Independent R&D Products-the Antitumor Activity Of Magnesium Cantharidate In Vitro And In Vivo And The Mechanism Research On Magnesium Cantharidate

Objective:Using Cantharidin acid magnesium which is self-developed carry anti-tumorexperiments in vivo that in order to provide early experimental works for the developmentof new drugs base on cantharidin acid magnesium.Methods:1, The proliferation inhibition of human hepatoma SMMC-7721,HepG2,BEL-7402cells was detected by used different concentrations of Cantharidin acidmagnesium, cantharidin, Cantharidin acid sodium by sulfonyl rhodamine staining method(SRB) in vitro;2,The cell morphological changes of different concentrations ofCantharidin acid magnesium, cantharidin, Cantharidin acid sodium on the three hepatomacells was observed by inverted phase contrast microscope;3,The cell cycles and apoptosisof Cantharidin acid magnesium on human hepatoma SMMC-7721was observed throughflow cytometry.4, The impact of Cantharidin acid magnesium on human hepatomaSMMC-7721was observed through HE staining and Hoechst33342staining.5, TEM wasused observed ultrastructural changes of Cantharidin acid magnesium on human hepatomaSMMC-7721.6, Establish human hepatoma SMMC-7721cells in nude mice xenograftmodel, the experimental group were randomly divided into saline, magnesium groupcantharidin cantharidin group, cantharidin sodium group, tumor volume measurement,weight;7, the impact of HE staining magnesium cantharidate on human hepatocellularcarcinoma xenograft tissue morphology;8, TEM magnesium cantharidate roledescendants HCC xenograft tissue ultrastructural changes.Results:1, magnesium cantharidin on SMMC-7721, HepG2, BEL-7402hepatoma cell linethree better than cantharidin and cantharidin, sodium cantharidate, and magnesiumcantharidate which cantharidin on human hepatoma SMMC-7721cell inhibition best;2,inverted phase contrast microscope: magnesium cantharidate、 cantharidin、sodiumcantharidate acting on the three hepatoma cells, from a low concentration group began thenumber of cells decreased gradually decreased with the increase in the number of cells indrug concentration, there are cell wrinkle shrink, smaller size, round and some celldisruption phenomena;3, flow cytometry results showed that: human hepatoma cellSMMC-7721in the role of magnesium cantharidate, G0/G1phase cells decreased, G2/ M phase cells increased cell appears G2/M phase arrest; cell apoptosis with magnesiumcantharidate concentration increased gradually increased.11.3%apoptosis rate increasedfrom0.8percent IC50concentration of the control group when;4, HE staining,Hoechst33342stained microscopic Show: human hepatoma cell SMMC-7721apoptoticcell morphology;5, TEM observation: the nucleus Alien chromatin clump edge set, seeapoptotic bodies;6, magnesium cantharidate tumor weight was significantly less than thesaline group (P <0.01), cantharidin group, sodium cantharidate tumor weight andphysical saline group, the difference was not statistically significant;7, HE stainingShow that magnesium cantharidate group transplanted tumor cells smaller size,cytoplasmic condensation, eosinophilic change, nuclear condensation, fragmentation.8, TEM visible magnesium cantharidate group tumor disappeared nuclear membrane,nuclear chromatin aggregation changed into a group and so on.Conclusion:1, magnesium cantharidate hepatoma cell line was better than cantharidin andcantharidin sodium;2, magnesium cantharidate can inhibit The proliferation and growthof tumor;3, magnesium cantharidate has effection on hepatoma SMMC-7721cellsapoptosision and cells division arrestion.