Journal: Frontiers in Oncology

Article Title: Downregulated ARID1A by miR-185 Is Associated With Poor Prognosis and Adverse Outcomes in Colon Adenocarcinoma

doi: 10.3389/fonc.2021.679334

Figure Lengend Snippet: Differential expression of ARID1A. (A) Differential ARID1A expression analysis across several cancer types (significant differences by the Mann-Whitney U test are marked with red*). Left, normal; right, tumor); (B) ARID1A expression include paired tumor and adjacent normal tissues from Gene chip data; (C) ARID1A expression include paired tumor and adjacent normal tissues from RNA-Seq data; (D) Significant different expression of ARID1A through metastases, normal, and tumors; (E–F) Experimental verification of ARID1A mRNA and protein levels in colon cancer cell line using qPCR and western blot, respectively.

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline (1× TBS) containing 0.05% Tween-20, the membrane was incubated overnight at 4°C with rabbit antibodies against ARID1A (1:300, CUSABIO TECHNOLOGY), βactin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd).

Techniques: Quantitative Proteomics, Expressing, MANN-WHITNEY, RNA Sequencing, Western Blot

Journal: Frontiers in Oncology

Article Title: Downregulated ARID1A by miR-185 Is Associated With Poor Prognosis and Adverse Outcomes in Colon Adenocarcinoma

doi: 10.3389/fonc.2021.679334

Figure Lengend Snippet: COAD patients with low ARID1A mRNA levels had a poorer prognosis and predictive value in the pathological stages of COAD. (A) Prognostic value of ARID1A in primary colon adenocarcinoma using Kaplan-Meier plotter from GEPIA2, OS, Overall survival, DFS, Disease free survival; (B) Prognostic value of ARID1A in metastases using Kaplan-Meier plotter from CanEvolve: M-OS, Metastases overall survival, M-DSS, disease-specific survival, and M-DFS, Metastases disease-free survival; (C) Pathologic M, pathologic N, and pathologic T, respectively; (D) Lymphatic invasion, microsatellite instability, and Vital status, respectively. * p < 0.05 and *** p < 0.001.

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline (1× TBS) containing 0.05% Tween-20, the membrane was incubated overnight at 4°C with rabbit antibodies against ARID1A (1:300, CUSABIO TECHNOLOGY), βactin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd).

Techniques:

Journal: Frontiers in Oncology

Article Title: Downregulated ARID1A by miR-185 Is Associated With Poor Prognosis and Adverse Outcomes in Colon Adenocarcinoma

doi: 10.3389/fonc.2021.679334

Figure Lengend Snippet: Mir-185 downregulates ARID1A in colon cancer. (A) ARID1A can be controlled by mir-185-5p; (B) Transcription factors that regulate ARID1A expression from Cistrome (Chip-Seq data); (C) Mir-185 is overexpressed in COAD; (D) ARID1A is negatively correlated with mir-185-5p in COAD; (E, F) Mir185 mimics and inhibitors results detected by qPCR; (G) Mir185 mimics and inhibitors results detected by western blot.

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline (1× TBS) containing 0.05% Tween-20, the membrane was incubated overnight at 4°C with rabbit antibodies against ARID1A (1:300, CUSABIO TECHNOLOGY), βactin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd).

Techniques: Expressing, ChIP-sequencing, Western Blot

Journal: Frontiers in Oncology

Article Title: Downregulated ARID1A by miR-185 Is Associated With Poor Prognosis and Adverse Outcomes in Colon Adenocarcinoma

doi: 10.3389/fonc.2021.679334

Figure Lengend Snippet: The correlation between ARID1A and immune cell infiltration. (A) The significant positive correlation between ARID1A and subtypes of immune cells (TIMER); (B) Kaplan-Meier curves for the immune infiltrates and ARID1A Expression. The correlation between ARID1A expression and the abundance of CD4+ T cells, B cells, and natural killer cells, respectively (TIMER).

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline (1× TBS) containing 0.05% Tween-20, the membrane was incubated overnight at 4°C with rabbit antibodies against ARID1A (1:300, CUSABIO TECHNOLOGY), βactin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Downregulated ARID1A by miR-185 Is Associated With Poor Prognosis and Adverse Outcomes in Colon Adenocarcinoma

doi: 10.3389/fonc.2021.679334

Figure Lengend Snippet: Correlation analysis between ARID1A and gene biomarkers of immune cells in COAD (TIMER).

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline (1× TBS) containing 0.05% Tween-20, the membrane was incubated overnight at 4°C with rabbit antibodies against ARID1A (1:300, CUSABIO TECHNOLOGY), βactin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: Primer lists and their sequences used in the study

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Sequencing, Negative Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: Identified differentially expressed genes (DEGs)in HCT116‐ARID1A‐wild‐type vs. HCT116‐ARID1A‐knockout (A) Volcano plot of differentially expressed genes (B) Heat map of DEGs in control and knockout groups (C) upper part,KEGG and GO enrichment analysis of differentially upregulated genes. (C) lower part ,KEGG and GO enrichment analysis of differentially downregulated genes

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Knock-Out, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: Expression of EMT‐related markers (VIM) and correlation with ARID1A (A) Vimentin (VIM) overexpression was found in KO cell lines (B) VIM high expression is associated with a poor prognosis in COAD (C) VIM displays high expression in cells with lower ARID1A expression (SW620&RKO) and low expression in cells with a high ARID1A expression (HCT116&LoVo) (D) colon cancer samples were examined for co‐expression of Vimentin and ARID1A (E) GEPIA2 database visualization of the co‐expression of Vimentin and ARID1A in colon cancer tissue.

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Expressing, Over Expression

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: Expression of EMT‐related marker (CDH1) and correlation with ARID1A (A) CDH1 is downregulated in ARID1A deficient cells (B) shows a Kaplan–Maier result of CDH1 in COAD (C and D) E‐cadherin expression correlates positively with ARID1A expression in COAD

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Expressing, Marker

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: ARID1A sequence and mutation profiles in CRC cells used in the research

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Sequencing, Mutagenesis, Variant Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: ARID1A knockdown in HCT116 cell line (A) At 50 nM, 4 μl siRNA‐ARID1A, qPCR demonstrates no change of vimentin and E‐cadherin expression levels in HCT116 after 24 h transfection (B) ARID1A deficiency at 75 nM, 6 μl siRNA‐ARID1A for 24 h activates VIM and suppress CDH1 in HCT116 (C) qPCR results of ARID1A downregulation using 75 nM, 6‐μl siRNA for 48 h showed VIM and CDH1 expression in HCT116. ARID1A silencing altered VIM and E‐cadherin expression (D and E) VIM and CDH1 expression were altered in qPCR results of LS174‐T‐ARID1A knocked down for 24 and 48 h, respectively. ns, Non‐significant; *p < 0.05, **p < 0.001, and ***p < 0.0001

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Knockdown, Expressing, Transfection

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: ARID1A silencing alters VIM and E‐cadherin expression (A and B) VIM and CDH1 expression were altered in WB results after ARID1A knocked down for 24 and 48 h.

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: ARID1A knockdown promotes colon cell line migration and proliferation (A and B) The effect of ARID1A depletion on HCT116 and LS174T cell migration (C) The effect of ARID1A depletion on LS174T cell invasion (D) The proliferation of HCT116 cells was assessed at specified times following ARID1A siRNA transfection (E) Immunoprecipitated DNA from ChIP experiments with anti‐ARID1A antibody was examined by real‐time qPCR using E‐cadherin and Vimentin‐specific promoter primers. IP stands for Immunoprecipitated. *p < 0.05, **p < 0.001, and ***p < 0.0001. Statistical differences were calculated using an imageJ software.

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: Knockdown, Migration, Transfection, Immunoprecipitation, Software

Journal: Journal of Cellular and Molecular Medicine

Article Title: ARID1A downregulation promotes cell proliferation and migration of colon cancer via VIM activation and CDH1 suppression

doi: 10.1111/jcmm.17590

Figure Lengend Snippet: List of 87 ARID1A interaction proteins in HCT116

Article Snippet: The membrane was incubated overnight at 4°C with rabbit antibodies against β‐actin (1:1000, Beijing Zhongshan Biotechnology Co., Ltd); ARID1A (1:300, CUSABIO TECHNOLOGY LLC); E‐cadherin Polyclonal antibody, cat on: 20874‐1‐AP; and Vimentin Polyclonal antibody, cat no: 10366‐1‐AP after blocking with 5% non‐fat milk in Tris‐buffered saline containing 0.05%Tween‐20 (1–1000).

Techniques: