The Study Of The Expression MHC Class â… \â…¡ Antigens, B7-1\B7-2 Moleculars After Augmenter Of Liver Regeneration Expression Blocked In HepG2 Cell

Objective:The expression of hALR in HCC (hepatocellular carcinoma) was observed very high with immunocytochemistry.And the proliferation of rat spleen mononuclear cells activated with HepG2 membrane antigens were inhibited by ALR.ALR was concluded which played an important role not only in escaping of tumor cells from immune surveillance but also in HCC auto-proliferation as hepatotrophic factors and immune regulators. The objective of the study is to observe in nucleic acid and protein levels the difference of MHCⅠ,MHCⅡ,CD80,CD86 expression in HepG2 cells after transfection SiRNA-ALR(augmenter of liver regeneration) . To investigate the ALR involved immune regulation in HCC has relationship with MHCⅠ,Ⅱ,CD80,CD86 or not .This result will provide the experimental evidence for further research the role of ALR in HCC immune escaping from immune surveillance.Methods:In this study,the RNAi plasmid PSiALR-A(by siRNA targeting hALR) and the unrelated control plasmid PSiALR-B were transfected into HepG2 cells respectively.At the same time,cultured HepG2 cells were devided into two groups for blank controls and lipofectamine reagents controls.The expression of MHCⅠ,Ⅱ,CD80,CD86 were detected by immunocytochemistry,RT-PCR,Western blot and FACS after transfection 48 h.All measurement data from RT-PCR,IHC,Western blot were expressed by mean±standard deviation,then analyzed by One-Way analysis of variance and q-test.P<0.05 express statistical significance,P<0.01 express predominant significance.All data analyzed by SPSS 12.0 statistical soft.Results : Immunocytochemical staining showed there were light positive expression of DR,CD80,CD86 moleculars and positive expression of MHCⅠantigens contrasted Raji cells by SP assay of immunocytochemistry.Although the expression of MHCⅠ,DR antigens in HepG2 blank controls group was significantly higher compared with lipofectamine reagents controls group (P<0.05),but importantly there is no difference between PSiALR-B/HepG2 group and PSiALR-A/HepG2 group(P>0.05).Expression of CD80,CD86 in PSiALR-B / HepG2 group and PSiALR-A /HepG2 group was no significantly difference(P>0.05).The RT-PCR showed expression of MHCⅠ,DR,CD80,CD86 in different groups include PSiALR-B/HepG2 group and PSiALR-A/HepG2 group was no visible changes after transfection.The results of Western blot show the same findings as the results of RT-PCR.The MHCⅠantigens in HepG2 cells detected by FACS among four groups were no inverse changes after transfection.There is no difference between PSiALR-B/HepG2 group and PSiALR-A/HepG2 group in the expression of DR,CD80,CD86 detected by FACS.But after transfection,expression of DR,CD80,CD86 were more higher than HepG2 blank control group or lipofectamine reagents control group.Conlusion:In nucleic acid and protein levels,the expression change of MHCⅠ,DR,CD80,CD86 in HepG2 cells was not observed after hALR expression blocked.It was suggested that hALR didn't play an imporant role in regulating surface moleculars expression of HepG2.