anti-human mouse monoclonal antibodies against cyclin d1  (Santa Cruz Biotechnology)

Journal: Oncology reports

Article Title: Downregulation of miR-34a contributes to the proliferation and migration of laryngeal carcinoma cells by targeting cyclin D1.

doi: 10.3892/or.2016.4823

Figure Lengend Snippet: Figure 3. CCND1 is a potential target of miR-34a. (A) Schematic of the putative miR-34a binding site in the 3'-UTR region of CCND1 and interspecies conservation of seed matching sequences (gray box). (B) Diagram of CCND1 3'-UTR containing the reporter constructs. (C) Luciferase reporter assays in HEp-2 cells co-transfected with wt/mut 3'-UTR and miR-34a/miR-Ctrl as indicated. (D) Expression levels of CCND1 were tested after miR-34a transfection at 50 nM in HEp-2 cells by western blotting assay. *P<0.05 compared with the control. 3'-UTR, 3'-untranslated region.

Article Snippet: After blocking for 1 h at room temperature and incubation overnight at 4̊C in Tris-buffered saline containing 0.05% Tween (TbsT) with 5% milk, the membranes were incubated with mouse monoclonal antibody against human CCND1 (Cell Signaling Technology, Danvers, MA, USA) or tubulin (Sigma-Aldrich) as a protein loading control, followed by horseradish peroxidase (HRp)-conjugated goat-anti-mouse igg (Abcam), and the bands were detected using the Supersignal West Pico eCL chemiluminescence kit (pierce) and Kodak X-ray film (Eastman Kodak Co, Rochester, NY, USA).

Techniques: Binding Assay, Construct, Luciferase, Transfection, Expressing, Western Blot, Control

Journal: Oncology reports

Article Title: Downregulation of miR-34a contributes to the proliferation and migration of laryngeal carcinoma cells by targeting cyclin D1.

doi: 10.3892/or.2016.4823

Figure Lengend Snippet: Figure 4. CCND1 is essential for LSCC proliferation and migration, and is involved in the miR-34a-induced effect. (A) Analysis of CCND1 protein expres- sion by western blot analysis. CCND1 protein was reduced by small interfering RNA (CCND1-siRNA) in the HEp-2 cells. (B) MTT assay showed that cell proliferation was inhibited in the HEp-2 cells after transfection with CCND1-siRNA, compared with the scrambled sequence (Ctrl-siRNA). (C) Apoptosis analysis of HEp-2 cells in response to CCND1-siRNA at 48 h, by FACS assay. (D) Analysis of the effect of CCND1-siRNA on the migration of HEp-2 cells by Transwell migration assay. (E) MTT assay shows the cell proliferation in HEp-2 cells co-transfected with miR-34a and wt/mut 3'-UTR-CCND1 compared with miR-Ctrl. *P<0.05 compared with the control. LSCC, laryngeal squamous cell carcinomas.

Article Snippet: After blocking for 1 h at room temperature and incubation overnight at 4̊C in Tris-buffered saline containing 0.05% Tween (TbsT) with 5% milk, the membranes were incubated with mouse monoclonal antibody against human CCND1 (Cell Signaling Technology, Danvers, MA, USA) or tubulin (Sigma-Aldrich) as a protein loading control, followed by horseradish peroxidase (HRp)-conjugated goat-anti-mouse igg (Abcam), and the bands were detected using the Supersignal West Pico eCL chemiluminescence kit (pierce) and Kodak X-ray film (Eastman Kodak Co, Rochester, NY, USA).

Techniques: Migration, Western Blot, Small Interfering RNA, MTT Assay, Transfection, Sequencing, Transwell Migration Assay, Control

Journal: Oncology reports

Article Title: Downregulation of miR-34a contributes to the proliferation and migration of laryngeal carcinoma cells by targeting cyclin D1.

doi: 10.3892/or.2016.4823

Figure Lengend Snippet: Figure 5. miR-34a and CCND1 are inversely correlated in LSCC tissues. (A) Statistical quantification of the mRNA expression levels of CCND1 (analyzed with q-PCR) between LSCC and paired NAT specimen. The CCND1 level was normalized to GAPDH (P<0.01). (B) A scatter diagram shows an inverse correlation between miR-34a and CCND1 expression in the same set of LSCC tissues (Spearman's correlation analysis, r =-7604; P<0.0001). NATs, normal adjacent tissues; LSCC, laryngeal squamous cell carcinomas.

Article Snippet: After blocking for 1 h at room temperature and incubation overnight at 4̊C in Tris-buffered saline containing 0.05% Tween (TbsT) with 5% milk, the membranes were incubated with mouse monoclonal antibody against human CCND1 (Cell Signaling Technology, Danvers, MA, USA) or tubulin (Sigma-Aldrich) as a protein loading control, followed by horseradish peroxidase (HRp)-conjugated goat-anti-mouse igg (Abcam), and the bands were detected using the Supersignal West Pico eCL chemiluminescence kit (pierce) and Kodak X-ray film (Eastman Kodak Co, Rochester, NY, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Concomitant depletion of PTEN and p27 and overexpression of cyclin D1 may predict a worse prognosis for patients with post-operative stage II and III colorectal cancer

doi: 10.3892/ol.2014.2350

Figure Lengend Snippet: Immunohistochemistry for PTEN, p27 and Cyclin D1 in CRC tissues and normal colonic mucosa adjacent to cancer tissues. Representative slides demonstrating immunohistochemical staining of CRC and adjacent normal mucosa. (A) Expression of PTEN is strongly detected in the cytoplasm of the CRC cells, but not in the nucleus. (B) PTEN expression is also observed in the matched normal colonic tissues. (C) The tumor cells exhibit diffuse nuclear staining for p27. (D) Strong nuclear staining is present in the matched normal colonic tissues. (E) Cyclin D1 immunoreactivity is confined to the nucleus in the CRC tissues. (F) No staining is present in the normal matched colonic tissues. All images are captured at ×40 magnification. PTEN, phosphatase and tensin homolog; CRC, colorectal cancer.

Article Snippet: Subsequent to blocking non-specific binding with 10% bovine serum, the sections were incubated with anti-human mouse monoclonal antibodies against PTEN (1:100), p27 (1:100) and cyclin D1 (1:100) (all Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C.

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining, Expressing

Journal: Oncology Letters

Article Title: Concomitant depletion of PTEN and p27 and overexpression of cyclin D1 may predict a worse prognosis for patients with post-operative stage II and III colorectal cancer

doi: 10.3892/ol.2014.2350

Figure Lengend Snippet: PTEN, p27 and Cyclin D1 expression in the control and CRC groups.

Article Snippet: Subsequent to blocking non-specific binding with 10% bovine serum, the sections were incubated with anti-human mouse monoclonal antibodies against PTEN (1:100), p27 (1:100) and cyclin D1 (1:100) (all Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C.

Techniques: Expressing, Control

Journal: Oncology Letters

Article Title: Concomitant depletion of PTEN and p27 and overexpression of cyclin D1 may predict a worse prognosis for patients with post-operative stage II and III colorectal cancer

doi: 10.3892/ol.2014.2350

Figure Lengend Snippet: Correlation between expression of PTEN, p27 and Cyclin D1 and clinicopathological characteristics in 61 patients with CRC.

Article Snippet: Subsequent to blocking non-specific binding with 10% bovine serum, the sections were incubated with anti-human mouse monoclonal antibodies against PTEN (1:100), p27 (1:100) and cyclin D1 (1:100) (all Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Concomitant depletion of PTEN and p27 and overexpression of cyclin D1 may predict a worse prognosis for patients with post-operative stage II and III colorectal cancer

doi: 10.3892/ol.2014.2350

Figure Lengend Snippet: Overall survival curves for patients with CRC based on the level of PTEN, p27 and Cyclin D1 expression. (A) The overall survival time for the patients with PTEN-positive expression was higher than that for the patients with PTEN depletion (χ 2 =28.71; P<0.001). (B) The patients who expressed high levels of p27 survived longer than those who demonstrated p27 depletion (χ 2 =26.88; P<0.001). (C) The patients with Cyclin D1-positive expression exhibited shorter survival times compared with those with no staining for Cyclin D1 (χ 2 =5.36; P=0.021). Analyses were performed using the Kaplan-Meier method. PTEN, phosphatase and tensin homolog; CRC, colorectal cancer.

Article Snippet: Subsequent to blocking non-specific binding with 10% bovine serum, the sections were incubated with anti-human mouse monoclonal antibodies against PTEN (1:100), p27 (1:100) and cyclin D1 (1:100) (all Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C.

Techniques: Expressing, Staining

Journal: Oncology Letters

Article Title: Concomitant depletion of PTEN and p27 and overexpression of cyclin D1 may predict a worse prognosis for patients with post-operative stage II and III colorectal cancer

doi: 10.3892/ol.2014.2350

Figure Lengend Snippet: Overall survival curves for CRC patients based on combined detection of PTEN/p27, PTEN/Cyclin D1 and p27/Cyclin D1 expression. (A) Patients with PTEN(+), PTEN(−), p27(+), p27(−), PTEN(+)/p27(+), and PTEN(−)/p27(−) exhibited a mean survival time of 77.1, 51.5, 74.7, 48.7, 77.1 and 48.7 months, respectively. Patients with PTEN(−)/p27(−) expression exhibited the worst survival times. (B) Patients with PTEN(+), PTEN(−), Cyclin D1(−), Cyclin D1(+), PTEN(+)/Cyclin D1(−) and PTEN(−)/Cyclin D1(+) expression had a mean survival time of 77.1, 51.5, 73.8, 56.9, 76.8 and 46.6 months, respectively. Patients with PTEN(+) expression exhibited the best survival times, while joint examination of PTEN(+)/Cyclin D1(−) expression did not provide extra prognostic information. However, combinations of PTEN(−)/Cyclin D1(+) expression provided a more adverse prognosis compared with detection of PTEN(−) and Cyclin D1(+) expression alone. (C) Patients with p27(+), p27(−), Cyclin D1(−), Cyclin D1(+), p27(+)/Cyclin D1(−) and p27(−)/Cyclin D1(+) expression had a mean survival time of 74.7, 48.7, 73.8, 56.9, 74.2 and 43.5 months, respectively. Combination of p27(+)/Cyclin D1(−) expression did not provide additional predictive information for clinical outcome compared with detection of p27(+) and Cyclin D1(−) expression alone. Notably, patients with p27(−)/Cyclin D1(+) expression had worse survival times. PTEN, phosphatase and tensin homolog; CRC, colorectal cancer.

Article Snippet: Subsequent to blocking non-specific binding with 10% bovine serum, the sections were incubated with anti-human mouse monoclonal antibodies against PTEN (1:100), p27 (1:100) and cyclin D1 (1:100) (all Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C.

Techniques: Expressing