F K Guo1, S G Wu. 1. Institute of Pharmaceutic Sciences, First Military Medical University, Guangzhou 510515, China. jydyys@fimmu.edu.cn
Abstract
AIM: To investigate whether phosphatidylinositol (PI) 3-kinase is involved in interleukin-18 (IL-18)-induced nuclear factor-kappa B (NF-kappa B) activation. METHODS: Antisense PI 3-kinase oligonucleotide (ODN) was delivered by lipofectin encapsulation into cultured human hepatocellular carcinoma HepG2 cells. PI 3-kinase mRNA expression was assayed by semiquantitative reverse transcription-PCR. The levels of NF-kappa B were measured by sandwich ELISA. RESULTS: (1) Antisense PI 3-kinase ODN blocked PI 3-kinase mRNA expression. (2) IL-18 activated NF-kappa B from basal level of 0.153 +/- 0.008 to 1.942 +/- 0.017. (3) Antisense PI 3-kinase ODN inhibited IL-18-induced NF-kappa B activation in a concentration (1-8 mg/L)- and time (5-24 h)-dependent fashion. When the cells were treated with antisense PI 3-kinase ODN 2 mg/L for 8 h, a maximum inhibitory rate was 35.2% from 1.942 +/- 0.017 control to 1.259 +/- 0.018. CONCLUSION: PI 3-kinase is necessary for IL-18-stimulated NF-kappa B activation.
AIM: To investigate whether phosphatidylinositol (PI) 3-kinase is involved in interleukin-18 (IL-18)-induced nuclear factor-kappa B (NF-kappa B) activation. METHODS: Antisense PI 3-kinase oligonucleotide (ODN) was delivered by lipofectin encapsulation into cultured humanhepatocellular carcinoma HepG2 cells. PI 3-kinase mRNA expression was assayed by semiquantitative reverse transcription-PCR. The levels of NF-kappa B were measured by sandwich ELISA. RESULTS: (1) Antisense PI 3-kinase ODN blocked PI 3-kinase mRNA expression. (2) IL-18 activated NF-kappa B from basal level of 0.153 +/- 0.008 to 1.942 +/- 0.017. (3) Antisense PI 3-kinase ODN inhibited IL-18-induced NF-kappa B activation in a concentration (1-8 mg/L)- and time (5-24 h)-dependent fashion. When the cells were treated with antisense PI 3-kinase ODN 2 mg/L for 8 h, a maximum inhibitory rate was 35.2% from 1.942 +/- 0.017 control to 1.259 +/- 0.018. CONCLUSION:PI 3-kinase is necessary for IL-18-stimulated NF-kappa B activation.