| Background:Osteosarcoma is one of the most common malignant tumors in modern medicine and clinical practice in the world.At present,its treatment method is mainly surgery combined with drug chemotherapy.However,traditional chemotherapeutic drugs have many side effects and the multidrug resistance of osteosarcoma to chemotherapeutic drugs also limits the clinical application.Therefore,it is of great significance to find new low-toxicity natural anti-tumor drugs.Although cisplatin is a commonly used chemotherapeutic drug in the clinical treatment of osteosarcoma and has a good effect,the side effects of CDDP are relatively large,the most important of which is nephrotoxicity.Tea culture has thousands of years of cultural history in my country.The main active ingredient in tea is epigallocatechin gallate,which is one of the main active substances in catechins.It is a kind of amphiphilic molecule that is both hydrophilic and lipophilic.It contains a variety of active genes,includes anti-tumor,anti-oxidation and anti-inflammatory effects.It can interact with transcription growth factors,protein kinases,cytokines and enzymes and other molecular targets.Combining and interacting,leading to apoptosis of cancer cells,EGCG is an ideal class of potential anticancer drugs.Studies have reported that EGCG can reduce the damage of CDDP to cells by improving the antioxidant capacity of cells.Thus,this experiment intends to use ECGC combined with CDDP to act on human osteosarcoma MG-63 cells to preliminarily explore its inhibitory effect on the proliferation of MG-63 cells and its mechanism.Methods:(1)Different concentration gradients of EGCG ester and CDDP were used separately or in combination for MG-63 cells.The experiment is divided into 4groups: blank control group,EGCG group,CDDP group and EGCG combined with CDDP group.(2)inverted phase contrast microscope Observe cell morphological changes under the following conditions.(3)Using CCK-8 method to detect cell growth inhibition rate.(4)Using Transwell chamber method to detect cell invasion number and wound healing test is used to detect cell migration ability.(5)Western blot detection of related proteins Bax,Bcl-2,Bcl-x L,MMP-2,MMP-9,Caspase-3,cleaved Caspase-3 expression.Results:(1)We can see that the blank control group MG-63 cells grow well.The cells in the other experimental groups at different concentrations have different cell morphologies and reduced numbers.Different numbers of cells can be seen falling off,the density of adherent cells is reduced,and the cell morphology becomes round and smaller.The number of necrotic cells in the CDDP combination group was greater than that in the single-drug group,and these were of time-dependent and drug concentration-dependent.(2)CCK-8 method results show that different concentrations of EGCG and CDDP can inhibit the proliferation of osteosarcoma MG-63 cells to varying degrees,resulting in a decrease in cell viability,and the inhibitory effect on the cells becomes more significant with the increasing of drug concentration.The combined drug group had stronger inhibitory effect on the growth of MG-63 cells(P<0.05);(3)Transwell chamber method showed that the number of cells passing through the compartment membrane decreased in both the EGCG group and the CDDP group,and the invasion ability of MG-63 cells was weakened;The number of cells passing through the compartment membrane in the EGCG+CDDP group decreased more significantly,indicating that the combined medication had the strongest inhibitory effect on the invasion ability of MG-63 cells;The wound healing test showed that the migration ability of MG-63 cells in the EGCG group and the CDDP group were both decreased,the cell migration ability of the combined drug group weakened more significantly,indicating that EGCG combined with CDDP can effectively inhibit the migration and invasion of osteosarcoma MG-63 cells(P<0.05);(4)Western blot results showed that the expression levels of Bcl-2,Bcl-x L,MMP-2 and MMP-9 were significantly down-regulated,and the levels of Bax,Caspase-3,and cleaved Caspase-3 were significantly up-regulated.Compared with single-drug application,the combined drug group MG-63 cells have the highest inhibitory ratio,and their related protein expression levels are the most up-regulated/down-regulated Conclusion:EGCG can inhibit the growth of osteosarcoma MG-63 cells,combined with CDDP can effectively inhibit its proliferation,invasion and migration ability,and induce cell apoptosis,possibly by down-regulating Bcl-2,Bcl-x L,MMP-2,MMP-9and up-regulate the expression of Bax,Caspase-3,and cleaved Caspase-3 to inhibit the growth of MG-63 cells.The results of this study provide a new theoretical basis for the clinical treatment of osteosarcoma. |
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