Study On Screening And Mechanism Of Indole Derivative 3IP

To investigate the screening and anti-tumor mechanism of 3-indoleethyl pentanamide (SIP) . 3IP was isolated and purificated by plant chemical methods, drug screening of fractions from Xenorhabdus nematophilus Bl . The in vitro anti-tumor activity of 3 IP obtained was tested using MTT assay. The in vivo anti-tumor activity of 3 IP was performed on nude mice inoculated HeLa cells in the back.Agarose gel electrophoresis of DNA and flow cytometry were used to detect the apoptosis of HL-60 cells and the activity of Caspase-8 and Caspase-3 were detected by Caspase Colorimetric Assay Kits.Indirect immunofluorescence assay was used to detect the expression of Bcl-2. Study on antitumor mechanism of 3IP used establishment of targeting API screening model which is based on transcription regulation of API (signal transducers and activators of transcription API).3-indoleethyl pentanamide (3IP) from Xenorhabdus nematophilus Bl was found;the IC50 of 3IP obtained for HepG2 , K562, HT-29 , BGC-823 and HeLa equal to 0.12 0.01 g/mL, 5.90 1.17 g/mL, 6.16 0.52ug/mL, 1.45 0.12 g/mL and 1.83 0.02 g/mL, respectively. No significant effects of 3 IP on the proliferation of normal human cells MRC-5 and L-02 were observed .3IP exhibited potential anti-tumor activity against HeLa with the percents of inhibition of tumor growth of 48.1 %, 56.5%, 72.2% at the doses of 2mg/kg, 4mg/kg, 8 mg/kg respectively. Apoptosis of HL-60 cells could be efficiently induced by 3IP. The activity of Caspase-3 of induced HL-60 cells was remarkably increased and the activity of Caspase-8 was not changed obviously after 3IP treatment. Bcl-2 gene expression was down-regulated with the concentration of 3 IP increasing. Also 3 IP can restrain the expression of the SEAP reporter gene .3IP was isolated and purificated by plant chemical methods from Xenorhabdus nematophilus B1,3IP has the potential anti-tumor activity in vitro and vivo,and it has little effect on the proliferation of normal human cells. Also 3IP can induce apoptosis of HL-60 cells,the apoptotic peaks were found by flow cytometry and "DNA ladder"appeared on 1.8% agarose gel. The mechanism of apoptosis have relations with up-regulation of Caspase-3 activity and down-regulation of Bcl-2 gene expression . The anti-tumor activity of 3 IP was contacted with AP1 signal transduction pathway by using screening model of targeting AP1.The results of our research provide experimental foundations for us to make further study on the mechnism of 3IP.