Fig. 2: LHPP restains metastatic ability of colorectal cancer cells in vivo and vitro. | Cell Death Discovery

Fig. 2: LHPP restains metastatic ability of colorectal cancer cells in vivo and vitro.

From: LHPP suppresses colorectal cancer cell migration and invasion in vitro and in vivo by inhibiting Smad3 phosphorylation in the TGF-β pathway

Fig. 2

Effect of LHPP overexpression on the migration and invasion (matrix gel 50–80 μl, dilution 1:8, 8-μm pore) of Sw480 (a) and Caco2 (b) cell lines compared to their corresponding control groups (scale bar, 100 μm, ×100). The quantification of migration and invasion in Sw480 cells (c) and Caco2 (d) were calculated using the Graphpad Prism 6 software according to Transwell assay, Migrated and invasive cells were stained and counted in at least five microscope fields. Blocking LHPP increased migration and invasion abilities of (matrix gel 50–80 μl, dilution 1:8, 8-μm pore) Sw480 (e) and HT-29 (f) cells when compared to negative groups (scale bar, 100 μm, ×100); The statistical differences were calculated by the Graphpad Prism 6 software HT-29 (g) and Sw480 (h). Migrated and invasive cells were stained and counted in at least five microscope fields. i Effect of LHPP overexpression on the migration of Sw480 cells at 24 and 48 h (scale bar 100 μm, ×100). j LHPP downregulation promoted HT-29 cells migration determined by wound-healing assay at 24 and 48 h (scale bar 100 μm, ×100). k The change of Caco2 cells’ shape following enhancement of LHPP expression; The change of HT-29 cells’ shape after reduction of LHPP expression (scale bar, 100 μm, ×100). l The representative images of lungs from nude mice (n = 5), black arrows showed tumor metastasis. m Lung nodules were identified using H&E staining. n Average number of tumor nodules in lung metastasis in Caco2 OE-LHPP group and their control group. o EMT-related proteins including E-cadherin, MMP2, Snail, Slug, and Twist1 were evaluated by using western blot in Sw480 cells after upregulating and downregulating LHPP expression. p Protein levels of E-cadherin, N-cadherin, MMP2, Snail, Slug, and Twist1 were tested using Western blot assay in Caco2 with high expression of LHPP and HT-29 with low expression of LHPP. q–t The Graphpad Prism 6 and Image J software were applied to observe the statistical differences between the experimental group and their control group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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