Investigate The Drug-Resitance In Human Hepatoma Cell Line HEPG₂/Cddp/2.0by Interfere With Cell Cycle

Objective interferencing p15with shRNA, to investigate thedrug-resitance in human hepatoma cell line HepG₂/CDDP/2.0by interferewith cell cycle.Method Progressively increasing doses of CDDP was used inHepG₂, a human HCC cell line, to establish a CDDP-resistance cell line.Three specific shRNAs targeting p15gene （named Y1, Y2, and Y3） andone scrambled control shRNA were synthesized and individuallytransfected into HepG₂/CDDP/2.0cells using lipofectamine method.Screening best clip with RT-PCR.The cell viability were evaluated by3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT）.Cell cycle was assessed by flow cytometry. The influence ofshRNA-mediated p15deficiency on the expression of p15,p15、Bcl-2andBax was analysised by western blot.Results A CDDP-resistance cell line, HepG₂/CDDP/2.0, whichshowed moderate resistance to CDDP, was successfully established. TheY3shRNA had the highest gene-targeting efficacy. Remarkably reduced CDDP-induced apoptosis in HepG₂/CDDP/2.0.Cell proliferated quickly.Cell cycle reduced at G1. Knock down of p15significantly promoted theexpression of Bcl-2and deteriorated the expression of p15、Bax.Conclusion The percentage of cells at the G1phase increases with thelength of exposure to drugs in HepG₂/CDDP/2.0cells. Silencing p15cansignificantly decrease the fraction of G1subpopulation. Targeted reductionof p15decreases Bax expression and increases Bcl-2expression, whichmay consequently contribute to reduced apoptotic potentials and elevatedCDDP resistance in HepG₂/CDDP cells.