Expression of Beclin1 and HLA Ⅰ , Ⅱ in human SKOV3 ovarian cancer cells and their correlation / 临床与实验病理学杂志

ABSTRACT

Purpose The aim of this study was to investigate the relationship between autophagy gene Beclin1 and immune response effector classical HLA Ⅰ,Ⅱ in SKOV3 cells.To explore the role of Beclin1 in immunity in ovarian cancer cells which were transfected with the vector of Beclin1.Methods RT-PCR and Western blot were used to detect the expression of Beclin1 and HLA Ⅰ,Ⅱ in SKOV3 cells.Fluorescence microscope was carried out to observe the unique autophagosome in SKOV3 cells.MTT was used to analyze the proliferation of the Beclin1 over-expressed SKOV3 cells.Results Transfection SKOV3 cells with Beclin1 vector could induce Beclin1 transcription and translation approximately 5 and 2 times compared with empty vector group respectively.The autophagosome stained by MDC was observed by fluorescence microscope.And much more green fluorescence signal was observed in Beclin1 vector group.RT-PCR and Western blot indicated that HLA Ⅰ,Ⅱ induced by transfection with extrinsic Beclin1.The allelic transcriptions of HLA Ⅰ-A,B,C and HLA Ⅱ-DP,DQ,DR in extrinsic Beclin1 group were approximately 2,1.6,3 and 2,6,3 times compared with empty vetcor group or untreated group,respectively.The results of Western blot showed that HLA Ⅰ,Ⅱ in Beclin1 vector group induced as much as 2 and 1.6 times compared with empty vetcor group or untreated group,respectively.The results of MTT showed that the proliferation of SKOV3 cells treated with Beclin1 vector was significantly suppressed.The percentage of suppression in Beclin1 vector group at 24 h,48 h,72 h and 96 his42.6％,37.8％,24.35％,14.81％ compared with untreated group or empty vector group respectively.Conclusion The enhancement of autophagy by over-expression of Beclin1 could induce HLA Ⅰ,Ⅱ transcription and translation in SKOV3 cells.The expression of HLA Ⅰ,Ⅱ may be responsible for triggering the immune response in ovarian cancer.Over-expression of Beclin1 could inhibit the proliferation of SKOV3 cells which were transfected with extrinsic Beclin1.