Journal: Nature communications

Article Title: SCN4B acts as a metastasis-suppressor gene preventing hyperactivation of cell migration in breast cancer.

doi: 10.1038/ncomms13648

Figure Lengend Snippet: Figure 3 | Expression of the SCN4B/b4gene in human breast cancer cell lines and contribution to cancer cell invasiveness. (a) The expression of SCN4B gene was studied by RT–qPCR in human mammary epithelial non-cancer MCF-10A and cancer MCF7, MDA-MB-468, MDA-MB-435s and MDA-MB-231 cell lines. Results are expressed as relative to that of HPRT-1 gene (n ¼ 7–12) and presented as mean values±s.e.m. *, significantly different from MCF-10A at Po0.05 (MW). (b) The expression of SCN4B/b4 protein was studied by densitometric analysis of western blot experiments in same cells as in a. Results are given as the amount of SCN4B/b4 protein relative to that of HSC70 (n ¼ 5) and presented as mean values±s.e.m. *, significantly different from MCF-10A at Po0.05 (MW). The image on top shows a representative western blotting experiment. (c) The expression of SCNxB genes was analysed in MDA-MB-231-Luc cells by reverse transcription–PCR. Plasmids encoding human SCNxB genes were used as positive controls for PCR primers. (d) Representative western blotting experiments showing protein expression for b1 (SCN1B), b2 (SCN2B) and b4 (SCN4B) in MDA-MB-231-Luc cells. (e) Cells were transfected with scrambled siRNA (siCTL) or with siRNA directed against the expression of the SCN1B gene (siSCN1B), the SCN2B gene (siSCN2B) or the SCN4B gene (siSCN4B). The efficacy of siRNA transfection was assessed by western blotting experiments 48 h after transfection. HSC70 was used as a control for sample loading. (f) Representative images of fixed and haematoxylin-stained MDA-MB-231-Luc cells on invasion inserts. Cancer cells were transfected with scrambled siCTL or with specific siRNA. Scale bars, 50 mm. (g) Summary of cancer cell invasiveness results (n ¼ 8) for MDA-MB-231-Luc cells transfected with siCTL or siSCNxB. Results were expressed relative to siCTL and presented as mean values±s.e.m. ***, statistically different from siCTL at Po0.001 (Student’s t-test). (h) Representative image of a zebrafish embryo injected in the yolk sac with MDA-MB-231-Luc cells stained with CM-Dil and showing sites of colonization. Scale bars, 500 mm. Below is a magnification of the highlighted region containing human cancer cells (see arrows) colonizing organs of the embryo. (i) Zebrafish colonization index of siCTL or siSCN4B cells. Numbers in brackets indicate the number of embryos examined for each condition, from three different experiments. Results are presented as mean values±s.e.m. **, statistically different from siCTL at Po0.01 (Student’s t-test).

Article Snippet: MDA-MB-231-Luc human breast cancer cells were transfected with 20 nM siRNA targeting the expression of SCN1B (siSCN1B, sc-97849), SCN2B (siSCN2B, sc-96252), SCN4B (siSCN4B, sc-62982) or scrambled siRNA (siCTL, siRNA-A sc-37007), which were produced by Santa Cruz Biotechnology and were purchased from Tebu-Bio (France).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Reverse Transcription, Transfection, Control, Staining, Injection

Journal: International journal of oncology

Article Title: CDC20 associated with cancer metastasis and novel mushroom‑derived CDC20 inhibitors with antimetastatic activity.

doi: 10.3892/ijo.2019.4791

Figure Lengend Snippet: Figure 1. CDC20 knockdown inhibits the migration of chemoresistant pancreatic cancer and metastatic breast cancer cells. PANC‑1 cells and MDA‑MB‑231 cells were transfected with scrambled siRNA or CDC20 siRNA. (A) After 48 h of transfection, western blot analysis of CDC20 in PANC‑1 cells was evaluated. (B) Migration of PANC‑1 cells in Transwell chambers. (C) After 48 h of transfection, western blot analysis of CDC20 in MDA‑MB‑231 cells was evaluated. (D) Migration of MDA‑MB‑231 cells in Transwell chambers. Each bar represents the mean ± standard deviation of three individual filters within one repre sentative experiment repeated at least twice. **P<0.05 vs. Control. CDC20, cell division cycle 20; siRNA, small interfering RNA.

Article Snippet: The PANC-1 or MDA-MB-231 cells were transfected with human CDC20 siRNA (cat. no. sc-156154) or control siRNA-A (cat. no. sc-37007) using siRNA Transfection Reagent (cat. no. sc-29528) from Santa Cruz Biotechnology, Inc. as previously described (27).

Techniques: Knockdown, Migration, Transfection, Western Blot, Standard Deviation, Control, Small Interfering RNA

Journal: International journal of oncology

Article Title: CDC20 associated with cancer metastasis and novel mushroom‑derived CDC20 inhibitors with antimetastatic activity.

doi: 10.3892/ijo.2019.4791

Figure Lengend Snippet: Figure 2. Overexpression of CDC20 promotes the migration of chemoresistant pancreatic cancer and breast cancer. PANC‑1 cells and MCF‑7 cells were transfected with FLAG‑tagged plasmid DNA with CDC20 or FLAG‑tagged plasmid DNA. (A) After 24 h of transfection, western blot analysis of CDC20 in PANC‑1 cells was performed. (B) Migration of PANC‑1 cells in Transwell chambers. (C) After 24 h of transfection, western blot analysis of CDC20 in MCF‑7 cells was performed. (D) Migration of MCF‑7 cells in Transwell chambers was determined. Each bar represents the mean ± standard deviation of three individual filters within one representative experiment repeated at least twice. **P<0.05 and ***P<0.001 vs. Control. CDC20, cell division cycle 20.

Article Snippet: The PANC-1 or MDA-MB-231 cells were transfected with human CDC20 siRNA (cat. no. sc-156154) or control siRNA-A (cat. no. sc-37007) using siRNA Transfection Reagent (cat. no. sc-29528) from Santa Cruz Biotechnology, Inc. as previously described (27).

Techniques: Over Expression, Migration, Transfection, Plasmid Preparation, Western Blot, Standard Deviation, Control

Journal: International journal of oncology

Article Title: CDC20 associated with cancer metastasis and novel mushroom‑derived CDC20 inhibitors with antimetastatic activity.

doi: 10.3892/ijo.2019.4791

Figure Lengend Snippet: Figure 3. Effect of triterpenes from P. cocos on the expression of CDC20 in chemoresistant pancreatic cancer. PANC‑1 cells were treated with PTE (30 and 60 µg/ml), or PA (30 and 60 µM), DPA (30 and 60 µM) and PPAC (30 and 60 µM), three triterpenes purified from PTE for 24 h, respectively. Whole protein extracts isolated from cells were prepared and western blot analysis with anti‑CDC20 and anti‑β‑actin antibodies was performed. β‑actin was used as a loading control. Representative images are shown. Similar results were obtained in at least two additional experiments. P. cocos, Poria cocos; CDC20, cell division cycle 20; PTE, triterpene mixture extracted from P. cocos; PA, pachymic acid; DPA, purified triterpenes dehydropachymic acid; PPAC, polyporenic acid C.

Article Snippet: The PANC-1 or MDA-MB-231 cells were transfected with human CDC20 siRNA (cat. no. sc-156154) or control siRNA-A (cat. no. sc-37007) using siRNA Transfection Reagent (cat. no. sc-29528) from Santa Cruz Biotechnology, Inc. as previously described (27).

Techniques: Expressing, Purification, Isolation, Western Blot, Control