Figure 1.
FoxM1 overexpression correlated with substantially poor prognosis of NSCLC patients.
(A) Representative results of FoxM1 staining in NSCLC tumor tissues analyzed by immunohistochemistry (a, negative; b, weak; c, moderate; d, intense, respectively). Representative results of FoxM1 expression levels between lymph node positive (e) and lymph node negative (f) (a-f, left, magnification: ×100; right, magnification: ×400). (B) Overall survival analyses for all patients (a), patients with stage I/II (b) and patients with stage III/IV (c) according to the results of immunohistochemistry analysis. The log-rank test was used to test the two survival distributions. P<0.05 was considered to have statistical significance.
Table 1.
Univariate Survival Analysis (n = 175).
Table 2.
Multivariable analysis for the effect of FoxM1 expression on survival.
Figure 2.
Targeted knockdown of FoxM1 expression inhibited the in vitro metastatic potentials of NSCLC cells.
(A, B) The detection of lentivirus-mediated knockdown of FoxM1 in PC-9 and H1299 cells by WB and q-PCR, respectively. (C, D) A representative result of the trans-well migration assays for the effects of FoxM1 on the in vitro migratory and invasive abilities of PC-9 and H1299 cells. The results are shown as the mean ± s.d., *p<0.05.
Figure 3.
Ectopic expression of FoxM1 increased the in vitro metastatic potentials of NSCLC cells.
(A, B) The detection of lentivirus-mediated overexpression of FoxM1 in PC-9 and H292 cells by WB and q-PCR, respectively. (C, D) A representative result of the trans-well migration assays for the effects of FoxM1 on the in vitro migratory and invasive abilities of PC-9 and H292 cells. The results are shown as the mean ± s.d., *p<0.05.
Figure 4.
Enforced expression of FoxM1 promoted the in vivo metastatic abilities of NSCLC cells.
(A, B) Visual inspection and fluorescence imaging analysis of lung metastatic nodules in two groups (PC-9-Vector and PC-9-FoxM1) of mouse models by tail vein injection of tumor cells, respectively. (C) The effects of FoxM1 on the in vivo metastatic abilities of PC-9 cells in xenograft models of nude mice (n = 6) as determined by examination of mouse lungs for microscopic nodules. Representative results of histological examination of mouse lungs for metastatic nodules (left). The number of lung metastatic noudels in two groups of mouse models (right). The results are shown as the mean ± s.d., *p<0.05.
Figure 5.
FoxM1 overexpression induced the epithelial-mesenchymal transition of NSCLC cells.
(A) Representative phase-contrast images of the H292-vector and H292-FoxM1 cells (Magnification: 200×). (B) quantitive real-time PCR analysis of the relative expression of EMT markers as indicated. GADPH were used as loading control. Data are expressed as Mean ±s.d., *p<0.05. (C) The relative expression of EMT markers as indicated was determined by immunoblotting (left), and the results of which was further quantified (right). β-actin served as a loading control. (D) The relative expression of Slug as indicated was determined by immunoblotting (left), and the results of which was further quantified (right). β-actin served as a loading control.
Figure 6.
FoxM1 overexpression induced the epithelial-mesenchymal transition of NSCLC tissue samples.
(A) The relative expression of E-cadherin, vimentin and Slug as indicated was determined by immunohistochemical staining in human tumor samples with high (above) and low (below) FoxM1 expression. (B) The relative expression of E-cadherin, vimentin and Slug as indicated was determined by immunohistochemical staining in nude mice tumors derived from PC-9-vector cells (above) and PC-9-FoxM1 cells (below).