Experimental Research Of Crizotinib And Tae684’s Effects On Eml4-Alk-Positive Cell Line H2228

Objective To study the effects of Crizotinib (PF-02341066) and TAE684(NVP-TAE84) on the cell’s proliferation and apoptosis of EML4-ALK-positive cell line H2228, with the purpose of providing the basis for clinical medication.Meterials and Methods EML4-ALK-positive human non-small lung cancer cell line H2228was kindly provided by Lung Cancer Institute of Guangdong People’s Hospital, cultured routinely to the logarithmic growth phase. To find out the half maximal inhibitory concentration of Crizotinib and TAE684. Set the drug gradients for our experiment. MTT assay was used to analysis the cells’ proliferation that separately treated by Crizotinb and TAE684for72h, respectively. Flow cytometry (FCM) was used to analysis the cells’apoptosis situation that treated by Crizotinib for24,48,72h, respectively. SPSS19.0statistical software was used to Process data. Cell’s growth rate was estimated by ANOVA analysis, p<0.05showed statistical difference. Regression and Probit analysis were used to estimate the drugs’half inhibitory concentration (IC50),95%confidence interval was chosen. Results Crizotinib induced the early apoptosis of H2228cells, inhibited the cell proliferation. In the concentration of10nM/L,30nM/L,90nM/L,270nM/L and810nM/L, the inhibition rates of Crizotinib on H2228cells were3.39%,10.51%,22.06%,29.60%,80.23%, respectively. The inhibition was increased by the increasing of the drug’s concentration (p<0.05). IC50for Crizotinib on H2228cells was334.51nM/L. In the concentration of335nM/L, the early apoptotic rates of the H2228cells, that dealt with Crizotinib for24h,48h,72h, were separately95.3%,90.0%and68.8%. The apoptosis rate decreased with the time extension (p<0.01). The necrosis rate of the H2228that untreated and treated by Crizotinib for24h,48h,72h, were separately0.1%,2.8%,2.0%,0.6%, respectively, showing no significant difference between the cells(p>0.05). TAE684showed no inhibition in the H2228cell’s proliferation.Conclusion Crizotinb has potent inhibition against EML4-ALK-positive cell line H2228. The inhibition increases by concentration-dependent manner. That is consistent with literature reports. IC50for Crizotinib on H2228cells is334.51nM/L. In the same concentration of Crizotinib, the apoptotic rate of H2228cells’ decreases with the time extension. The effect of Crizotinib on H2228cells occur on early apoptosis, with little effect on necrosis. TAE684shows no inhibition in the H2228cell’s proliferation in our experiment, which is consistent in part with previous reports.