| Objective:To investigate the effect of methylselenic acid(MSA)and cisplatin(DDP)on the chemosensitivity of oral cancer cells in order to provide theoretical basis for enhancing the chemosensitivity of oral cancer.Methods:1 Oral cancer Tca8113 and KB cell lines were selected and divided into control group and MSA group.The following experiment was performed.(1)Cell proliferation was detected by MTS assay and cell cloning assay;(2)Cell migration ability was detected by cell scratch assay;(3)Apoptosis rate and cell cycle were detected by flow cytometry.2.Oral cancer Tca8113 cell line was selected.Divided into control group,MSA group,DDP group,combined group,the following experiment.(1)The cell proliferation were detected by MTS method;the morphological changes were observed by inverted microscope;(2)Apoptosis rate and cell cycle were detected by flow cytometry;(3)Detection of apoptosis by DAPI staining;(4)The level of reactive oxygen species in each group was detected by fluorescent probe DCFH-DA;(5)The Caspase-3 enzyme in each group was detected by fluorescence microplate reader.Results:1.Study on the effect of MSA on oral cancer cells(1)The half-inhibitory concentration(IC50)of MSA against Tca8113 and KB cells was 0.2μM.MTS results showed that MSA significantly inhibited the proliferation of oral cancer cells,and the inhibition was concentration-dependent;cell cloning experiments showed that MSA inhibited the clonality of oral cancer cells(P<0.001);0.1 μM MSA has little effect on the proliferation of oral cancer cells,so we chose 0.1 μM MSA as the second part of the experimental dose combined with cisplatin;(2)Cell scratch test results showed that MSA inhibits oral cancer cells Migration(P<0.001);(3)Annexin V-FITC/PI double staining showed that MSA had obvious pro-apoptotic effect;(4)PI single-stain flow method showed that MSA had S phase cell arrest.2.Study on the effect of DDP combined with MSA on oral cancer(1)MTS results showed that both MAS and DDP could inhibit the growth of oral cancer cells,and MSA could promote the growth inhibition of DDP on oral cancer cells and increase the sensitivity of DDP chemotherapy.(2)Inverted fluorescence microscopy showed that compared with the control group,the number of cells in the experimental group decreased and the morphology was irregular,especially in the combined group;(3)Annexin V-FITC/PI double staining flow results showed that compared with the control group,MSA group,DDP group,combined group increased the apoptotic rate(P<0.001)Among them,the increase of apoptosis was the most significant in the combination group;(4)DAPI staining showed that compared with the control group,apoptotic cells were observed in MSA group,DDP group and combination group,which showed dense staining,nuclear fragmentation and nuclear pyknosis,and the combined group was the most obvious;(5)The indirect fluorescent labeling method showed that compared with the control group,the ROS levels in the MSA group,the DDP group and the combined group were increased(P<0.05),and the combined group was the most obvious;(6)The fluorescence microplate reader showed that Compared with the control group,the activities of Caspase-3 in the MSA group,the DDP group and the combined group were increased(P<0.001),and the combined group was the most significant.Conclusion:1.MSA inhibits the growth,proliferation and migration of oral cancer cells,induces cycle arrest and promotes apoptosis..2.0.1 μM MSA can enhance the sensitivity of oral cancer cells to DDP chemotherapy.3.The mechanism may be as follows:(1)inhibiting the antioxidant system,increasing ROS accumulation in cells,promoting DNA damage;(2)inducing cell cycle arrest,preventing the repair of damaged DNA,inhibiting cell proliferation;these aspects interact with oral cancer cells and ultimately promote cell apoptosis. |
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