Effect Of Bortezomib Combined With Etoposide On The Expression Of NF-κB P65 MRNA In Hut-78 Cell Line

Objiective:In order to provide a theoretical basis for the clinical use of CTCL by explore the antitumor effects of Bortezomib(BTZ) combined with Etoposide(VP16),with detecting the expression of NF-kB P65 mRNA and its correlation in cutaneous T-cell lymphoma(CTCL) Hut-78 cell line.Methods:Hut-78 cells were cultured through cell lines and treated different concentrations of 2,5,10,50, 100nom/L of BTZ and 42.5,170,340,680, 1360,1700 umol/L of VP16 single or combined with eachother for 24 and 48 hours. The expression of NF-κB P65 mRNA were detected by the SYBR Green I Reverse transcription-polymerase chain reaction(RT-PCR), and then analysis the correlation between them.Results:1、When the Hut-78 cells was treated with a certain concentration of BTZ (24h-48h), the expression of NF-κB P65 mRNA level were lower than the control group with the increase of the extension of time and concentration, P<0.05.2、When the Hut-78 cells was treated with a certain concentration of VP16 (24h-48h), With the increase of the extension of time and concentration, the expression of NF-κB P65 mRNA level first decreased and then increased. The expression level were lower than the control group when the concentration of VP16 was 42.5umol/L (P<0.05). The expression level increased significantly at 48h, when the concentration of VP16 was 1360umol/L and 1700umol/L, P<0.05。3、The mRNA expression level ofNF-KB P65 in Hut-78 cells treated with BTZ and VP16 with a certain concentration.(1) When the Hut-78 cells was treated with a certain concentration of BTZ and VP16 for 24h and 48h, the expression of NF-κB P65 mRNA level were lower than the control group with the increase of the extension of time and concentration, P<0.05.(2) When the Hut-78 cells was treated with a certain concentration of BTZ and VP16, as compared to 24 hours and 48 hours, the mRNA expression level of NF-κB P65 for 24 hours were lower then 48 hours in the group which concentration lower then the one had the concentration of BTZ for 50nom/L and VP16 for 1360umol/L. On the contrary, the mRNA expression level of NF-κB P65 for 24 hours were higher then 48 hours in the group which concentration higher then the one had the concentration of BTZ for 50nom/L and VP16 for 1360umol/L.(3) When the Hut-78 cells was treated with a certain concentration of BTZ collaborated with VP16 for 24h and 48h, as compared to single group of VP16, the mRNA expression level of NF-κB P65 is on a downward trend with the concentration of combination group increased and lower then the single one.(4) When the Hut-78 cells was treated with a certain concentration of BTZ collaborated with VP16 for 24h and 48h, as compared to single group of BTZ, the mRNA expression level of NF-kB P65 is on a downward trend with the concentration of combination group increased, the level on a downward trend for single group of BTZ for 24 hours, On the contrary, the level higher then single group of BTZ for 48 hours.Conclusions:1. The mRNA expression level of NF-kB P65 could be down-regulated when treated with a certain concentration of BTZ only for 2nmol/L in Hut-78 cells.2. The mRNA expression level of NF-kB P65 could be first decreased when treated with a certain concentration of VP16 for 42.5umol/L,and the expression have a upward tendency with the concentration of VP16 increased in Hut-78 cells. The mRNA expression level of NF-kB P65 could be activated becase of the higher concentration and the level could increased with extension of time.3. The mRNA expression level of NF-kB P65 could be regulated by the collaboration of BTZ and VP16 when treated with appropriate concentration and time,the higher collaboration of BTZ could strongly balance the expression which activated by VP16.