The Effects Of 2,2,6,6-Tetramethyl-3-Ene-Piperidine Nitroxide Rotenone Proxetil Anti-tumor Activity In Vitro

Objective:Tumor cells are usually out of control from normal growth regulatory system, they have some special physiological characteristics, including sustaining division and proliferation, losing of fidelity of cell cycle processes, migration ability and immortal. Almost all the anti-cancer researches are focused on the inhibition of cell proliferation, cell migration, cell cycle progression, and inducing cell apoptosis. PNR is a semi-synthesized compound with rotenone as substrate, and rotenone derived from plant extracts which has anti-tumor activity. The preliminary research of the effect of PNR on the tumor cell growth inhibitory, cell migration inhibitory, cell cycle arrest and cell apoptosis are performed in this thesis.Methods:Human tongue squamous cell carcinoma cell line Tca8113, human lung adenocarcinoma epithelial cell line A549, human hepatocellular liver carcinoma cell line HepG2, human gastric cancer cell line BCG823 and human urinary bladder cancer cell line EJ were stimulated by different concentration of PNR, respectively. The number of viable cells was estimated by the SRB assay. The motility of Tca8113 cells with or without PNR was evaluated by Transwell migration assay. Cell cycle of Tca8113 cells with or without PNR was tested by FCM. The apoptosis morphology of Tca8113 cells with or without PNR was observed by DAPI staining method.Results:PNR could inhibit the prolifration of Tca8113, A549, HepG2, BCG823 and EJ cells in vitro, and the effects were in a time-and dose-dependent manner on A549, HepG2 and BCG823 cells as the concentration of PNR from 0.8 to 12.8μg/mL. But, Tca8113 and EJ were only in a dose-dependent manner as the concentration of PNR from 2 to 7μg/mL. The sensitivity of PNR shows difference among these five tumor cells, Tca8113 cells was more sensitive than others, and the inhibition rate was 90% treated with PNR in a dose of 7μg·mL-1 for 48 hours. PNR could obviously inhibited Tca8113 cells migration, when the concentration of PNR is 6μg·L-1, the migration inhibitory rate was 72%.Cell cycle analysis showed accumulation of Tca8113 cells in the S phase after 24 and 48 hours of PNR treatment. Moreover, PNR could efficiently induce the apoptosis of Tca8113 cells, and the apoptosis rate reached 65.7% with PNR in a dose of 6μg·mL-1 after 24 hours of treatment.Conclusion:PNR has anti-tumor activity, it could inhibit the proliferation of tumor cells, especially against Tca8113 cells, and the mechanism is possibly by inducing the S phase cell cycle arrest and apoptosis.In addation, PNR could inhibit Tca8113 cell migration.