Ter, Third Affiliated Crizotinib Hospital of Kunming Medical University (Tumor Hospital of Yunnan Province), Kunming, Yunnan, People's Republic China.According to our previous study [17], a total of 1 ?106 logarithmically growing A549 cells transfected with fulllength ENO1 and PLV-Ctr vector, SPCA-1 cells transfected with shENO1-B, and the control PLV-shCtr vector (N = 6 per group) in 0.1 ml Hank's solution were subcutaneously inoculated into the left-right symmetric flank of 4?-week-old male BALB/c-nu/nu mice. The mice were maintained in a barrier facility on HEPA-filtered racks and fed an autoclaved laboratory rodent diet. All animal studies were conducted in accordance with the principles and procedures outlined in the National Institutes of Health Guide for the Care and Use of Animals under assurance number A3873-1. After 15 days, the mice were sacrificed, and their tumors were excised, weighed, and processed for histology.In vivo metastasis assaysReceived: 15 October 2014 Accepted: 9 FebruaryIn vivo metastasis assays were performed according to a previous study [46]. A total of 5 ?106 cells were injected into nude mice (n = 5 for each group) through the spleen, respectively. The optical fluorescence images were visualized to monitor primary tumor growth and formation of metastatic lesions. Forty days later, all mice were killed, individual organs were removed, and metastatic tissues were analyzed by H E staining.References 1. Yuan X, Wu H, Han N, Xu H, Chu Q, Yu S, et al. Notch signaling and EMT in non-small cell lung cancer: biological significance and therapeutic application. J Hematol Oncol. 2014;7(1):87. 2. Molina JR, Yang P, Cassivi SD, Schild SE, Adjei AA. Non-small cell lung cancer: epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc. 2008;83(5):584?4. 3. Gomez-Casal R, Bhattacharya C, Ganesh N, Bailey L, Basse P, Gibson M, et al. Non-small cell lung cancer cells survived ionizing radiation treatment display cancer stem cell and epithelial-mesenchymal transition phenotypes. Mol Cancer. 2013;12(1):94.Fu et al. Journal of Hematology Oncology (2015)8:Page 13 of4.5.6. 7.8.9.10.11.12. 13.14.15.16.17.18.19.20.21.22.23.24.25.26.27.Singh S, Trevino J, Bora-Singhal N, Coppola D, Haura E, Altiok S, et al. EGFR/ Src/Akt signaling modulates Sox2 expression and self-renewal of stem-like side-population cells in non-small cell lung cancer. Mol Cancer. 2012;11:73. Serrano D, Bleau AM, Fernandez-Garcia I, Fernandez-Marcelo T, Iniesta P, Ortiz-de-Solorzano C, et al. Inhibition of telomerase activity preferentially targets aldehyde dehydrogenase-positive cancer stem-like cells in lung cancer. Mol Cancer. 2011;10:96. Jin S, DiPaola RS, Mathew R, White E. Metabolic catastrophe as a means to cancer cell death. J Cell Sci. 2007;120(Pt 3):379?3. Diaz-Ramos A, Roig-Borrellas A, Garcia-Melero A, Lopez-Alemany R. Alpha-enolase, a multifunctional protein: its role on pathophysiological situations. J Biomed Biotechnol. 2012;2012:156795. Merkulova T, Dehaupas M, Nevers MC, Creminon C, Alameddine H, Keller A. Differential modulation of alpha, beta and gamma enolase isoforms in regenerating mouse skeletal muscle. Eur J Biochem. 2000;267(12):3735?3. Feo S, Arcuri D, Piddini E, Passantino R, Giallongo A. ENO1 gene product binds to the c-Myc promoter and acts as a transcriptional repressor: relationship with Myc promoter-binding protein 1 (MBP-1). Febs Lett. PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/7500280 2000;473(1):47?2. Lung J, Liu KJ, Chang JY, Leu SJ, Shih NY. MBP-1 is efficiently encoded.