The Effect Of Polysaccharides From Metaplexis Japonica Makino Nutshell On The Apoptosis And Migration Of Human Lung Cancer A549 Cells

ObjectiveMetaplexis japonica is a perennial herb whose husk has multiple biological functions.At present,the research on the anti-tumor activity of polysaccharides from Metaplexis japonica Makino nutshell(MJP-1’)is still in its initial phase.In this experiment,the effect of MJP-1’ on the apoptosis and migration of human lung cancer A549 cells were used to explore its anti-tumor activity.MethodsThe effect of MJP-1’ on the viability of A549 cells was detected using CCK8 assay.The release of LDH in A549 cells was detected using colorimetry.The effect of MJP-1’ on the apoptosis of A549 cells was explored by TUNEL assay,while those on the mitochondrial membrane potential and ROS of A549 cells were investigated using JC-1 and DCFH-DA fluorescent probes.Moreover,the expressions of apoptosis-related proteins in A549 cells were measured using Western blot.The effect of MJP-1’ on the migration of A549 cells was preliminarily determined using wound healing assay,Quantitative Real-time PCR,Western blot and immunofluorescence assay.ResultsThe CCK8 assay showed that MJP-1’ inhibited the proliferation of A549 cells in a dose-dependent manner.Colorimetry presented an increase of LDH release in A549 cells.TUNEL assay revealed that MJP-1’ could promote the apoptosis of A549 cells.The JC-1 probe experiment demonstrated that MJP-1’induced the reduction of mitochondrial membrane potential in A549 cells.The DCFH-DA probe experiment showed that as the concentration of MJP-1’increases,the level of ROS gradually increased.The Western Blot results exhibited that MJP-1’ could down-regulate the expression of Bcl-2 protein and increase the expressions of Bax and cleaved-caspase-3 proteins.According to wound healing assay,MJP-1’ could inhibit the migration of A549 cells in a dosedependent manner.QPCR results showed that MJP-1’ could reduce the RNA expressions of HMGB1,OPN and Rho A,and Western blot results presented that MJP-1’ could decrease the protein expressions of HMGB1,OPN and Rho A.Based on immunofluorescence assay results,it was found that Rho A protein was localized in the nucleus and decreased with the increase of drug concentration.ConclusionsThis study reveals that MJP-1’ can induce the apoptosis of human lung cancer cells A549,and inhibit the migration of A549 cells by reducing the expressions of HMGB1,OPN and Rho A.The experiment provides a theoretical basis for the use of natural polysaccharides as a candidate for new anti-tumor therapies.