Research On Gastric Cancer Anticancer Activity Of Cantharidin Acid Magnesium In Vitro And Its Mechanism

Objective: To compare the anticancer activity of Cantharidin acid magnesium which is self-developed with cantharidin and Cantharidin acid sodium in vitro,screening out the best anti-cancer drug,and to explore its underlying mechanisms.Methods: 1.The proliferation inhibition of human gastric cancer MKN-28,SGC-7901,BGC-823 cells which cultured in different concentrations of Cantharidin acid magnesium,cantharidin,Cantharidin acid sodium method in vitro,was detected by CCK-8(Cell Counting Kit-8);2.The cell morphological changes of the three gastric cancer cells cultured in different concentrations of Cantharidin acid magnesium,cantharidin,Cantharidin acid sodium was observed by inverted phase contrast microscope;3.The human normal gastric mucosal GES-1 cells was cultured as the control group to evaluate the cytotoxicity of Cantharidin acid magnesium.4.Transmission electron microscope(TEM)was used to observed the ultrastructural changes of Cantharidin acid magnesium on(on)human gastric cancer BGC-823 cells.5.The cell cycles and apoptosis of Cantharidin acid magnesium on human gastric cancer BGC-823 cells was tested by flow cytometry.6.The expression of CDK1,cyclin B1 protein was analyzed by western blot.Results: 1.Cantharidin acid magnesium,cantharidin,Cantharidin acid sodium have inhibitory effect on three gastric cancer cells of MKN-28,SGC-7901,BGC-823,and showed a dose-response relationship(P<0.05),and the inhibitory effect of magnesium cantharidin is the best;2.inverted phase contrast microscope : After the cantharidin acid magnesium,cantharidin,Cantharidin acid sodium acting on the three gastric cancer cells 24 h,the number of cells decreased gradually,and the phenomenon of cell wrinkle and shrink,becoming smaller and round,some cells disruption were induced;3.The gastric cancer BGC-823 cells had shown Obvious inhibition under the action of the Cantharidin acid magnesium in 0.52μmol / L,While the inhibition rate of normal gastric mucosa GES-1 cells was still low at 1.04μmol / L.The inhibitory effect of Cantharidin acid magnesium on gastric cancer cell line BGC-823 was stronger than that of human normal gastric mucosa GES-1 under the same drug concentration.Their half inhibitory concentration(IC50)was 4.868μmol / L and9.645μmol / L respectively.4.TEM observation: after using half maximal inhibitory concentration of Cantharidin acid magnesium for twenty-four hours,BGC-823 cells’ nuclear represented abnormity,cleavage and fragmentation,endoplasmic reticulum and mitochondria swelling;5.flow cytometry results showed that: Afer the human gastric cancer BGC-823 cells was in the role(under the action)of Cantharidin acid magnesium,the cell proportion was decreased in G0/G1 phase,increased in G2/M phase,and unchanged basically in S phase,indicated that the cell appears G2/M phase arrest(P<0.05);Annexin V-FITC/PI double staining showed that the cell apoptosis rate was increased along with the magnesium cantharidate concentration increased gradually(P<0.05)6.Western-Blot results showed that: after using Cantharidin acid magnesium for Twenty-four(24)hours,The expression of CDK1 and cyclin B1 decreased significantly on gastric cancer BGC-823 cells compared with the blank group(**P<0.05,##P<0.05).Conclusion: 1.The inhibitory effect of Cantharidin acid magnesium on gastric cancer cells line is better than cantharidin and Cantharidin acid sodium;2.Cantharidin acid magnesium can inhibit the proliferation of human gastric carcinoma BGC-823 cells,and significantly block the BGC-823 cells in the G2/M phase,and induce BGC-823 cells apoptosision(apoptosis).It may be related to down-regulation of the expression of CDK1 and Cyclin B1 protein.3.The toxicity of Cantharidin acid magnesium on anti-gastric cancer cell activity is cell selective and sensitive in a certain degree.