An Experimental Study Of The Effect And Associated Mechanisms Of Rh-Apo2L Combined With Vinorelbine On Lung Squamous Cell Carcinoma Cells

Objective：To investigate the effect of rh-Apo2L combined with vinorelbine on proliferationof lung squamous cell carcinoma cells（SK-MES-1）, and to probe into its possiblemechanism.Methods：1．To probe into the effect of rh-Apo2L or vinorelbine respectively on proliferationof lung squamous cell carcinoma cells（SK-MES-1）: SK-MES-1cells were treatedwith various concentration of rh-Apo2L or Vinorelbine for24~48hours. Proliferationinhibition rate of SK-MES-1cells were measured by CCK-8assay, and half inhibitoryconcentration (IC50) were analyzed.2．To probe into the effect of different groups on inducing apoptosis of lungsquamous cell carcinoma cells （SK-MES-1）: Divided the lung cancer cellsSK-MES-1into four groups: control, rh-Apo2L, vinorelbine, rh-Apo2L withvinorelbine, every group was treated with associated conditions and administrationsfor24hours. Morphology changes of SK-MES-1cells were observed throughmicroscopy. The apoptosis rates of different groups were detected by Flow cytometry.3．To probe into the expression of apoptosis-related protein in SK-MES-1cellsintervented by different group drugs: every group was treated with associatedconditions and administrations for24hours, Western blot was used to quantify thechange of the apoptosis-related protein DR4、DR5、caspase-3.Results：1．Rh-Apo2L could induce apoptosis in the lung squamous cell carcinoma cell lineSK-MES-1, presenting obvious dose-dependent cytotoxicity through CCK-8, and IC50of24h was28.57μg/ml, while IC50of48h was8.97μg/ml. vinorelbine also could induce apoptosis in the cell SK-MES-1, presenting obvious dose-dependentcytotoxicity through CCK-8, and IC50of24h was27.30μg/ml, while IC50of48h was7.87μ g/ml. The apoptosis rate of the lung squamous cell carcinoma cellline(SK-MES-1)was a significant difference to the group of rh-Apo2L (P<0.05).And synergistic effect of rh-Apo2L combined with vinorelbine was found, the druginteraction index (q value) were1.937(24h) and2.400(48h).2．The apoptosis rate of the lung squamous cell carcinoma cell line SK-MES-1wasobviously increased by intervention of rh-Apo2L combined with vinorelbine(25.64%), compared to the group of rh-Apo2L (16.80%) or vinorelbine（5.93%）treatment with a significant difference(P<0.05)according to the flow cytometry.3．The expression level of the apoptosis-related factors DR4, DR5and caspase-3was upregulated in group of rh-Apo2L combined with vinorelbine, and there was asignificant difference between different groups (P<0.05).Conclusions：1．rh-Apo2L and vinorelbine can inhibit the proliferation of SK-MES-1cells byinducing the cell apoptosis.2．rh-Apo2L combined with vinorelbine can apparently increase the apoptosis rateof the lung squamous cell carcinoma cell line(SK-MES-1).3. The expression level of the apoptosis-related factors DR4, DR5and Caspase-3may be involved in the mechanism increased apoptosis.