Journal: Molecular Oncology

Article Title: JNK‐mediated Ser27 phosphorylation and stabilization of SIRT1 promote growth and progression of colon cancer through deacetylation‐dependent activation of Snail

doi: 10.1002/1878-0261.13143

Figure Lengend Snippet: Overexpression of P‐SIRT1 Ser27 in CRC tissues and its implications for SIRT1 stability and proliferation and migration of colon cancer cells. (A) SW480 cells (5 × 10 6 ) stably expressing mock or SIRT1 vector were injected subcutaneously into the flank of BALB/c nude mice. Photographs show the xenograft tumors resected from mice at the time of harvest (day 50). The tumor volume and the tumor weight of the mice at the time of harvest were measured. Comparison of the means between mock and SIRT1 vector group was determined by Student's t ‐test. The results are shown as the mean ± S.D. of four xenografts for each group: * P < 0.05. Scale bar represents 1 cm. (B) Histological structures in sections from excised xenograft tumors were analyzed by H&E staining (first column). The yellow arrow indicates neovascularization. The expression of SIRT1, IL‐6, and 8 in excised xenograft tumors were examined by immunohistochemical analysis (second to last columns). Each scale bar represents 100 µm. The data are presented as the mean ± S.D. of three independent samples for each group in Fig. S1H: * P < 0.05; ** P < 0.01. (C) Kaplan–Meier survival curve of overall survival in colon cancer patients with high ( n = 497) and low ( n = 506) levels of SIRT1 based on the data collected from the GENT2 database ( http://gent2.appex.kr ). (D) Kaplan–Meier survival curve of disease‐specific survival in colon cancer patients with high ( n = 238) and low ( n = 238) levels of SIRT1 based on the data obtained from the GENT2 database ( http://gent2.appex.kr ) (E) Immunofluorescence staining of tissue microarray containing thirty‐five pairs of colon adenocarcinoma and matched adjacent normal colon tissues was conducted using anti‐SIRT1 and P‐SIRT1 Ser27 antibodies. The adjacent normal area is 1.5 cm distant from the tumor, which was taken by a histologist. Images of H&E‐stained tissue sections were offered by US Biomax Inc. Each scale bar represents 200 µm. (F) The fluorescence intensity was measured by image analysis software ImageJ, and results are shown as a violin plot. Statistics were calculated using both paired t ‐test and Wilcoxon matched‐pairs test. (G) The protein levels of SIRT1 and P‐SIRT1 Ser27 in colon cancer and corresponding normal tissues were measured by Western blot analysis. The data are presented as the mean ± S.D. of eleven pairs of human tissue specimens in Fig. D: * P < 0.05.

Article Snippet: Two human paraffin‐embedded colon cancer tissue array slides with matched adjacent normal colon tissue (US Biomax, Inc., cat. no. CO703; Rockville, MD, USA) were subjected to deparaffinization with xylene.

Techniques: Over Expression, Migration, Stable Transfection, Expressing, Plasmid Preparation, Injection, Staining, Immunohistochemical staining, Immunofluorescence, Microarray, Fluorescence, Software, Western Blot

Journal: bioRxiv

Article Title: TMIGD1, a putative tumor suppressor, induces G2-M cell cycle checkpoint arrest in colon cancer cells

doi: 10.1101/2020.06.06.138057

Figure Lengend Snippet: ( A ) Two human colorectal cancer tissue microarray slides consist of 172 tissue samples were stained with anti-TMIGD1 antibody. Staining was scored as 0 (negative, <5% cells positive), 1+ (6-25% cells positive), 2+ (26-50% cells positive), and 3+ (>50% cells positive). The tumor differentiation was graded morphologically (grade 1, 2 and 3 as well, moderately and poorly differentiated). ( B ) The average staining intensities were compared using ANOVA with Tukey post-hoc test per tumor grade. ( C ) The average staining intensities were compared using ANOVA with Tukey post-hoc test per tumor differentiation. ( D ) Western blot analysis of TMIGD1 expression in human renal and colorectal cancer cell lines. ( E ) qPCR analysis of TMIGD1 expression in CRC cell lines (HT29, HCT116 and RKO). Human kidney epithelial cells, HK2 was used as a positive control.

Article Snippet: Two human colorectal cancer tissue microarray slides (US Biomax, catalog numbers BC05012a and BC05118a) consist of 172 tissue samples (72 on BC05012a and 100 on BC05118a) were stained with anti-TMIGD1 antibody.

Techniques: Microarray, Staining, Western Blot, Expressing, Positive Control