Effect and Mechanism Study of Rose roxburghii Tratt.Fruits Polysaccharide on Proliferation and Apoptosis of Human Lung Cancer A549 Cells
Objective To investigate the effect and mechanism of Rose roxburghii Tratt.fruits polysaccharide(RTFP)on proliferation and apoptosis of lung adenocarcinoma A549 cells.Methods After A549 cells were treated with 0,2,4,6,8,10 g·L-1 RTFP for 24,48,72 hours,the proliferation of A549 cells was detected by CCK-8 method.After A549 cells were treated with 0,2,6 and 10 g·L-1 RTFP for 24 and 48 hours,the migration of A549 cells was determined by cell scratch test.Transwell assay was used to determine the invasion of A549 cells.The cell cycle of A549 cells was determined by flow cytometry(PI single staining method)after A549 cells were treated with 0,2,6 and 10 g·L-1 RTFP for 48 hours.The apoptosis of A549 cells was detected by flow cytometry(Annexin V-FITC/PI double staining).The expression levels of cell cycle and apoptosis-related genes in A549 cells were determined by Real-Time PCR.The expression levels of cell cycle and apoptosis-related proteins in A549 cells were determined by Western Blot.Results(1)After 24,48 and 72 hours of intervention,compared with the control group(0 g·L-1),the growth of A549 cells in the RTFP 2,4,6,8 and 10 g·L-1 groups was significantly inhibited,and the survival rate of cells was significantly decreased(P＜0.05,P＜0.01).The IC50 value of RTFP intervention for 48 hours was 6.109 g·L-1,and the concentration of 2,6 and 10 g·L-1 was used as the low-,medium-and high-dose groups of RTFP in subsequent experiments.(2)After 48 hours of intervention,compared with the control group,the migration distance of A549 cells in the RTFP 2,6 and 10 g·L-1 groups was significantly increased(P＜0.05,P＜0.01),and the number of cell invasion was significantly decreased(P＜0.05,P＜0.01).The proportion of cells in G0/G1 phase and G2/M phase was significantly increased(P＜0.05,P＜0.01),and the proportion of cells in S phase was significantly decreased(P＜0.01)in the 6 and 10 g·L-1 groups.The mRNA and protein expressions of G0/G1 and G2/M phase-related regulatory factors CDK-4,CDK-6,CyclinD1,CDK-1 and CyclinB1 of A549 cells in the 2,6 and 10 g·L-1 groups of RTFP were significantly increased(P＜0.05,P＜0.01).The apoptosis rate of A549 cells was significantly increased(P＜0.01).The mRNA expressions of apoptosis-related genes Caspase-3,Caspase-8 and Caspase-9 were significantly up-regulated(P＜0.05,P＜0.01).The protein expressions of pro-apoptotic Bax,Caspase-3,Caspase-8 and Caspase-9 were significantly up-regulated(P＜0.05,P＜0.01),and the protein expressions of anti-apoptotic Bcl-2 was significantly down-regulated(P＜0.05,P＜0.01).Conclusion RTFP could inhibit the proliferation,migration and invasion of A549 cells,block the cell cycle in G0/G1 and G2/M phases,and induce apoptosis of A549 cells through mitochondrial pathway and death receptor pathway.
Rose roxburghii Tratt.fruits polysaccharide(RTFP)lung cancer A549 cellscell proliferationmigrationinvasioncell cycleapoptosis
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