Molecular mechanism of circular RNA C190 promoting the progression of non-small cell lung cancer
Objective To investigate the effect of circular RNA(circRNA)C190 on the progres-sion of non-small cell lung cancer and its molecular mechanism.Methods Total RNA was extracted from human normal lung epithelial cells(BEAS-2B)and non-small cell lung cancer NCI-H358,NCI-H1299,A549 and NCI-H1568 cells,and the expression level of circRNA C190 in cells was analyzed by fluores-cence quantitative polymerase chain reaction(PCR).NCI-H1299 cells with the most significant difference in the expression were selected as the research objects,and RNA interference technology was used to estab-lish control,circRNA C190 knockdown and circRNA C190 overexpression cell lines,which were control group,circRNA C190 KD group and circRNA C190 OE group,respectively.Cell viability of the three groups was analyzed by cell counting experiment.Cell cycle and apoptosis levels of the three groups were analyzed by flow cytometry.Subcutaneous tumor formation ability of the three groups was analyzed by in vitro xenograft experiment.Target genes of circRNA C190 were analyzed by bioinformatics and dual lucifer-ase reporter gene.Comparison of quantitative data between groups was performed by t test.Results The expression level of circRNA C190 in human normal lung epithelial cells(BEAS-2B)(0.99±0.10)was significantly lower than that in non-small cell lung cancer NCI-H358,NCI-H1299,A549 and NCI-H1568 cells(1.43±0.14,1.90±0.17,1.48±0.10,1.40±0.10,t=6.349,11.440,8.779,7.070,P＜0.05).The absorbance value of the control group(1.45±0.14)was significantly higher than that of the circRNA C190 KD group(1.04±0.13,t=4.834,P＜0.05).The absorbance value of control group(1.45±0.14)was significantly lower than that of circRNA C190 OE group(1.84±0.09,t=5.257,P＜0.05).The tumor volume and tumor weight of control group[(502.83±63.84)mm3,(4.43±0.49)g]were significantly greater than those of circRNA C190 KD group[(352.33±44.72)mm3,(1.70± 0.34)g,t=4.729,11.300,P＜0.05].The tumor volume and tumor weight of control group[(502.83±63.84)mm3,(4.43±0.49)g]was significantly lower than those of circRNA C190 OE group[(671.50±52.29)mm3,(7.61±0.80)g,t=5.006,8.326,P＜0.05].The proportion of Go/G1 phase of control group cells[(44.09±2.79)％]was significantly lower than that of circRNA C190 KD group cells[(54.44±2.54)％,t=6.711,P＜0.05].The proportion of G0/G,phase of control group cells[(44.09±2.79)％]was significantly higher than that of circRNA C190 OE group cells[(33.32± 2.05)％,t=7.611,P＜0.05].The apoptosis rate of control group cells[(4.77±0.78)％]was signifi-cantly lower than that of circRNA C190 KD group cells[(14.36±2.40)％,t=9.333,P＜0.05].The absorbance value of control group cells[(4.77±0.78)％]was significantly lower than that of circRNA C190 OE group cells[(1.59±0.93)％,t=6.405,P＜0.05].CircRNA C190 is the sponge of miR-200c-3p.Conclusion The expression level of circRNA C190 is increased in lung cancer cells.Knockdown of circRNA C190 significantly inhibits cell proliferation and migration,while overexpression of circRNA C190 promotes cell proliferation and migration,mainly through binding miR-200c-3p to exert bio-logical effects.
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