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Inhibitory Effects and Mechanism of Yiqi Chutan Fang on A549 Lung Cancer Metastasis in BALB/c-nu Nude Mice
Author(s): CHEN Changming, SUN Lingling, LIN Lizhu, Department of Oncology, the First Affiliated Hospital of Guangzhou University of Chinese Medicine
Pages: 513-
519
Year: 2016
Issue:
4
Journal: Traditional Chinese Drug Research & Clinical Pharmacology
Keyword: Yiqi Chutan Fang; A549 cells; Lung cancer metastasis; Epithelial-mesenchymal transition;
Abstract: Objective To observe the effect of Yiqi Chutan Fang(YCF)on inhibiting the tumor growth and metastasis of A549 lung cancer in BALB/c-nu nude mice model, and to preliminarily research the therapetuic mechanism.Methods A549 lung cancer xenograft model of nude mice was established. The model mice were randomly divided into blank control group and YCF group(15.5 g·kg-1·d-1),and were respectively given intragastric administration of saline and YCF for 21 continuous days. During treatment,we measured mice body weight and tumor volume every 3 days.After treatment for 21 days,all of the mice were sacrificed,and the tumor mass was stripped for measuring tumor weight and calculating tumor-inhibition rate;lung tissue was excised for counting lung surface metastatic nodules and calculating metastasis-suppression rate. We detected the expression of epithelial-mesenchymal transition(EMT)-related markers(E-cadherin,vimentin and fibronectin),GRP78,SRC,MAPK,PI3 K,AKT,smad2/3 and the phosphorylation protein in tumor tissue by immunohistochemistry. Results In YCF group,tumor volume and tumor weight were less than those of the blank control group(P < 0.05), and the tumor-inhibition rate of YCF group was 27.6 %,suggesting that YCF inhibit tumor growth. Lung metastatic nodules of mice in YCF group were significantly less than the blank control group(P < 0.05),and the tumor-metastasis inhibition rate was 39.6 %,suggesting that YCF significantly inhibit the metastasis of lung cancer. The results of immunohistochemistry showed that epithelial marker E-cadherin protein expression was increased(P < 0.05),whereas EMT-related markers vimentin and fibronectin were decreased significantly in YCF group(P < 0.05) compared with the blank control group, suggesting that YCF inhibit the occurrence of EMT,thereby reducing the invasion and metastasis of tumor cells. In YCF group,the phosphorylation and expression of GRP78,smad2/3,SRC,P38,ERK,and JNK were decreased(P < 0.05),suggesting that the anti-EMT mechanism of YCF may be related the multi-target suppression of signal proteins of GRP78, smad2/3,SRC, P38, ERK, and JNK. Conclusion YCF could inhibit the growth and metastasis of lung cancer, and the metastasis-suppression mechanism may be related with inhibiting the occurrence of EMT in cancer cells. YCF may be through inhibiting multi-target suppression of GRP78, smad2/3, SRC, MAPK signaling pathways to achieve the reverse effect on EMT.
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