The Inhibition Effect Of Paeony Total Glucosides On Non-small Cell Lung Cancer Through Influencing The Expression Of MMP-9 And HIF-1α

The paeonol total glucosides(TPG)have anti-tumor effection,it is still unknown whether the effect is related to matrix metalloproteinase-9(MMP-9)and hypoxia-inducible factor-la(HIF-1a).In the present study,effects of total glucosides of paeony on the expression of MMP-9 and HIF-1α in the H1299 cellsand tumor model mouse were studied,and the related molecular mechanisms were discussed.Inhibitory effect of TPG on the growth of H1299 cells was analyzed by MTT colorimetric assay,while the expression veldsof MMP-9 and HIF-1α mRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR).The expression levels of MMP-9 and HIF-1α protein in H1299 cells were studied by VWestern blotting.In vitro study showed that the growth of lung cancer cells was inhibited by TPG in a dose-dependent manner at 24 hours,48 hours and 72 hours(P<0.05).The mRNA level and protein level of MMP-9 and HIF-1α in lung cancer cell H1299 was decreased in a dose-dependent manner afte co-culture with TPG for 24 hours.C57BL/6 lung cancer model mice were divided randomly into five groups,as model control group,cyclophosphamide group(0.06 g/kg),TPG small dose group(0.06 g/kg),TPG medium dose group(0.24 g/kg)and TPG high dose group(0.96 g/kg).Levels of MMP-9 and HIF-1α in serum were measured by enzyme-linked immunosorbent assay(ELISA)after administration with the corresponding drugs for 4 weeks,RT-PCR was used to analyze the MMP-9 and HIF-1α mRNA expression.Protein levels of MMP-9 and HIF-1α in tumor tissue were detected by Western blotting.Results of in vivo test showed that tumor tissues volume in model control group C57BL/6 mouse kept growing after model establishment.The tumor volumes of C57BL/6 mice in medium dose group and in high dose group were significantly different from those in model control group.At the 3rd day,1st week,2nd week,3rd week and 4th week,the inhibitory rates of medium dose TPG on the tumor volume were 15.20%,26.05%,24.10%,21.59%and 24.71%,those of high dose TPG were 32.80%,31.51%,55.00%,37.28%and 39.64%,respectively.At the 4th week after model establishment,the tumor weight of C57B/L mice in TPG medium dose group and in TPG high dose group were significantly decreased compared with those in model control group(P<0.05);the tumor weight of C57B/L mice in TPG high dose group were significantly decreased compared with those in TPG small dose group and medium dose group(P<0.05);the inhibitory rates of medium dose TPG and high dose TPG were respectively 31.43%and 43.47%.The expression levels of MMP-9 and HIF-1α in blood and in tumor tissues,both in TPG medium dose group and in TPG high dose group,were decreased(P<0.05).There was no significant difference between TPG small dose group and model control group.In conclusion,TPG could inhibit the growth of H1299 lung cancer cell in vitro and in vivo.The anti-tumor effects could be explained by down-regulating the expression of the two key genes MMP-9 and HIF-1α.