Nitidine chloride suppresses epithelial-to-mesenchymal transition in osteosarcoma cell migration and invasion through Akt/GSK-3β/Snail signaling pathway.
Metastasis is the main cause of death in osteosarcoma. Targeting the process of metastasis is a main strategy for osteosarcoma therapy. As a traditional Chinese medicine, Zanthoxylum nitidum (Roxb) has been applied to treat various diseases, including cancer. However, no evidence has been shown on the anti-metastasis effect of nitidine chloride (NC) that was extracted from Zanthoxylum nitidum (Roxb) on osteosarcoma cells, or its underling mechanisms. In the present study, we aimed to demonstrate the role of NC on the migration and invasion of osteosarcoma cells. Viability and proliferation of osteosarcoma cells were examined by MTT assay. Then, by appling scratch wound healing assay and Transwell assays, we evaluated migratory and invasive ability of the cells, respectively. Moreover, the expression of epithelial-to-mesenchymal transition (EMT) markers were determined after treatment with NC. Furthermore, the expression of Akt, GSK-3β and Snail were detected by western blot analysis. In addition, the GSK-3β activity was examined by GSK-3β kinase assay. Finally, an inhibitor of GSK-3β, lithium chloride (LiCl) was applied to testify the effect of NC on the expression of EMT markers and Snail. We found that the proliferative, migratory and invasive ability of the U2OS osteosarcoma cells were all suppressed when treated with NC. NC increased the expression of E-cadherin and decreased the expression of N-cadherin, vimentin and fibronectin in a dose-dependent manner. NC also exerted its ability to suppress the phosphorylation of Akt and GSK-3β so as to activate GSK-3β. Then, by using an GSK-3β inhibitor, LiCl, we revealed the effect of GSK-3β in the expression of EMT markers. The expression of Snail was inhibited when treated with NC and LiCl also reversed the NC-inhibited Snail expression. Taken together, these results revealed that NC suppressed EMT and decreased the invasive ability of osteosarcoma cells via the Akt/GSK-3β/snail signaling pathway.
Department of Neurosurgery, People's Hospital of Linyi City, Linyi, Shandong, P.R. China.
Zhenxiu
Z
Cheng
Department of Osteology, Affiliated Hospital of Shandong Medical College, Linyi, Shandong, P.R. China.
Yinglong
Y
Guo
Department of Joint Surgery, Affiliated Hospital of Shandong Medical College, Linyi, Shandong, P.R. China.
Yubao
Y
Yang
Department of Joint Surgery, People's Hospital of Linyi City, Linyi, Shandong, P.R. China.
Jinqing
J
Kan
Department of Osteology, Qingdao Shili Hospital of Eastern Hospital, Qingdao, Shandong, P.R. China.
Shiyou
S
Dai
Department of Pathology, Central Hospital of Zibo, Zibo, Shandong, P.R. China.
Mengfei
M
Helian
Department of Cardiology, Central Hospital of Zibo, Zibo, Shandong, P.R. China.
Bo
B
Li
Genetic Disease Diagnosis Center, Zibo Maternal and Child Health Hospital, Affiliated to Shandong Academy of Medical Science, Zibo, Shandong, P.R. China.
Jia
J
Xu
Department of Osteology, People's Hospital of Linyi City, Linyi, Shandong, P.R. China.
Changying
C
Liu
D053119:Benzophenanthridines / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000235:genetics; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D015820:Cadherins / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D005353:Fibronectins / Q000235:genetics; D000071679:Glycogen Synthase Kinase 3 beta / Q000235:genetics*; D006801:Humans; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000473:pathology; D012516:Osteosarcoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects / Q000235:genetics; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D000071250:Snail Family Transcription Factors / Q000235:genetics*; D014746:Vimentin / Q000235:genetics
D053119:Benzophenanthridines; D014408:Biomarkers, Tumor; D015820:Cadherins; D005353:Fibronectins; D000071250:Snail Family Transcription Factors; D014746:Vimentin; C013615:nitidine; D000071679:Glycogen Synthase Kinase 3 beta; D051057:Proto-Oncogene Proteins c-akt