Journal: Cancers

Article Title: The m 7 G Reader NCBP2 Promotes Pancreatic Cancer Progression by Upregulating MAPK/ERK Signaling

doi: 10.3390/cancers15225454

Figure Lengend Snippet: NCBP2 is significantly expressed in PDAC patients and correlated with poor prognosis. ( A ) Analysis of NCBP2 transcript levels in PDAC and normal tissues based on the RNA-seq data from GEPIA 2.0 cohort. ( B ) Analysis of NCBP2 transcript levels in PDAC and normal tissues based on DNA microarray data from GEO cohorts (GSE15471, GSE28735, GSE16515). ( C , D ) Overall survival and disease-free survival curves for PDAC patients with high or low NCBP2 expression in GEPIA 2.0 cohort. ( E , F ) The RNA and protein expression level of NCBP2 in HPDE and five PDAC cell lines. ( G ) The expression level of NCBP2 in PDAC and para-carcinoma tissues from tissue microarray, * p < 0.05 and *** p < 0.001.

Article Snippet: We obtained human embryonic kidney epithelial cell line 293T (HEK293T) and PDAC cell lines (Panc 05.04, PANC-1, AsPC-1, BxPC-3, MIA PaCa-2) from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: RNA Sequencing, Microarray, Expressing

Journal: Cancers

Article Title: The m 7 G Reader NCBP2 Promotes Pancreatic Cancer Progression by Upregulating MAPK/ERK Signaling

doi: 10.3390/cancers15225454

Figure Lengend Snippet: NCBP2 promotes PDAC cell growth in vitro. ( A – C ). The cell counting, 3D sphere, and colony formation assay results in control and NCBP2-knockdown Panc 05.04 and PANC-1 cells. ( D – F ). The cell counting, 3D sphere and colony formation assay results in control and NCBP2-overexpression AsPC-1 and BxPC-3 cells, * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: We obtained human embryonic kidney epithelial cell line 293T (HEK293T) and PDAC cell lines (Panc 05.04, PANC-1, AsPC-1, BxPC-3, MIA PaCa-2) from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vitro, Cell Counting, Colony Assay, Control, Knockdown, Over Expression

Journal: Cancers

Article Title: The m 7 G Reader NCBP2 Promotes Pancreatic Cancer Progression by Upregulating MAPK/ERK Signaling

doi: 10.3390/cancers15225454

Figure Lengend Snippet: NCBP2 promotes PDAC cell growth in-vivo. ( A , B ) Tumor growth curves of xenograft models established from control or stable NCBP2-knockdown Panc 05.04 cells. ( C ) Assessment of tumor weight from control and NCBP2-knockdown groups. ( D , E ) The expression level of Ki-67 in tumor tissues from control and NCBP2-knockdown groups. ( F ) Body weight of nude mice in control and NCBP2-knockdown group, * p < 0.05, and ** p < 0.01.

Article Snippet: We obtained human embryonic kidney epithelial cell line 293T (HEK293T) and PDAC cell lines (Panc 05.04, PANC-1, AsPC-1, BxPC-3, MIA PaCa-2) from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vivo, Control, Knockdown, Expressing

Journal: Cancers

Article Title: The m 7 G Reader NCBP2 Promotes Pancreatic Cancer Progression by Upregulating MAPK/ERK Signaling

doi: 10.3390/cancers15225454

Figure Lengend Snippet: NCBP2 upregulates c-JUN to activate MEK/ERK signaling in a m 7 G-dependent manner. ( A ) KEGG analysis of RNA-seq data in PDAC patients with high or low NCBP2 expression from TCGA cohort. ( B ) The GSEA enrichment plot of “MAPK signaling pathway” in PDAC patients with high or low NCBP2 expression from TCGA cohort. ( C ) Immunoblotting for protein levels of total JNK/phosphorylated JNK (Thr183/Tyr185), total p38/phosphorylated p38 (Thr180/Tyr182), and total ERK/phosphorylated ERK (Thr202/Tyr204) in control and NCBP2-knockdown PDAC cells. Tubulin was used as the internal control. ( D ) Immunoblotting for protein levels of total/phosphorylated MEK and total/phosphorylated ERK (Thr202/Tyr204) in control and c-JUN-knockdown Panc 05.04 and PANC-1 cells. Tubulin was used as the internal control. ( E ) The mRNA expression levels of NCBP2 and c-JUN in control and NCBP2-knockdown Panc 05.04 and PANC-1 cells. ( F ) Immunoblotting for protein levels of NCBP2 and c-JUN in control and NCBP2-knockdown Panc 05.04 and PANC-1 cells. Tubulin was used as the internal control. ( G ) Polysome profiling results of the control and NCBP2-knockdown Panc 05.04 and PANC-1 cells. ( H ) Gene-specific m 7 G qPCR results for the m 7 G methylation levels of c-JUN in PANC 05.04 and PANC-1 cells, *** p < 0.001.

Article Snippet: We obtained human embryonic kidney epithelial cell line 293T (HEK293T) and PDAC cell lines (Panc 05.04, PANC-1, AsPC-1, BxPC-3, MIA PaCa-2) from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: RNA Sequencing, Expressing, Western Blot, Control, Knockdown, Methylation

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A Experimental procedure of the GEM-treated PDX PDAC model. B Immunoblotting assay to detect the expression of cyclin and cyclin-dependent kinases (Cyclin D1, Cyclin E, CDK2, CDK4) at the G1/S cell cycle checkpoint and cancer stem cell master genes (CD133, OCT4, NANOG, and Sox2) in tissues from the 3-generation PDX mice that treated with saline or gemcitabine. C Heatmap shows differentially expressed lncRNAs in GEM-treated and control PDX mice. D Comparison of expression of lncRNAs in tumor tissues from 36 PDAC patients with paired paracancerous tissues. E Correlation analysis of LINC01134 with cyclin and cyclin-dependent kinases and cancer stem cell master genes as described in ( B ). F Comparison of the expression of LINC01134 in tumor tissues and paired paraneoplastic tissues from 70 PDAC patients. G Kaplan–Meier analysis of the correlation between expression levels of LINC01134 and PDAC prognosis in 57 patients. Data are expressed as mean ± SD * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Western Blot, Expressing, Saline, Control, Comparison

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A Statistical analysis of the primary and secondary sphere formation ability of PDAC cells, and representative images show secondary sphere formation in these cells. B Western blot was performed to detect the effects of silencing or overexpression of LINC01134 on the expression of the stemness-related genes. C qRT-PCR detects the effect of silencing or overexpression of LINC01134 on the expression of the stemness-related genes. D Effects of silencing LINC01134 on transplanted tumors in nude mice. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Western Blot, Over Expression, Expressing, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A Colony formation assay of cells containing silenced or overexpressed LINC01134 after GEM treatment. B Effects of different concentrations of GEM on the viability of PDAC cells. C Expression of Ki67 in xenograft tumors formed from PDAC cells transfected with Ctrl or shLINC01134 in GEM-treated nude mice. D Typical images of xenograft tumors formed from PDAC cells transfected with Ctrl or shLINC01134 in GEM-treated nude mice. Tumor growth curves of PDAC cells transfected with Ctrl or shLINC01134 in GEM-treated nude mice. E Expression of cyclin and cyclin-dependent kinases and and cancer stem cell master genes in xenograft tumor tissues formed from PDAC cells that transfected with Ctrl or shLINC01134 in GEM-treated nude mice. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Colony Assay, Expressing, Transfection

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A FISH assay detects the localization of LINC01134 in PDAC cells. B qRT-PCR detects the distribution of LINC01134 in nuclear and cytoplasm in PDAC cells. C Wayne diagram shows microRNAs predicted by the bioinformatics databases Encyclopedia of RNA Interactomes and lncRNASNP2 to target bind to LINC01134. D RNA pull-down analysis shows that miR-140-3p was highly enriched in LINC01134 precipitates. E Correlation analysis shows a significant negative correlation between LINC01134 and miR-140-3p in PDAC. F Dual-luciferase reporter assay detects the relative activity of luciferase in transfected PDAC cells. G RIP assay detects the relative enrichment of LINC01134 and miR-140-3p in anti-IgG or anti-AGO2 specific immunoprecipitates. H RNA pull-down assay was used to detect the inter-relationship between LINC01134, miR-140-3p, and AGO2. I Effects of silencing or overexpression of LINC01134 on the expression of miR-140-3p. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Quantitative RT-PCR, Luciferase, Reporter Assay, Activity Assay, Transfection, Pull Down Assay, Over Expression, Expressing

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A Heatmap and volcano map showing differentially expressed genes in PDAC after the overexpression of miR-140-3p. B KEGG pathway analysis of the major pathways associated with differentially expressed genes. C Correlation analysis shows that miR-140-3p was negatively correlated with WNT5A. D Dual-luciferase reporter analysis detects the relative activity of luciferase in post-transfected PDAC cells. E , F Effects of silencing or overexpression of miR-140-3p on the expression of WNT5A in PDAC cells. G Effects of co-transfection of LINC01134 and miR-140-3p on the expression of WNT5A in PDAC cells. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Over Expression, Luciferase, Activity Assay, Transfection, Expressing, Cotransfection

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A Correlation analysis shows that the abundance of LINC01134 was positively correlated with WNT5A. B CCK-8 analysis of the effects of silencing or overexpressing LINC01134 and overexpressing or silencing WNT5A when co-transfected on the viability of PDAC cells. C Representative phase contrast images of tumorspheres formed by PDAC cells transduced with indicated constructs or a nontarget shRNA. D Statistical analysis of the primary and secondary sphere formation ability of PDAC cells as described in ( C ). E Effects of silencing or overexpression of LINC01134 and overexpression or silencing of WNT5A on the expression of the stemness-related genes when co-transfected. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: CCK-8 Assay, Transfection, Transduction, Construct, shRNA, Over Expression, Expressing

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A Silencing LINC01134 reduced the level of WNT5A and its downstream signaling activity in PDAC cells, and overexpression of LINC01134 enhanced the level of WNT5A and its downstream signaling activity in PDAC cells. B Overexpression of LINC01134 promoted the growth of transplanted tumors in PDAC nude mice and induced resistance to GEM. At the same time, the application of salinomycin effectively antagonized the pro-tumorigenic effect of LINC01134 and restored the sensitivity of transplanted tumors to GEM treatment. C IHC assays indicate the expression of Cyclin D1, Cyclin E, CDK2, CD133, Ki67, OCT4, NANOG and Sox2. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Activity Assay, Over Expression, Expressing

Journal: Cell Death & Disease

Article Title: FTO-mediated LINC01134 stabilization to promote chemoresistance through miR-140-3p/WNT5A/WNT pathway in PDAC

doi: 10.1038/s41419-023-06244-7

Figure Lengend Snippet: A The results of MeRIP-qPCR indicate an overall lower level of m6A in BxPC-3 and PANC-1 cells than in HPDE6-C7 cells. B Analysis of the expression of FTO in PDAC using the GEPIA database. C Correlation analysis shows that the expression of FTO was significantly and positively correlated with the abundance of LINC01134. D Effects of silencing FTO on the expression of LINC01134. E Effects of overexpression of FTO on the expression of LINC01134. F An online engine ( http://m6avar.renlab.org/ ) can be used to predict the m6A modification site of LINC01134 and the prediction was confirmed by a luciferase analysis report. G Effects of silencing or overexpression of FTO on the degree of m6A enrichment of LINC01134. H Effects of silencing or overexpression of FTO on the stability of LINC01134 in the presence of radio-luciferin D. I RIP assay confirms the binding between YTHDF2 and LINC01134. J Effects of silencing YTHDF2 on the expression of LINC01134. K Effects of silencing YTHDF2 on the stability of LINC01134 in the presence of radio-labeled D. L RIP experiments confirm the effects of silencing FTO on the expression of YTHDF2. Data are expressed as mean, * p < 0.05, ** p < 0.01.

Article Snippet: The standard human pancreatic cell line HPDE6-C7 and human PDAC cell line CaPan-1 were purchased from the American Tissue Culture Colection (ATCC), and the other human PDAC cell lines PANC-1, MIA PaCa-2, BxPC-3, SW1990, CFPAC-1, and AsPC-1 were purchased from the Cell Bank of the Typical Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Expressing, Over Expression, Modification, Luciferase, Binding Assay, Labeling

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: Low 15‐hydroxyprostaglandin dehydrogenase (15‐ PGDH ) expression is implicated in poor pancreatic ductal adenocarcinoma ( PDAC ) prognosis. A, Workflow diagram of patients who underwent pancreatic resection and contributed samples for immunohistochemical ( IHC ) analysis. B, Representative IHC staining of 15‐ PGDH expression in 107 PDAC tissues. Scale bar = 100 μm. C,D, Relationship between 15‐ PGDH expression and relapse‐free survival (C) or overall survival (D) using the Kaplan‐Meier method

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: 15‐Hydroxyprostaglandin dehydrogenase (15‐ PGDH ) downregulation by interleukin‐1β ( IL ‐1β) enhances pancreatic ductal adenocarcinoma cell growth. A,B, Expression of HPGD (the gene coding 15‐ PGDH protein, upper panel) or 15‐ PGDH (lower panel) in PK ‐8 cells (A) or S2‐013 cells (B) after treatment with si RNA targeting 15‐ PGDH or with control si RNA , evaluated by quantitative RT ‐ PCR (upper panel) or Western blot analysis (lower panel). Data are presented as the treated/control cell ratio. C,D, PK ‐8 cells (C) or S2‐013 cells (D) transfected with si RNA s targeting 15‐ PGDH or with control si RNA were incubated for up to 96 hours and assayed for cell number; data are presented as the treated/control (time = 0) cell ratio. E,F, Expression of 15‐ PGDH in PK ‐8 cells or S2‐013 cells after IL ‐1β (E) or tumor necrosis factor‐α ( TNF ‐α) (F) treatment for 24 and 48 hours and distilled water treatment for 48 hours as a control was evaluated by Western blotting. G, Column graph showing relative 15‐ PGDH levels in PK ‐8 cells or S2‐013 cells after IL ‐1β and TNF ‐α treatment for 24 and 48 hours, and distilled water treatment for 48 hours as a control, were evaluated using ImageJ software. H, Expression of HPGD and IL 1B in six PDAC patients determined by quantitative RT ‐ PCR . Data were normalized to the ACTB mRNA level and are shown as the mean ± SD of three independent experiments. **P < .01

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Transfection, Incubation, Software

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: Tumor‐associated macrophages are inversely correlated with pancreatic ductal adenocarcinoma ( PDAC ) cells harboring high 15‐hydroxyprostaglandin dehydrogenase (15‐ PGDH ) expression. A,B, Representative immunohistochemical ( IHC ) staining of 15‐ PGDH (upper panel) and CD 163 (lower panel) expression in high 15‐ PGDH (A) and low 15‐ PGDH (B) serial PDAC specimens. Scale bar = 200 μm. C, Graph showing Pearson's correlation between the expression of 15‐ PGDH and the number of CD 163‐positive cells in 107 PDAC patients. D, Schematic representation of the findings of this study. IL ‐1βR, interleukin‐1β receptor

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry