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【目的】研究支原体对体外培养细胞基因功能的影响。【方法】利用si RNA抑制DNA双链损伤修复关键基因——FIGNL1在无支原体感染、感染猪鼻支原体及清除支原体污染后的人小细胞肺癌细胞株NCI-H446与NCI-H1688中的表达后,采用定量PCR、流式细胞术等方法检测支原体感染对宿主细胞目标基因表达、细胞周期等影响。【结果】虽然猪鼻支原体感染对si RNA沉默FIGNL1表达无显著影响,无支原体感染及清除支原体污染的H1688和H446细胞FIGNL1表达沉默后,与仅加转染试剂的空白组(mock)相比较,靶标FIGNL1基因的实验组(T1)和阴性对照组(nc)的S期细胞比例均未发生显著变化。但猪鼻支原体感染的H1688和H446细胞相对于空白组,实验组与阴性对照组的S期细胞比例,H1688细胞提高了约1.38倍和0.51倍,H446细胞提高了约1.27倍和0.55倍。【结论】推测由于支原体会对宿主细胞DNA造成损伤,而FIGNL1是DNA双链断裂损伤修复的重要基因,从而导致沉默猪鼻支原体感染的H1688和H446细胞FIGNL1表达时,细胞会出现S期阻滞。鉴于支原体感染对细胞有广泛、显著的影响,在基因功能、肿瘤等细胞生物学研究中,应予以高度重视。
【Objective】 To study the effect of mycoplasma on the gene function of cultured cells in vitro. 【Method】 RNAi was used to inhibit the expression of the key gene ofIGNFL1 in NCI-H446 and NCI-H1688 cells after mycoplasma infection without Mycoplasma infection, Mycoplasma hyorhinis and Mycoplasma pneumoniae. Quantitative PCR, flow cytometry and other methods to detect mycoplasma infection host cell target gene expression, cell cycle and so on. 【Results】 Although Mycoplasma hyorhinis infection did not affect the expression of FIGNL1, the expression of FIGNL1 in H1688 and H446 cells without mycoplasma infection and Mycoplasma removal was significantly lower than that in blank group (mock) There was no significant change in the percentage of S phase cells between the experimental group (T1) of the FIGNL1 gene and the negative control group (nc). However, the proportion of S phase cells in H1688 and H446 cells infected with Mycoplasma hyopneumoniae was significantly increased by 1.38-fold and 0.51-fold compared with that of blank control group and H1688 cells respectively, and 1.27-fold and 0.55-fold increase of H446 cells. 【Conclusion】 It is speculated that Mycoplasma can damage DNA of host cells, while FIGNL1 is an important gene for repair of DNA double-strand breaks, leading to S phase arrest when FIGNL1 is silenced in H1688 and H446 cells. In view of mycoplasma infection on the cells have a broad and significant impact in gene function, cancer and other cell biology research, should be highly valued.