Gao Yifan, Padhiar Arshad Ahmed, Wang Jia, Zhang Wei, Zhong Mintao, Liu Ben, Kang Zhijie, Wang Xiaoli, Li Xingyun, Huang Min
Department of Microbiology, Dalian Medical University, Dalian, Liaoning, China; Department of Biology, Shenyang Normal University, Shenyang, Liaoning, China.
Department of Microbiology, Dalian Medical University, Dalian, Liaoning, China.
Gene. 2018 Feb 5;642:212-219. doi: 10.1016/j.gene.2017.10.080. Epub 2017 Oct 28.
Lentinula edodes C91-3 is an edible mushroom that has demonstrated a remarkable anti-tumor effect in various cancer cells both in vitro and in vivo. In the present study, we report the ability of recombinant thioredoxin-like latcripin 11 (LP-11) of Lentinula edodes C91-3 to suppress the proliferation of various cancer cells. The LP-11 gene of Lentinula edodes C91-3 was cloned in the pET-32a(+) expression vector and expressed in a prokaryotic system. The expressed protein was refolded by gradual dialysis and purified by affinity gel filtration chromatography. The antioxidant activity of LP-11 was tested by 1,1-dipheny l-2-picrylhydrazyl (DPPH) assay. The anti-tumor activity of recombinant LP-11 was tested in eight kinds of tumor cell lines by CCK-8 assay. Recombinant LP-11 significantly suppressed the proliferation of various cancer cells, but not normal human umbilical vein endothelial cells. Human lymphoma U937 cells exhibited the most sensitivity to LP-11 protein. U937 cell apoptosis was assessed by Annexin V staining coupled with flow cytometry, and mitochondrial morphology was analyzed by light and electron microscopy. It was revealed that recombinant LP-11 induced apoptosis in human leukemic monocyte lymphoma U937 cells. Our findings suggest that recombinant LP-11 is a promising agent for the treatment of lymphoma.
香菇C91-3是一种食用菌,已在体外和体内的各种癌细胞中显示出显著的抗肿瘤作用。在本研究中,我们报道了香菇C91-3的重组硫氧还蛋白样香菇素11(LP-11)抑制各种癌细胞增殖的能力。香菇C91-3的LP-11基因被克隆到pET-32a(+)表达载体中,并在原核系统中表达。表达的蛋白通过逐步透析复性,并通过亲和凝胶过滤色谱法纯化。LP-11的抗氧化活性通过1,1-二苯基-2-苦基肼(DPPH)测定法进行测试。重组LP-11的抗肿瘤活性通过CCK-8测定法在八种肿瘤细胞系中进行测试。重组LP-11显著抑制了各种癌细胞的增殖,但对正常人脐静脉内皮细胞没有影响。人淋巴瘤U937细胞对LP-11蛋白表现出最高的敏感性。通过膜联蛋白V染色结合流式细胞术评估U937细胞凋亡,并通过光学和电子显微镜分析线粒体形态。结果表明,重组LP-11诱导人白血病单核细胞淋巴瘤U937细胞凋亡。我们的研究结果表明,重组LP-11是一种有前途的淋巴瘤治疗药物。