Signaling Pathways Involved In Expression Of Genes Induced By Cyadox

Cyadox, a new derivative of quinoxalines, has been ascertained as an antibiotic with significant growth promoting, low poison, quickly absorption, swift elimination, brief residual period and non-cumulative effect.7differential expressed genes, include Insulin-like Growth Factor-1, Epidermal Growth Factor, Poly ADP-ribose polymeras, The Defender Against Apoptotic Death1, Complement Component3, Transketolase and a new gene, were induced by cyadox in swine liver tissues, which were found by mRNA differential display reverse transcription polymerase chain reaction (DDRT-PCR) in our laboratory. DAD1was identified as a negative regulator of programmed cell death. C3could enhance the immune system by activating the complement system. TK, a key point of pentose phosphate pathway, influenced many cells vital movement such as cell proliferation by regulating the metabolism of glucose. EGF and IGF-1belong to growth factor class, play an important role in cell growth. PARP is an immediate DNA-damage dependent post-translational modification of histones that contributes to the survival of injured proliferating cells. The antibiosis and growth promotion effect of cyadox may have be related with7genes. This research can help us to find out the signaling pathways which were involved in expression of7gens induced by cyadox.1.Signaling pathways involved in the expression of genes induced by cyadox in PK-15cellsThe mRNA expression levels of IGF-1, EGF, C3, DAD1, TK, PARP and New gene were detected by RT-qPCR after PK-15cells were exposed to cyadox (2μM) for0.5-8h. We found that cyadox treatment evoked a significant time-dependent effect of7genes. The mRNA expression levels of EGF, C3, DAD1, TK, PARP and New gene all increased at1h, the mRNA expression of IGF-1increased at2h. In the research of signaling pathways involved in expression of7genes induced by cyadox, the protein expressions and activations of P38, PAKT, P44/42, and P65were assessed by western blotting when PK-15cells were treated with cyadox (2μM) or signal pathway inhibitors for4h. The mRNA expression levels of IGF-1, EGF, C3, DAD1, TK, PARP and New gene were detected by qRT-PCR. These results suggested that the activation of IGF-1induced by cyadox might be P13K&NF-KB-dependent, Erk1/2&JNK may play assistance role. The activation of TK induced by cyadox might be NF-κB dependent, Erkl/2, JAK, PI3K, TGF-P may play assistance role. The PI3K, TGF-β&NF-κB play a significant role in mRNA expression of C3, Erkl/2may play assistance role. The activation of PARP might be P13K&NF-κB dependent, TGF-β&Erkl/2may play assistance role. The PI3K plays a significant role in expression of DAD1, however, NF-κB, JAK&TGF-P may play an assistance role in expression of DAD1. The mRNA expression level of EGF might be P13K&NF-κB dependent. The PI3、Erkl/2&NF-κB play a significance role in expression of New gene. These results suggest that the PI3K&NF-κB were the significant signal pathway involved in the expression of7genes induced by cyadox.2.Signaling pathways involved in the expression of genes induced by cyadox in primary cultured pig hepatocytesThe mRNA expressions of IGF-1, EGF, C3, DAD1, TK, PARP and New gene were detected by RT-qPCR after primary cultured pig hepatocytes were exposed to cyadox (2μM) for0.5-8h. We also found that cyadox treatment evoked a significant time-dependent effect of7genes. The mRNA expression levels of New gene, EGF and DAD1increased in0.5h; The mRNA expression levels of IGF-1, TK, PARP increased in1h, The mRNA expression level of C3increased in2h. In the signaling pathways involved in expression of7genes induced by cyadox research in primary cultured pig hepatocytes, The mRNA expression levels of IGF-1, EGF, C3, DAD1, TK, PARP and New gene were detected by RT-qPCR. These results suggested that the expression of EGF induced by cyadox might be P13K dependent. The expression of IGF-1induced by cyadox might be PI3K&JNK-dependent. The TGF-β play a significant role in mRNA expression of C3. The expression of DAD1might be TGF-β&P13K-dependent. The TGF-β, P38, PI3K play a significant role in mRNA expression of PARP. The expression of TK might be Erkl/2, P38, TGF-β, JNK, P13K-dependent. The P38, TGF-β, JNK, P13K play a significant role in mRNA expression of New gene. These results suggest that the PI3K were the significant signal pathway involved in the expression of7genes induced by cyadox in primary cultured pig hepatocytesThese researches showed that the mechanism of signaling pathways involved in7genes induced by cyadox were variegated between PK-15cell and primary cultured pig hepatocytes. The PI3K signaling pathway was the principal signaling pathway which could regulated the expression of7gene related to cyadox. However, the time-effect relationships and principal signaling pathway of7genes expression induced by cyadox are different. The NF-κB signaling pathway paly a vital role in PK-15cell other than primary cultured pig hepatocytes.