Effects Of TRAIL Combined With Chenotherapeutics On Apoptosis Of HEP-2 And HNE-1 Cell Lines

Background:Chemotherapy is one of the most important treatments of nasopharyngeal carcinoma and laryngocarcinoma at present.Large dose of chemotherapeutics can kill not only tumor cells but also nomal cell and greatly reduce the quality of life of cancer patients.Therefore,finding a efficient way that can kill tumor cells and reduce the toxicity is a hot spot.It has been reported that tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) can kill the tumor cells,with minor side effects and low toxicity.It is expected to become a new antitumor drug.However,there are little reportes about the effects of nasopharyngeal carcinoma cell line (HNE-1) and human epidermal carcinoma cell line (HEP-2).This paper aims to study the joint effect of TRAIL+5-FU and TRAIL+DDP respectively on the proliferation and apoptosis of HEP-2,HNE-1 cell lines,and to explore its possible mechanism.Objective:To obsere the effects of different concentrations of TRAIL and chemotherapeutic drugs(5-FU,DDP)on HNE-1, HEP-2 cell lines proliferation,apoptosis ,both individually and jointly.Fourther more to explore the its possible mechanism.Methods:1 Different concentrations of TRAIL(1.0,10.0,100.0,1000.0ng/ml); DDP(1.0,10.0,10.0μg/ml) and 5-FU(1.0,10.0,100.0μg/ml) was used to test the inhibition of proliferation on HEP-2,HNE-1 cell lines by MTT.2 The experiment were divided into five groups:control group, TRAIL(10.0ng/ml) treatment group,DDP(1.0μg/ml) treatment group, 5-FU(1.0μg/ml) treatment group,DDP(1.0μg/ml)+TRAIL(10.0ng/ml) treatment group and 5-FU(1.0μg/ml)+TRAIL(10.0ng/ml) treatment groups. MTT assay was used to observe the inhibition of proliferation on HNE-1,HEP-2 cell lines in different treatment groups.3 Apoptosis rate were detected by FCM in different treatment groups. Cell morphology were observed by fluorescence microscopy.4 The expression of Celluar Fas-ssociated death domain-like IL-1 beta converting enzyme inhibitory protein(c-FLIP) and nuclear factorsкB (NF-кB) in the cell of different treatment groups was detected by Western-blot.Results:1 the research of TRAIL combined with 5-FU, DDP respectively on the HEP-2 cell line:1) Concentration of 1.0μg/ml of 5-FU and DDP can inhibit the proliferation of HEP-2 cell line respectively,concentration of 1.0ng/ml of TRAIL cannot inhibit the proliferation of HEP-2 cell line,but concentration of 10.0ng/ml and above of TRAIL can inhibit the proliferation of HEP-2 cell line.The concentration of 5-FU1.0μg/ml,DDP1.0μg/ml,TRAIL10.0ng/ ml was selected for further experiments.2) The inhibition rate of DDP+TRAIL-treated group(33.69±3.50)% was statistically significantly increased,comparing with TRAIL-treated group(5.73±4.31)% and DDP-treated group (15.06±8.83)%.The inhibition rate of 5-FU+TRAIL-treated(29.21±9.80)% was statistically significantly increased,comparing with TRAIL-treated group(5.73±4.31)% and 5-FUtre- ated group (12.68±5.20)%.3) The apoptosis rate of DDP+TRAIL-treated group(24.76±1.20)% was statistically remarkable improved,comparing with TRAIL-treated group(0.24±0.023)% and DDP-treated group(14.70±2.21)%.The apoptosis rate of 5-FU+TRAIL-treated group(14.33±0.78)% was statistically remarkable improved,comparing with TRAIL-treated group(0.24±0.023)% and 5-FU-treated group(2.25±0.56)%.4) The expression of c-FLIP,NF-кB of TRAIL-treated group, 5-FU-treated group and DDP-treated group was higher than that of TRAIL+5-FU-treated group and TRAIL+DDP-treated group.2 the research of TRAIL combined with 5-FU, DDP respectively on the HNE-1 cell line 1) Concentration of 1.0μg/ml of 5-FU and DDP can inhibit the proliferation of HNE-1 cell line respectively,concentration of 1.0ng/ml of TRAIL cannot inhibit the proliferation of HNE-1 cell line, and concentrati- on of 10.0ng/ml and above of TRAIL can inhibit the proliferation of HNE-1 cell line.The concentration of 5-FU 1.0μg/ml,DDP1.0μg/ml,TRAIL 10.0ng/ml was selected for further experiments.2) The inhibition rate of DDP+TRAIL-treated group(38.61±7.56)% was statistically significantly increased,comparing with TRAIL-treated group(5.73±4.42)% and DDP-treated group (14.62±7.92)%.The inhibition rate of 5-FU+TRAIL-treated(31.89±5.61)% was statistically significantly increased,comparing with TRAIL-treated group(5.73±4.42)% and 5-FUtre- ated group (14.17±9.6)%.3) The apoptosis rate of DDP+TRAIL-treated group (14.72±1.20)% was statistically remarkable improved,comparing with TRAIL-treated group(0.66±0.23)% and DDP-treated group (2.19±0.52)%.The apoptosis rate of 5-FU+TRAIL-treated group(14.70±1.78) % was statistically remarkable improved,comparing with TRAIL-treated group(0.66±0.23)% and 5-FU-treated group(2.43±0.26)%.4) The expression of c-FLIP,NF-кB of TRAIL-treated group, 5-FU-treated group and DDP-treated group was higher than that of TRAIL+5-FU-treated group and TRAIL+DDP-treated group.CONCLUSIONS: 1 The inhibition of proliferation and apoptosis of HEP-2,HNE-1 cell lines can be induced by TRAIL, and the effects were higher by TRAIL combined with 5-FU and DDP respectively;2 TRAIL can induce the tumor cells apoptosis by reducing the expression of c-FLIP,NF-кB in the process of apoptosis of HEP-2 and HNE-1 cell lines.