Western blot检测IFI16的蛋白表达水平。将裸鼠随机分为NC-sh组和IFI16-sh组，每组6只。NC-sh组和IFI16-sh组裸鼠分别皮下注射转染NC-sh和IFI16-sh的RPMI-8266细胞悬液。35 d后测量肿瘤体积和肿瘤重量。采用ELISA法检测Th1细胞因子[干扰素-γ（IFN-γ）、白介素（IL）-12]、Th2细胞因子（IL-4）和促炎因子[IL-1β、IL-6、肿瘤坏死因子-α（TNF-α）]的水平。结果：与Control组和NC-sh组比较，IFI16-sh组RPMI-8266细胞的IFI16 mRNA和蛋白相对表达量降低（P<0.05），相对细胞活力、集落数量和侵袭细胞数量均降低（P<0.05），细胞凋亡率和Bax mRNA相对表达量均升高（P<0.05），Bcl-2、FGF1和FGF2的mRNA相对表达量均降低（P<0.05）。与Control组和NC-OE组比较，IFI16-OE组RPMI-8266细胞的IFI16 mRNA和蛋白相对表达量升高（P<0.05），相对细胞活力、集落数量和侵袭细胞数量均升高（P<0.05），细胞凋亡率和Bax mRNA相对表达量均降低（P<0.05），Bcl-2、FGF1和FGF2的mRNA相对表达量均升高（P<0.05）。与NC-sh组比较，IFI16-sh组裸鼠的肿瘤体积、肿瘤重量以及肿瘤组织中IFI16的mRNA和蛋白相对表达量均降低（P<0.05），外周血IFN-γ和IL-12水平均升高（P<0.05），IL-4、IL-1β、IL-6和TNF-α水平均降低（P<0.05）。结论：IFI16是MM中的一种致癌基因，下调IFI16可抑制MM细胞的生长和侵袭，其机制与纠正Th1/Th2细胞平衡和抑制促炎因子活性有关。 |
| 英文摘要: |
| ABSTRACT 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 IFI16 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 formation 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 detected by Western blot. Nude mice were randomly divided into NC-sh group and IFI16-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-γ), interleukin (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 IFI16 mRNA and protein of RPMI-8266 cells in IFI16-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 IFI16 mRNA and protein of RPMI-8266 cells in IFI16-OE group increased (P<0.05), relative cell viability and number of colony invasive cells increased (P<0.05), apoptosis rate and relative expression of Bax mRNA decreased (P<0.05), the relative mRNA expression level of Bcl-2, FGF1 and FGF2 increased(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-1β, IL-6 and TNF-α in peripheral blood decreased (P<0.05). Conclusion: IFI16 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 Th1/Th2 cells and inhibiting the activity of pro-inflammatory factors. |
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