The Study Of Poly(I:C) On Non-small Cell Lung Cancer LL/2 Cells Inhibition Of Proliferation?Invasion And Migration

Objective:To investigate the effects of poly(I:C)complex on proliferation,invasion and migration of LL/2 cell line in vitro.Methods:The cells lines were divided into blank control group(NT group)and poly(I:C)complex group.The effects of poly(I:C)complex at different concentration and time on the viability of LL/2 cells were detected by cell proliferation-toxicity assay.The effects of NT group and poly(I:C)complex group on the doubling time of LL/2 cells were detected by single layer cell proliferation assay.Annexin V-FITC/PI double staining was used to detect the apoptosis induction of LL/2 cells in NT group and poly(I:C)complex group.Flow cytometry was used to detect the effect of NT group and poly(I:C)complex group on LL/2 cell cycle arrest.Real-time PCR indicated that the mRNA expression of TLR3 was significantly up-regulated in poly(I:C)group compared with NT group,which was statistically significant(P<0.001).Transwell migration assay was used to detect the effect of NT group and poly(I:C)complex group on the longitudinal movement and invasion ability of LL/2 cells.The effect of NT group and poly(I:C)complex group on the lateral motility of LL/2 cells was detected by scratch assay.Results:1.Cell proliferation-toxicity experiment results:LL/2 cells were significantly inhibited by poly(I:C)complexes at different concentrations and time points at 10 ug/ml,50 ug/ml,100 ug/ml,and 200 ug/ml,showing a dose-time dependence.2.Monolay cell proliferation experiment results:The total number of LL/2 cells in the Poly(I:C)complex group was significantly lower than that in the NT group at 24h and 48h after the intervention,with statistical significance(24h P<0.01,48h P<0.001).3.Annexin v-fitc/PI double staining results:Compared with the NT group,the apoptosis rate of LL/2 cells in the poly(I:C)complex group decreased significantly after 48h,suggesting that the late apoptosis could be significantly induced(P<0.001).4.Flow cytometry results:The proportion of LL/2 cells in the poly(I:C)complex group was significantly higher than that in the NT group after 48h(P<0.001),and the proportion of the corresponding S and G2 cells was significantly lower(P<0.005 in the S phase and P<0.001 in the G2 phase).5.Real-time PCR results:The mRNA expression of TLR3 was significantly up-regulated in poly(I:C)group compared with NT group,which was statistically significant(P<0.001).6.Transwell results:The Poly(I:C)complex group showed a statistically significant decrease in both longitudinal motion and vertical invasion compared with the NT group(P<0.01).7.Scratch test results:The surface area of the Poly(I:C)complex group was wider at 24h and 48h than that of the NT group,suggesting a shorter migration distance(24h P<0.05,48h P<0.01).Conclusions:1.In vitro experiments,Poly(I:C)complex promoted apoptosis and inhibited proliferation of LL/2 cells,suggesting that Poly(I:C)complex induced apoptosis of lung cancer cells and played an anti-tumor growth role.2.Poly(I:C)complex inhibited LL/2 cell migration,invasion and parallel movement in vitro.