Journal: International Journal of Oncology

Article Title: Loss of profilin 2 contributes to enhanced epithelial-mesenchymal transition and metastasis of colorectal cancer

doi: 10.3892/ijo.2018.4475

Figure Lengend Snippet: PFN2 expression level is associated with EMT-related molecules and migratory activity in CRC cells. (A) The expression of EMT-related molecules in human normal colorectal, non-metastatic CRC and metastatic CRC tissues. (B and C) mRNA and protein expression levels of PFN2 and EMT related molecules, E-cadherin, N-cadherin, vimentin, slug and snail, were measured in two CRC cell lines, HCT116 and SW620, by (B) reverse transcription-quantitative polymerase chain reaction and (C) western blotting, respectively; GAPDH served as the internal reference. (D and E) The migratory activity of CRC cell lines was analyzed by (D) wound-healing and (E) Transwell migration assays at 24 h. Data are presented as the mean ± sandard error of the mean of three independent experiments. * P<0.05, ** P<0.01 and *** P<0.001. CRC, colorectal cancer; EMT, epithelial-mesenchymal transition; N, normal; N-M, non-metastatic; M, metastatic; PFN2, profilin 2.

Article Snippet: To establish the liver cancer metastasis model, mice were anesthetized, a transverse incision was made in the left flank to expose the spleen and 1×10 6 PFN2-OE-transfected or empty vector-transfected SW620 cells in 100 µ l phosphate-buffered saline (PBS; Thermo Fisher Scientific, Inc.) were injected intrasplenically (n=5 mice/group).

Techniques: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Migration

Journal: International Journal of Oncology

Article Title: Loss of profilin 2 contributes to enhanced epithelial-mesenchymal transition and metastasis of colorectal cancer

doi: 10.3892/ijo.2018.4475

Figure Lengend Snippet: PFN2 overexpression results in decreased EMT in colorectal cancer. PFN2 was transiently overexpressed in SW620 cells using a pQCXIH-PFN2 vector. (A) Western blotting was performed to confirm the overexpression efficiency in PFN2-OE transfected SW620 cells; untransfected and empty vector-transfected cells were used as controls. (B) Protein expression levels of EMT markers and regulators were analyzed by western blotting; GAPDH served as the internal reference. (C and D) Migratory abilities of the transfected SW620 cells were examined by (C) wound-healing and (D) Transwell migration assays, respectively. (E and F) Vector-transfected control SW620 cells or PFN2-OE-transfectd SW620 cells were injected into nude mice (E) via the spleen to induce liver metastasis or (F) via the tail vein to induce lung metastasis. The yellow arrows indicate the metastatic tumor nodules and hematoxylin and eosin staining was performed to confirm the tumor characteristics. (G) The volumes of the largest metastatic tumor nodules were calculated. Data are presented as the mean ± sandard error of the mean of three independent experiments. * P<0.05 and ** P<0.01. Ctrl, control; EMT, epithelial-mesenchymal transition; OE, overexpression; PFN2, profilin 2.

Article Snippet: To establish the liver cancer metastasis model, mice were anesthetized, a transverse incision was made in the left flank to expose the spleen and 1×10 6 PFN2-OE-transfected or empty vector-transfected SW620 cells in 100 µ l phosphate-buffered saline (PBS; Thermo Fisher Scientific, Inc.) were injected intrasplenically (n=5 mice/group).

Techniques: Over Expression, Plasmid Preparation, Western Blot, Transfection, Expressing, Migration, Injection, Staining

Journal: International Journal of Oncology

Article Title: Loss of profilin 2 contributes to enhanced epithelial-mesenchymal transition and metastasis of colorectal cancer

doi: 10.3892/ijo.2018.4475

Figure Lengend Snippet: Cytoskeletal remodeling may be involved in PFN2-regulated EMT. (A and B) p-MLC expression was measured by western blotting in (A) normal colon, non-metastatic CRC and metastatic CRC tissues, and (B) in untransfected, vector- and PFN2-OE-transfected SW620 cells. (C) Protein expression levels of PFN2, p-MLC, E-cadherin and vimentin were examined by western blotting in PFN2-OE-transfected SW620 cells with or without co-treatment with Y27632; β-tubulin served as the internal reference. (D) Wound-healing assays were performed to determine the migratory ability of PFN2-OE-transfected SW620 with or without co-treatment with Y27632. Data are presented as the mean ± sandard error of the mean of three independent experiments. * P<0.05, ** P<0.01 and *** P<0.001. CRC, colorectal cancer; EMT, epithelial-mesenchymal transition; MLC, myosin light chain; OE, overexpression; p, phosphorylated; PFN2, profilin 2; Y, Rho-associated kinase inhibitor Y27632.

Article Snippet: To establish the liver cancer metastasis model, mice were anesthetized, a transverse incision was made in the left flank to expose the spleen and 1×10 6 PFN2-OE-transfected or empty vector-transfected SW620 cells in 100 µ l phosphate-buffered saline (PBS; Thermo Fisher Scientific, Inc.) were injected intrasplenically (n=5 mice/group).

Techniques: Expressing, Western Blot, Plasmid Preparation, Transfection, Over Expression