| Purpose:To observe the MMP-9 of Human Lung Adenocarcinoma Cells A549 expression changes and to explore the effect of Serum Containing Buzhong Yiqi Decoction in Invasion and Metastasis of lung Adenocarcinoma Cells A549 by using Buzhong Yiqi Decoction intervent the Human Lung Adenocarcinoma Cells A549 mediated by TGF-β1. We can provide objective experimental evidence for the Peitu Shengjin Chinese medicine of Buzhong Yiqi Decoction in the therapy of lung cancer and reversion multidrug resistance of lung cancer chemotherapy.Material and method:The serum containing Buzhong Yiqi decoction was prepared by adopting the method of serum pharmacology. 40 rats were divided into the four groups:high(11.34 g·kg-1), middle(5.67 g·kg-1), low(2.84 g·kg-1) dose groups of Buzhong Yiqi Decoction and normal saline(10 ml·kg-1). At the end of the 3thday serum was obtained. The groups were: 10% blank serum group, TGF-β1 mediated group, and 10% high, middle and low dose groups of serum Buzhong Yiqi Decoction+TGF-β1 group.The test of morpgology changes Cells A549 mediated by TGF-β1 was by cytomorphology. Inhibition rate of Buzhong Yiqi Decoction on MMP-9 and Integrinβ1 of Human Lung Adenocarcinoma Cells A549 mediated by TGF-β1 was observed by MTT assay. Cellular immunofluorescence, western blot and real-time PCR was used to assay the protein and m RNA expression of MMP-9 and Integrinβ1. The data was checked with one factor analysis of variance using the SPSS17.0software, and ANOVA method was used to make comparison in the way of( X ± s), if P <0. 05, it would have significant difference.Results:1.Compared with A549 normal group, morphology changed in Cells A549 mediated by TGF-β1. A549 cells were observed by inverted phase contrast microscope to turn form pebble shape to fusiform shape, a myofibroblast-like morphology.2.Compared with blank serum group,high,middle and low dose groups of serum Buzhong Yiqi Decoction can improve the inhibition rate(IR) of Buzhong Yiqi Decoction on Intergrinβ1 of Human Lung Adenocarcinoma Cells A549 mediated by TGF-β1 significantly(P<0.01).3.12 h, high, middle dose groups of serum Buzhong Yiqi Decoction could inhibit cell migration by scratch experimental(P<0.05); 24 h, TGF-β1 induced group migrate faster than A549 normal group(P <0.01), high, middle and low dose groups of serum Buzhong Yiqi Decoction could inhibit cell migration by scratch experimental(P<0.01).4.Cellular immunofluorescence was used to assay the protein expression of MMP-9 and Integrinβ1. TGF-β1 induced group expressed strong positive than A549 normal group(P<0.01); Compared with TGF-β1 induced group, groups of serum Buzhong Yiqi Decoction could decreased the expression of MMP-9 and Integrinβ1(P<0.01).5. Western blot was used to assay the protein expression of MMP-9 and Integrinβ1.Compared with TGF-β1 induced group, groups of serum Buzhong Yiqi Decoction could decreased the expression of Integrinβ1(P<0.05); TGF-β1 induced group expressed strong positive than A549 normal group(P<0.05); high and middle dose groups of serum Buzhong Yiqi Decoction could decreased the expression of MMP-9(P<0.01).6. Real-time PCR was used to assay the m RNA expression of MMP-9 and Integrinβ1.TGF-β1 induced group expressed strong positive than A549 normal group(P<0.01);Compared with TGF-β1 induced group, groups of serum Buzhong Yiqi Decoction could decreased the m RNA expression of MMP-9 and Integrinβ1(P<0.01).Conclusion:1.Buzhong Yiqi Decoction Drug-containing serum suppress the growth of A549 obviously,and it is observed to exert a concentration-dependent.2. Buzhong Yiqi Decoction Drug-containing serum influence the mechanism of A549 EMT might be related to the down-regulation of MMP-9 and Integrinβ1 protein expression, and inhibite the tumor cell proliferation, Invasion and metastasisand.3.Buzhong Yiqi Decoction Drug-containing serum influence the mechanism of A549 EMT might be related to the down-regulation of MMP-9 and Integrinβ1 m RNA expression, and inhibite the tumor cell proliferation, Invasion and metastasisand. |
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