Preliminary Study On The Inhibitory Effect Of Methyl Jasmonate Combined With Cisplatin On The Proliferation Of Human Tongue Cancer Cell CAL-27

Background:Tongue cancer is the most common oral squamous cell carcinoma,which is often treated by surgery combined with drugs.However,at present,traditional chemotherapy drugs,such as cisplatin(CDDP),mostly have serious adverse reactions,which aggravate patients’ pain.It is of great significance to explore new natural bioactive drugs that can inhibit tumor proliferation from the perspective of improving the therapeutic effect of anti-tumor and reducing the side effects.Methyl jasmonate(MEJA),as a natural plant hormone derivative,has been proved to be able to inhibit the proliferation of a variety of tumor cells by a number of domestic and foreign studies,with small side effects,and has a broad application prospect in cancer prevention and treatment.In this study,MEJA combined with CDDP was used to investigate the inhibitory effect of MEJA on the proliferation of human tongue cancer cell line CAL-27.By confirming the anti-tumor effect of MEJA,we can provide a new idea for MEJA in the treatment of tongue cancer.Objective:1.To study the inhibitory effect of MEJA combined with CDDP on the growth of tongue cancer cell line CAL-27.2.To investigate the mechanism of MEJA combined with CDDP inducing apoptosis of tongue cancer cell line CAL-27.Methods:1.According to the design,CAL-27 cells were divided into four groups:blank control group,MEJA group,CDDP group,MEJA+CDDP combination group;MEJA and CDDP of different concentrations were used to act on CAL-27 cells in different time,alone and in combination;cell morphology was observed by inverted microscope;2.CCK-8 method was used to detect the proliferation inhibition rate of CAL-27 cells in each group,and the appropriate time and concentration of drug action were selected;3.Annexin V FITC/PI double staining was used to detect the level of apoptosis in each group;4.The mRNA expression of Bcl-2,Caspase-3 and other tumor related genes in CAL-27 cells was detected by PCR.Results:1.Under the inverted microscope,it s observed that the cells in the blank control group are in good condition,while the CAL-27 cells in the MEJA group and CDDP group is exfoliated and disintegrated,the adherent cells is less,the suspension cells is smaller and rounder,the cell spacing is increased,and the intracellular particles is increased.In the combination group,the above morphological changes of the cells are more significant;2.CCK-8 test result shows that:compared with the blank control group,the proliferation inhibition rate of CAL-27 cells in MEJA group and CDDP group are increased;with the increase of concentration gradient,the proliferation inhibition rate of MEJA group and CDDP group raise approximately the same;while the proliferation inhibition rate of combination group is higher than that of single drug group;there are corresponding dose effect and time effect in each group;3.The results of flow cytometry shows that the apoptosis rates of CAL-27 cells in MEJA group and CDDP group are higher than that in the control group,while the apoptosis rate of CAL-27 cells in combination group is significantly higher than other groups;4.PCR results shows that compared with the control group,the expression levels of Bcl-2 decreased and caspase-3 are increased in MEJA group and CDDP group;this is more obvious in the combination group.Conclusion:MEJA combined with CDDP can inhibit the proliferation of CAL-27 cells and induce its apoptosis.The mechanism is that MEJA can inhibit the proliferation and apoptosis of CAL-27 cells by inducing the down-regulation of Bcl-2 expression level and the up-regulation of Caspase-3 expression level.