Identification and Functional Characterization of a Novel Immunomodulatory Protein From Morchella conica SH.
Identification and Functional Characterization of a Novel Immunomodulatory Protein From Morchella conica SH.
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DOI：
10.3389/fimmu.2020.559770
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发表时间：
2020
影响因子：
7.3
通讯作者：
Tang X
中科院分区：
文献类型：
--
作者：
Wu G;Sun Y;Deng T;Song L;Li P;Zeng H;Tang X
在珍贵的药食两用蘑菇羊肚菌SH中发现了一种新型真菌免疫调节蛋白（FIP），命名为FIP-mco，属于FIP家族。使用邻接法对不同来源的 FIP 进行系统发育分析。研究发现，FIP-mco属于FIP家族的一个新分支，与大多数其他FIP相比，可能是从不同的祖先进化而来的。 FIP-mco的cDNA序列被克隆并在酵母毕赤酵母X33中表达。 FIP-mco重组蛋白(rFIP-mco)经琼脂糖Ni层析纯化，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western blot分析进行测定。 rFIP-mco蛋白在体外浓度分别为15和5μg/ml时可显着抑制A549和HepG2细胞的增殖，并分别在15和30μg/ml浓度下抑制人A549和HepG2细胞的迁移和侵袭。此外，rFIP-mco可以显着降低THP1细胞（人骨髓性白血病单核细胞）中TNF-α、IL-1β和IL-6的表达水平。为了探讨rFIP-mco对A549和HepG2细胞的细胞毒作用的潜在机制，对这两种癌细胞进行了细胞周期和凋亡测定。结果表明，G0/G1 期阻滞和细胞凋亡增加可能有助于 rFIP-mco 在两种癌细胞中抑制增殖。还通过抑制 NF-κB 信号通路研究了 rFIP-mco 减少炎症细胞因子作用的分子机制。结果表明，rFIP-mco 抑制 NF-κB 信号传导导致炎症细胞因子减少。这些结果表明来自 M. conica SH 的 FIP-mco 作为癌症治疗研究和医学应用的有效且可行的来源的前景。
A novel fungal immunomodulatory protein (FIP) was found in the precious medical and edible mushroom Morchella conica SH, defined as FIP-mco, which belongs to the FIP family. Phylogenetic analyses of FIPs from different origins were performed using Neighbor-Joining method. It was found that FIP-mco belonged to a new branch of the FIP family and may evolved from a different ancestor compared with most other FIPs. The cDNA sequence of FIP-mco was cloned and expressed in the yeast Pichia Pastoris X33. The recombinant protein of FIP-mco (rFIP-mco) was purified by agarose Ni chromatography and determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The protein rFIP-mco could significantly suppress the proliferation of A549 and HepG2 cells at the concentration of 15 and 5 μg/ml, respectively, and inhibited the migration and invasion of human A549 and HepG2 cells at the concentration of 15 and 30 μg/ml respectively in vitro. Further, rFIP-mco can significantly reduce the expression levels of TNF-α, IL-1β, and IL-6 in the THP1 cells (human myeloid leukemia mononuclear cells). In order to explore the potential mechanism of the cytotoxicity effect of rFIP-mco on A549 and HepG2 cells, cell cycle and apoptosis assay in the two cancer cells were conducted. The results demonstrated that G0/G1 to S-phase arrest and increased apoptosis may contribute to the proliferation inhibition by rFIP-mco in the two cancer cells. Molecular mechanism of rFIP-mco’s reduction effect on the inflammatory cytokines was also studied by suppression of the NF-κB signaling pathway. It showed that suppression of NF-κB signaling is responsible for the reduction of inflammatory cytokines by rFIP-mco. The results indicated the prospect of FIP-mco from M. conica SH as an effective and feasible source for cancer therapeutic studies and medical applications.
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