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【目的】探讨Hp cagⅡ对胃上皮细胞IL-8基因转录的影响及信号传导机制。【方法】构建cagⅡ基因位点缺失Hp突变株及带有IL-8报告基因的人胃癌细胞系L5F11,用液体闪烁计算仪测定荧光素酶(IL-8转录)活性,用ELISA法测定IL-8蛋白浓度。【结果】所有Hp突变株诱导荧光素酶活性与IL-8蛋白浓度较亲代菌株26695均降低(0.13±0.01vs0.59±0.05×10~6cmp,P<0.01;0.73±0.13vs2.22±0.65ng/ml,P<0.05)。PTK抑制剂herbimycin A不仅抑制Hp诱导的荧光索酶活性(0.71±0.18vs1.51±0.23×10~6cpm,P<0.05),而且抑制IL-8蛋白表达(0.83±0.41vs3.22±0.59ng/ml,P<0.05),但herbimycin A对TNF_α诱导的荧光素酶活性及IL-8蛋白表达均无影响(P均>0.05);PKA抑制剂H7抑制TNFa诱导的荧光素酶活性(0.74±0.16vs2.62±0.26×10~6cpm,P<0.001)及IL-8蛋白表达(1.45±0.38vs4.12±0.43ng/ml,P<0.01),而对Hp诱导的荧光素酶活性无影响(P>0.05)。【结论】Hp cagⅡ中的多个基因能够调节胃上皮细胞IL-8基因转录,且这一作用主要经蛋白酪氨酸激酶途径。
【Objective】 To investigate the effect of Hp cagⅡ on the transcription of IL-8 gene in gastric epithelial cells and the signal transduction mechanism. 【Methods】 Human gastric cancer cell line L5F11 with deletion of cagⅡ locus and IL-8 reporter gene was constructed. Luciferase (IL-8 transcription) activity was measured by liquid scintillation counting instrument. IL- 8 protein concentration. 【Result】 All of the Hp mutants showed lower luciferase activity and IL-8 protein concentration than the parent strain 26695 (0.13 ± 0.01 vs0.59 ± 0.05 × 10 ~ 6cmp, P <0.01; 0.73 ± 0.13 vs2.22 ± 0.65 ng / ml, P <0.05). The herbicide PTK inhibitor herbimycin A not only inhibited the Hp-induced fluorescence of cordycepin (0.71 ± 0.18vs1.51 ± 0.23 × 10 ~ 6cpm, P <0.05), but also inhibited the expression of IL-8 protein (0.83 ± 0.41vs3.22 ± 0.59ng / ml, P <0.05). However, herbimycin A had no effect on TNF-αinduced luciferase activity and IL-8 protein expression (all P> 0.05); PKA inhibitor H7 inhibited TNFa-induced luciferase activity 0.16vs2.62 ± 0.26 × 10 ~ 6cpm, P <0.001) and IL-8 protein expression (1.45 ± 0.38vs4.12 ± 0.43ng / ml, P <0.01), but had no effect on Hp-induced luciferase activity (P> 0.05). 【Conclusion】 Multiple genes in Hp cagⅡ can regulate the transcription of IL-8 gene in gastric epithelial cells, and this effect is mainly through the protein tyrosine kinase pathway.