Study of effects and mechanism of cinobufotalin on non-small cell lung cancer cells
Objective To explore the mechanism of cinobufotalin regulating phosphatase and tensin homolog(PTEN)and member of wingless MMTV integration site family(Wnt)signaling pathway through microRNA-21(miR-21)on non-small cell lung cancer(NSCLC)cells.Methods HCC827 cells were divided into control group(normal culture),inhibitor-NC group(transfected with miR-21 inhibitor negative control),miR-21 inhibitor group(transfected with miR-21 inhibitor),cinobufacini(Cin)group(0.15 µg·mL-1 cinobufacini),miR-21 mimics group(0.15 µg·mL-1 cinobufacini+miR-21 mimics),OE-PTEN group(0.15 μg·mL-1 cinobufacini+miR-21 mimics+overexpression PTEN plasmid).The mRNA expression levels programmed death ligand-1(PD-L1)were detected by quantitative real-time polymerase chain reaction(qRT-PCR);the number of cell invasion was detected by Transwell chamber method;the expression levels of Wnt/β-catenin pathway-related proteins were detected by Western blot.Results The number of cell invasion in the control group,Cin group,miR-21 mimics group and OE-PTEN group were 121.16±16.43,68.56±11.21,149.30±21.26 and 96.13±13.31,respectively;the protein expression levels of Wnt4 were 1.00±0.13,0.45±0.06,0.93±0.12 and 0.55±0.08,respectively;the protein expression levels ofβ-catenin were 1.00±0.14,0.26±0.04,0.82±0.10 and 0.15±0.03;the mRNA expression levels of PD-L1 were 1.00±0.08,0.71±0.09,1.21±0.11 and 0.70±0.13,respectively.The above indicators of the control group were compared with the Cin group,the above indicators of the Cin group were compared with the miR-21 mimics group,and the above indicators of the miR-21 mimics group were compared with the OE-PTEN group,the differences were statistically significant(all P＜0.05).Conclusion Cinobufotalin may improve the immune function of NSCLC cells,inhibit their invasion and proliferation by down-regulating miR-21 expression to regulate PTEN,Wnt/β-catenin pathway.
cinobufotalinnon-small cell lung cancer cellsphosphatase and tensin homologsβ-cateninproliferationinvasion
万方数据
维普
