Effect Of SM-164Combined With Doxorubicin On Proliferation、Apoptosis And Cell Cycle In Human Osteosarcoma HOS Cell Line

Background and Objective：The new anti-cancer drug SM-164is a class of smallmolecule Smac compounds, this study aims to explore effect of the SM-164enhanced Doxorubicin on proliferation、induces apoptosis and cell cycle in humanosteosarcoma HOS cells, to further explore the mechanism of action and improvethe efficacy of clinical cancer chemotherapy provide an experimental basis.Method:1、MTT assay (1nmol/L,10nmol/L,30nmol/l,100nmol/l,200nmol/L,300nmol/L,1mmol/L, micromol/L,10micromol/L) SM-164,(0.25μg/mL,0.5μg/mL) Doxorubicin stars and their combination (24h,48h)inhibited the proliferation of human osteosarcoma HOS cells. Book of Q values toevaluate the combination effect.2、The Flow cytometry SM-164and Doxorubicin single-agent or combinationtherapy induced apoptosis of human osteosarcoma HOS cells. Flow cytometry todetect the SM-164and doxorubicin monotherapy or combination therapy onhuman osteosarcoma HOS cell cycle impact.Results：1、MTT results (1nmol/L,10nmol/L,30nmol/L,100nmol/l,200nmol/L,300nmol/L,1mmol/L, micromol/L,10micromol/L) of SM-164HOS cells after24h the survival rate was (86.63±6.58)%,(84.45±7.41)%,(82.76±7.12)%,(78.88±5.12)%,(76.97±4.26),(75.41±4.95)%,(72.99±3.41)%,(68.46±2.45)%,(63.63±1.45)%. SM-164Human Osteosarcoma HOScells the inhibitory effect of the drug was dose-dependent. Single Doxorubicin(0.25μ g/mL,0.5μg/mL),24-hour cell inhibition rate (21.41±0.14)%,(30.84±0.34)%,48-hour cell inhibition rate were (52.81±1.75)%,(63.88±2.73)%.SM-164200nmol/L combined with Doxorubicin Star (0.25μg/mL,0.5μg/mL), theinhibition rate of the24-hour HOS cells were (46.19±2.30)%,(53.49±1.06)%Q value1.65±0.09,1.46±0.05;48-hour HOS cells inhibition rate (73.86±1.59)%,(81.37±3.26)%, Q values were1.34±0.02,1.24±0.01. Combinationof Q values>1.15. The two-drug combination therapy inhibition rate wassignificantly higher, and the inhibitory effect of a dose-and time-dependent manner.2、Flow cytometry apoptosis of doxorubicin Star and SM-164can induce apoptosisin human osteosarcoma HOS cells, combination therapy induced apoptosis ratealong with it with the drug concentration increments. SM-164200nmol/Lmonotherapy role HOS cells24hours apoptosis rate was (14.46±3.86)%; thedoxorubicin (0.25μg/mL,0.5μg/mL) single-agent role HOS cells24hours apoptosisrate (26.67±3.67)%,(35.37±4.80)%, respectively; SM-164200nmol/Lcombination of doxorubicin Star (0.25μg/mL,0.5μg/mL) the role of HOS cellsapoptosis rates were24hours (32.26±1.14)%,(50.97±4.82)%. SM-164200nmol/L combined the doxorubicin0.25μg/mL role HOS cells when the Q valueis0.89, and its role as an additive effect; SM-164200nmol/L combination thedoxorubicin0.5μg/mL role HOS cells the Q value of1.16, and its inhibitory effectof the synergy. Flow cytometry cell cycle results show that doxorubicin-inducedosteosarcoma HOS cells apoptosis by inducing cell cycle arrest in the G2/M phase,Sm-164combination of doxorubicin on apoptosis in human osteosarcoma HOScells the effect may be interference in the growth and proliferation of cells in Sphase and arrested in S phase in advance.Conclusion：Smac-mimic SM-164and Doxorubicin has an explicit effect ofinhibiting proliferation of HOS cells in dose-dependent manner. SM-164couldinduce apoptosis and has a synergistic interaction with Doxorubicin, it possiblythrough the block at S phase enhance the effect of Doxorubicin on the proliferationinhibition of HOS cells.