Effect of Shenmai injection on chemotherapy resistance of gastric cancer cells and underlying mech-anism
Objective To explore the underlying mechanism of how shenmai injection regulate the multidrug resistance of gastric cancer cells by regulating microRNA-140-3P(miR-140-3P).Methods The human gastric cancer multidrug-resistant cell line HGC-27/DCF(5-Fu,cisplatin and docetaxel)cells(purchased from the Cell Culture Collection,USA)were established and cultured.These cells were divid-ed into following groups:control group(DCF+saline),negative control group(DCF+miR-140-3p inhib-itor NC+saline),shenmai injection group(DCF+miR-140-3p inhibitor NC+shenmai injection injec-tion),miR-140-3p inhibitor group(DCF+miR-140-3p inhibitor+saline),and shenmai injection+miR-140-3p inhibitor group(DCF+miR-140-3p inhibitor+shenmai injection injection).The levels of miR-140-3p,PD-1,PD-L1 were determined by RT-qPCR and western blot assays.The proliferation levels of HGC-27/DCF cells were determined by cell proliferation and toxicity assay kit(CCK-8).The apoptosis rate of HGC-27/DCF cells were determined by annexin V and propidium iodide(PI).The cell invasion as-say(Transwell)assay was adopted to determine the cell invasion ability.One-way analysis of variance(ANOVA)was applied for comparison between groups.Results The expression levels of miR-140-3p in GES-1,HGC-27,and HGC-27/DCF cells were 3.02±0.67,1.04±0.17,and 0.26±0.08,respectively(F=33.800,P＜0.05),the expression levels of PD-1 were 0.21±0.04,0.30±0.05,0.59±0.08,re-spectively(F=33.800,P＜0.05),and expression levels of PD-L1 were 0.09±0.02,0.16±0.03,0.39±0.05(F=58.355,P＜0.05),respectively;post hoc two-by-two comparisons showed that miR-140-3p expression levels were higher than those in HGC-27/DCF cells than in GES-1 and DCF cells,PD-1 and PD-L1 were expressed at lower levels in HGC-27/DCF cells than those in GES-1 and HGC-27 cells,and the differences between the groups were statistically different(P＜0.05).The cell proliferation viability in the control group,negative control group,miR-140-3p inhibitor group,shenmai injection group,and shenmai injection+miR-140-3p inhibitor group were 0.47±0.03,0.48±0.02,0.30±0.04,0.68±0.11,and 0.45±0.04,respectively(F=16.565,P＜0.05),and the apoptosis rate were 8.52± 0.75,9.02±0.66,21.52±4.25,3.88±0.55,and 15.63±3.02(F=25.014,P＜0.05),respective-ly;post hoc two-by-two comparisons showed that the cell proliferation viability of the control group was higher than that of the miR-140-3p inhibitor group but lower than that of the shenmai injection group,and the cell proliferation viability of the shenmai injection+miR-140-3p inhibitor group was higher than that in miR-140-3p inhibitor group but lower than shenmai injection group,and the difference between the groups was statistically significant(P＜0.05);the apoptosis rate of the control group was lower than that of miR-140-3p inhibitor group but higher than that of shenmai injection group,and the apoptosis rate of shen-mai injection+miR-140-3p inhibitor group was lower than that of miR-140-3p inhibitor group but higher than that of shenmai injection group,and the difference between the groups was statistically different(P＜0.05).The numbers of cell invasion in the control group,negative control group,miR-140-3p inhibitor group,shenmai injection group,and shenmai injection+miR-140-3p inhibitor group were 162.50± 25.20,155.80±18.30,208.80±36.50,123.50±25.60,and 166.40±18.50,respectively(F=4.228 P＜0.05);post hoc two-by-two comparisons showed that the number of cell invasion in the control group was lower than that of the miR-140-3p inhibitor group but higher than that of the shenmai injection group,and the number of cell invasion in the shenmai injection+miR-140-3p inhibitor group was lower than that of the miR-140-3p inhibitor group but higher than that of the shenmai injection group,with a sta-tistically significant difference between the groups(P＜0.05).Conclusion The effect of shenmai injec-tion could be antagonized by miR-140-3p inhibitor,and shenmai injection could mitigate the DCF multi-re-sistance of gastric cancer cells by regulating the miR-140-3p-mediated PD-1/PD-L1 signaling pathway.
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