human crc cell line colo201  (JCRB Cell Bank)

Journal: Translational and Regulatory Sciences

Article Title: Suppression of tensin 2 promotes intestinal tumorigenesis by liberating integrin-linked kinase-induced nuclear translocation of β-catenin

doi: 10.33611/trs.2020-009

Figure Lengend Snippet: Fig. 1. Tns2 suppresses the proliferation and migration of colorectal cancer cell lines. (A) RT-qPCR analysis showed that TNS2 mRNA expression level was lower in HCT116 cells than in Colo201 and SW620 cells. In Colo201 and SW620 cells, siRNA treatment sup- pressed TNS2 mRNA expression. Relative fold changes in TNS2 expression were calculated using the comparative 2−ΔΔCT method and normalized to TNS2 level from HCT116 cells. (B) RT-PCR analysis showed the stable expression of murine Tns2 in HCT116 cells. Minus or plus indicates the absence or presence of pCAG-Tns2, respectively. (C–E) Proliferation of colorectal cancer cell lines in the presence or absence of TNS2 gene. TNS2 KD significantly increased the proliferation of Colo201 (C) and SW620 cells (D) (n=3). Tns2 overexpression induced a significant decrease in HCT116 cell proliferation (E) (n=4). (F–H) Migration of colorectal cancer cell lines in the presence or absence of TNS2 gene. TNS2 KD significantly increased the migration of Colo201 (F) and SW620 (G) cells (n=4). Tns2 overexpression induced a significant decrease in HCT116 cell migration (H) (n=4). Values are means ± SD. *P<0.05, ** P<0.01, ***P<0.001.

Article Snippet: Human CRC cell lines Colo201, SW620, and HCT116 were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Nacalai Tesque, Kyoto, Japan), Leibovitz’s L-15 medium (Wako, Osaka Japan), and Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque) supplemented with 10% fetal bovine serum (HyClone, Logan, UT, USA), 100 units/ml of penicillin, and 100 μg/ml of streptomycin, respectively.

Techniques: Migration, Quantitative RT-PCR, Expressing, Reverse Transcription Polymerase Chain Reaction, Over Expression

Journal: Translational and Regulatory Sciences

Article Title: Suppression of tensin 2 promotes intestinal tumorigenesis by liberating integrin-linked kinase-induced nuclear translocation of β-catenin

doi: 10.33611/trs.2020-009

Figure Lengend Snippet: Fig. 2. Tns2 suppresses the Wnt/β-catenin activity and nuclear translocation of β-catenin. (A)TNS2 KD increases Wnt/β-catenin activity in Colo201 cells, as detected by the TOPFlash luciferase assay (n=6). (B) Tns2 overexpression decreases Wnt/β-catenin activity in HCT116 cells, as detected by the TOPFlash luciferase assay (n=6). (C) β-catenin level was similar between the whole cell lysates of HCT116-Tns2 cells and control cells. (D) Tns2 overexpression decreased the nuclear translocation of β-catenin in HCT116 cells, as detected by immunofluorescence (Control: n=221, Tns2: n=117). White arrows indicate the position of the nucleus. Values are means ± SD. *P<0.05, **P<0.01.

Article Snippet: Human CRC cell lines Colo201, SW620, and HCT116 were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Nacalai Tesque, Kyoto, Japan), Leibovitz’s L-15 medium (Wako, Osaka Japan), and Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque) supplemented with 10% fetal bovine serum (HyClone, Logan, UT, USA), 100 units/ml of penicillin, and 100 μg/ml of streptomycin, respectively.

Techniques: Activity Assay, Translocation Assay, Luciferase, Over Expression, Control, Immunofluorescence