Effect of microRNA-21 inhibitor combined with graphene quantum dots loaded cisplatin nano-drug system on the apoptosis,migration and invasion of melanoma cells
Objective To investigate the effect of microRNA-21(miRNA-21)inhibitor combined with graphene quantum dots loaded cisplatin nano-drug system(GPt)on the apoptosis,migration and invasion of melanoma cells and its mechanism.Method B16F10 melanoma cells were cultured and divided into control group,nano-drug 24 h group,nano-drug 48 h group and nano-drug 72 h group.Methyl thiazolyl tetrazolium(MTT)assay was used to screen the appropriate dose and administration time of nano-drug;then they were divided into miRNA-21 inhibitor group,nano-drug group,miRNA-21 inhibitor+nano-drug group.Transwell assay was used to detect cell invasion ability,scratch test was used to detect cell migration ability,flow cytometry was used to detect cell cycle and apoptosis.Polymerase chain reaction(PCR)and Western blot were used to detect the expression of miRNA-21,B cell lymphoma/leukemia-2(Bcl-2),p53 and Bcl-2 associated X protein(BAX).And the B16F10 cells were subcutaneously injected into nude mice to establish a melanoma model,and the tumor weight of each group were detected.Result The screened time and dose of nano-drug were 72 h and 5.0 μl,respectively,miRNA-21 inhibitor had obvious interference effect,which could be used for later experiments.Compared with the control group,the invasion and migration ability of the cells in the nano-drug group and the miRNA-21 inhibitor group were decreased,the proportion of cells in G1 phase was increased,the proportion of cells in S phase was decreased,and the proportion of apoptosis cells was increased.The effect of the combination of the two groups was more obvious.The relative expression of Bcl-2 protein and mRNA in the miRNA-21 inhibitor group and the nano-drug group decreased,and the decrease was more obvious after the combination of the two groups,the expression of p53 and BAX protein and mRNA increased,and the increase was more obvious after the combination of the two groups.Compared with the control group,the tumor weight of the other groups decreased,and the miRNA-21 inhibitor+nano-drug group de-creased most significantly.In the control group,the tumor tissue was rich in blood vessels,the tumor cells were closely ar-ranged,and grew actively and were relatively uniform in size.In the other groups,the tumor cells were loosely arranged and the cell growth was not active.Conclusion GPt and miRNA-21 inhibitor can induce apoptosis and inhibit the migra-tion and invasion of melanoma cells.
melanomamicroRNA-21graphene quantum dots loaded with cisplatin nano-drug systemmechanism
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