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胃癌组织中存在较高的猪鼻支原体感染率,提示猪鼻支原体感染同胃癌的发生可能存在某种相关性.由于肿瘤坏死因子(TNF-α)在微生物感染导致肿瘤发生中起重要作用,而P37蛋白是猪鼻支原体的主要免疫原,因此,探讨P37能否诱导TNF-α的表达与释放,对阐明猪鼻支原体在胃癌发生中的可能分子机制有重要意义,运用PCR技术克隆了p37基因,在对其编码序列中7个色氨酸密码子TGA进行TGG定点突变后,利用PGEX-4T-1表达载体在大肠杆菌中成功表达了P37蛋白,并经Western blot得到证实,在此基础上,利用RT-PCR及TNF-α敏感细胞株L929的细胞毒性实验,发现P37确能诱导外周血单核细胞表达和释放TNF-α,抗P37单克隆抗体PD4对该诱导活性有明显的中和作用,结果提示,猪鼻支原体通过P37诱导TNF-α产生在猪鼻支原体感染所致相关疾病中可能起重要作用,值得进一步研究.
There is a high prevalence of Mycoplasma hyopneumoniae in gastric cancer tissues, suggesting that there may be some correlation between Mycoplasma hyorhinis infection and gastric carcinogenesis.As the tumor necrosis factor (TNF-α) plays an important role in tumorigenesis caused by microbial infection, P37 protein Is the main immunogen of Mycoplasma hyorhinis, therefore, to explore the P37 can induce the expression and release of TNF-α, to clarify the possible molecular mechanism of Mycoplasma hyorhinis in gastric carcinogenesis, the use of PCR cloned p37 gene, After the tryptophan codon TGA of seven tryptophan codons was mutated in TGG site, the P37 protein was successfully expressed in E. coli using PGEX-4T-1 expression vector and confirmed by Western blot. On the basis of this, PCR and TNF-α-sensitive cell line L929 cytotoxicity experiments and found that P37 can indeed induce the expression of peripheral blood mononuclear cells and the release of TNF-α, anti-P37 monoclonal antibody PD4 on the induction of significant neutralization effect, the results suggest , Mycoplasma hyorhinis can induce TNF-α production through P37 may play an important role in the related diseases caused by Mycoplasma hyorhinis infection, which deserves further study.