On The Studies Of Anti-tumor And Anti-virus Effect Of Marine Actinomycin X2

OBJECTIVEStudy on the biologic activity of marine actinomycin X2.The main research is about anti-tumor and anti-viral.METHODS1.The inhibition of marine actinomyein X2 on normal and tumor cells proliferation was detected by MTT method.Flow cytometer was used to detect the change of HepG-2 cell cycle,apoptosis and those disposed cells were stained with Hoechst 33258(8g/L).2.The inhibitory action of virus by use of different concentration marine actinomycin X2 by use MTT methods and study the effective of viral adsorption.Cytoplasmic RNA from ECV304 was extracted by the Trizol method and mRNA levels of ICAM-1, VCAM-1 were assayed by reverse transcript-polymerase chain reaction(RT-PCR) method respectively.Flow cytometer was used to detect the expression of ICAM-1, VCAM-1 on the infection ECV304 in different time courses respectively.Fluctuation of NF-κB was detected by semi-quantitative immunohistochemistry.RESULTS1.Marine actinomycin X2 has the inhibitory action to the tumor cells.FCM showed HepG-2 in G1 phase decreased,cells in S and G2/M phase increased,the cell cycle was arrested in S and G2/M phase.HepG-2 cell apoptosis with nuclear chromatin concentration and fragmentation as well as the formation of apoptotic bodies were observed.2.Marine actinomycin X2 can inhibit virus before absorption of CVB3,H1N1 into the normal cells and SI weree 63.88,47.66.3.The infection of CVB3 did promote the transcription of ICAM-1 and VCAM-1 in all 54h. The level of ICAM-1 and VCAM-1 mRNA in CVB3 group and CVB3+X2 group were higher than that of control group,and the differences had statistical significance(P＜0.05 ).The expression was increased in all 54h among which the VCAM-1 mRNA reached a peak at 12h point post-infection.There had expression of ICAM-1 mRNA in normal condition and increased after infection,especially between 24h-54h.In CVB3+X2 group at 12h-24h,12h-54h the expression of VCAM-1 and ICAM-1 mRNA was lower than CVB3 group(P＜0.05).4.At the same time,we observed the significance increased expression VCAM-1 and ICAM-1 protein on the surface of ECV304 by the way of FCM at 12h-54h.In CVB3+X2 group at 12h-24h,12h-54h the expression of VCAM-1 and ICAM-1 protein was lower than CVB3 group(P＜0.05).5.Our findings suggest that the level of NF-κB expression significantly increased in the CVB3 group aRer 3h.In CVB3+X2 group,at 1h,3h the level of NF-κB expression was increased,however,at 6h-24h the activation of NF-κB inhibited by X2.CONCLUSION1.Marine actinomycin X2 had significant inhibitory effect on tumor with dose and time-effect relationship.It arrested cell cycle,induced HepG-2 cells apoptosis and formed apoptosis bodies.4.The mechanism of anti-virus of Marine actinomycin X2 possibly arrested the absorption of virus.3.Marine actinomycin X2 decreased the expression of VCAM-1,ICAM-1 mRNA and protein maybe through inhibiting the activation of NF-κB.