Journal: Oncotarget

Article Title: KHSRP-bound small nucleolar RNAs associate with promotion of cell invasiveness and metastasis of pancreatic cancer.

doi: 10.18632/oncotarget.27413

Figure Lengend Snippet: Figure 1: KHSRP distribution in PDAC cells. S2-013 (A) and PANC-1 (B) cells were incubated on fibronectin and immunocytochemically labeled with anti-KHSRP antibody (green). Actin filaments were labeled by phalloidin (red). Arrows, KHSRP localized in cell protrusions. Blue, DAPI staining. Bars, 10 µm.

Article Snippet: The human PDAC cell line PANC-1 was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Incubation, Labeling, Staining

Journal: Oncotarget

Article Title: KHSRP-bound small nucleolar RNAs associate with promotion of cell invasiveness and metastasis of pancreatic cancer.

doi: 10.18632/oncotarget.27413

Figure Lengend Snippet: Figure 2: KHSRP promotes cell motility and invasion of PDAC cells. (A) Effect of KHSRP siRNA in S2-013 cells. Western blots probed with anti-KHSRP antibody show two S2-013 KHSRP RNAi clones (siKH-1 and -2) transfected with siRNA targeting KHSRP and two scrambled control RNAi clones (Scr-1 and -2). (B) Control RNAi or KHSRP RNAi S2-013 cells were seeded into two-chamber motility chambers. Migrating cells in four fields per group were counted. Data are derived from three independent experiments and expressed as mean ± SD. *p < 0.05 compared with Scr-1 and Scr-2 (Student’s t-test). (C) Control RNAi or KHSRP RNAi S2-013 cells were seeded into Matrigel invasion chambers. Invading cells in four fields per group were counted. Data are derived from three independent experiments and expressed as mean ± SD. *p < 0.05 compared to Scr-1 or Scr-2 (Student’s t-test). (D) The mock control vector or myc-tagged KHSRP- rescue construct was transiently transfected into control RNAi and KHSRP RNAi cells. Western blotting was performed using anti-KHSRP and anti-myc antibodies. Closed arrowhead, endogenous KHSRP; open arrowhead, exogenous KHSRP. Closed arrow head, endogenous KHSRP; open arrow head, exogenous KHSRP. (E, F) The mock control vector or myc-tagged KHSRP-rescue construct was transiently transfected into control RNAi and KHSRP RNAi cells; 48 h later, motility (E) and two-chamber invasion (F) assays were performed. Migrating cells in four fields per group were counted. Data are derived from three independent experiments and expressed as mean ± SD. *p < 0.05 compared with corresponding siKH-1 and siKH-2 transfected mock vector (Student’s t-test).

Article Snippet: The human PDAC cell line PANC-1 was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Western Blot, Clone Assay, Transfection, Control, Derivative Assay, Plasmid Preparation, Construct

Journal: Oncotarget

Article Title: KHSRP-bound small nucleolar RNAs associate with promotion of cell invasiveness and metastasis of pancreatic cancer.

doi: 10.18632/oncotarget.27413

Figure Lengend Snippet: Figure 9: Knockdown effect of siRNA-FA-PEG-COL nanoparticles targeting KHSRP-bound snoRNAs on cell motility and invasion in the orthotopic murine model of PDAC. (A) Development of carcinomatosis in S2-013 tumor-bearing mice treated with scrambled control siRNA-FA-PEG-COL nanoparticles (Scr) and target siRNA-FA-PEG-COL nanoparticles against SNORA18 (si-snora18) and SNORA22 (si-snora22). Arrow, primary tumor; arrowheads, dissemination nodules in the abdominal cavity. (B–D) Hematoxylin and eosin staining of representative sections of S2-013-derived PDAC tumor tissues in mice treated with scrambled control siRNA-FA-PEG-COL nanoparticles showing areas of regional invasion of the retroperitoneum (B) and distant metastases to the lung (C) and liver (D). Original magnification: 200 ×.

Article Snippet: The human PDAC cell line PANC-1 was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Knockdown, Control, Staining, Derivative Assay

Journal: Translational Oncology

Article Title: The pancreatitis-cancer transformation-related factor, human rhomboid family-1, promotes pancreatic cancer progression through the SRC/YAP signaling pathway

doi: 10.1016/j.tranon.2025.102346

Figure Lengend Snippet: Increased RHBDF1 levels in chronic pancreatitis linked to PDAC are associated with accelerated disease progression. (A) Transcriptomic analysis of tumor and surrounding tissues from FUSCC revealed significant differences in the level of RHBDF1 mRNA expression between CP-PDAC and adjacent normal tissues ( N = 11, P < 0.001). (B) Transcriptomic analysis of TGCA and GTEx databases revealed significant differences in the level of RHBDF1 mRNA expression between PDAC and normal pancreas tissues. (C) Kaplan–Meier plots illustrating the relationship between RHBDF1 expression and overall survival from TCGA data. (D) Western blot analysis of RHBDF1 in pancreatic ductal adenocarcinoma cell lines and CP, CP-PDAC, and adjacent tissues. (E, F) IHC staining analysis show the relative levels of RHBDF1 in CP, CP-PDAC, and adjacent tissue. (G, H) Immunohistochemical analysis of RHBDF1 expression in LSL-KrasG12D and Ptf1aCreER mice with chronic pancreatitis induced by cerulein. *P <0.05, **P <0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Human PDAC cells (Capan-1, MiaPaCa-2, SW1990, PANC-1, and BxPC-3), human pancreatic ductal epithelial cells (HPDE), human pancreatic stellate cells (HPSCs), and cancer-associated fibroblasts (CAFs) were obtained from the American Type Culture Collection.

Techniques: Biomarker Discovery, Expressing, Western Blot, Immunohistochemistry, Immunohistochemical staining

Journal: Translational Oncology

Article Title: The pancreatitis-cancer transformation-related factor, human rhomboid family-1, promotes pancreatic cancer progression through the SRC/YAP signaling pathway

doi: 10.1016/j.tranon.2025.102346

Figure Lengend Snippet: RHBDF1 increases the proliferation, migration, and invasion of pancreatic tumor cells. (A-C) Three short hairpin RNAs (shRNAs) aimed at the RHBDF1 junction site were designed. Following stable transfection with sh-RHBDF1, the levels of RHBDF1 mRNA and protein in PDAC cells were measured by qRT-PCR and western blotting. (D–F) Cell proliferation was assessed by the CCK-8 and EdU incorporation assays. Scale bars, 100 μm. (G-I) The cell cycle was evaluated using flow cytometry. (J, K) Transwell assays were performed to assess the migration and invasion abilities of PDAC cells. Scale bars, 100 μm. * P <0.05, ** P <0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human PDAC cells (Capan-1, MiaPaCa-2, SW1990, PANC-1, and BxPC-3), human pancreatic ductal epithelial cells (HPDE), human pancreatic stellate cells (HPSCs), and cancer-associated fibroblasts (CAFs) were obtained from the American Type Culture Collection.

Techniques: Migration, Stable Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: Translational Oncology

Article Title: The pancreatitis-cancer transformation-related factor, human rhomboid family-1, promotes pancreatic cancer progression through the SRC/YAP signaling pathway

doi: 10.1016/j.tranon.2025.102346

Figure Lengend Snippet: RHBDF1 activates SRC to exert carcinogenic effects. (A) Volcano plot illustrating the differentially expressed genes (DEGs) between the RHBDF1-high and RHBDF1-low groups analyzed using DESeq2. |Log2fold change| > 1, FDR < 0.05. (B) Protein-protein internation network of overexpressed mRNAs in the RHBDF1-high group, as constructed by Network Analyst. The size of each mRNA represents their topology level in the network. (C–D) RHBDF1 and SRC are co-expressed at the mRNA level in the CCLE (C) and TCGA datasets (D). (E–M) The expression levels of RHBDF1 and SRC mRNA and protein in PDAC cells were measured by qRT-PCR and western blotting. (N) Co-immunoprecipitation of RHBDF1 and SRC in PDAC cells. (O–Q) Cell growth was evaluated using the CCK-8 and EdU incorporation tests. Scale bars, 100 μm. (R, S) The migration and invasion potential of PDAC cells was assessed using the Transwell assay. Scale bars, 100 μm. * P <0.05, ** P <0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human PDAC cells (Capan-1, MiaPaCa-2, SW1990, PANC-1, and BxPC-3), human pancreatic ductal epithelial cells (HPDE), human pancreatic stellate cells (HPSCs), and cancer-associated fibroblasts (CAFs) were obtained from the American Type Culture Collection.

Techniques: Construct, Expressing, Quantitative RT-PCR, Western Blot, Immunoprecipitation, CCK-8 Assay, Migration, Transwell Assay

Journal: Translational Oncology

Article Title: The pancreatitis-cancer transformation-related factor, human rhomboid family-1, promotes pancreatic cancer progression through the SRC/YAP signaling pathway

doi: 10.1016/j.tranon.2025.102346

Figure Lengend Snippet: RNHBDF1 activates the YAP signaling pathway through SRC. (A) Differences between the shRHBDF1 and shNC groups were analyzed by principal component analysis and volcano plot analysis. (B) The differentially expressed genes (DEGs) in the two sets of transcriptomics data (clinical samples and cell line RHBDF-1 knockdown) were performed. (C, D) Gene Ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. (E-H) Western blot analysis was employed to confirm the expression levels of SRC/YAP key proteins in ShNC, shRHBDF1, and shRHBDF1 combined with SRC inhibition groups. (I–K) Cell proliferation was assessed using the CCK-8 and EdU incorporation assays. Scale bars, 100 μm. (L, M) The migration and invasion abilities of PDAC cells were assessed using the Transwell assay. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human PDAC cells (Capan-1, MiaPaCa-2, SW1990, PANC-1, and BxPC-3), human pancreatic ductal epithelial cells (HPDE), human pancreatic stellate cells (HPSCs), and cancer-associated fibroblasts (CAFs) were obtained from the American Type Culture Collection.

Techniques: Knockdown, Functional Assay, Western Blot, Expressing, Inhibition, CCK-8 Assay, Migration, Transwell Assay

Journal: International journal of cancer

Article Title: Nuclear expression of Y-box binding protein-1 is associated with poor prognosis in patients with pancreatic cancer and its knockdown inhibits tumor growth and metastasis in mice tumor models.

doi: 10.1002/ijc.30075

Figure Lengend Snippet: Figure 2. Effects of YB-1 knockdown on proliferation and cell cycle state in PDAC cells. The levels of YBX1 mRNA (a) and YB-1 protein (b) expression were assessed by quantitative RT-PCR and western blot analyses in PDAC and non-cancerous HPDE cells. Fluorescence immuno- cytochemical staining (c) reveals nuclear YB-1 expression in all PDAC cell lines, but not in the HPDE cell line. (d–g) PDAC cells were treated with YB-1 or control siRNAs (20 nM) for 48 h, and subjected to quantitative RT-PCR and western blot analyses for YB-1 expression, WST-8 assay for cell proliferation and flow cytometry for cell cycle analysis. The expressions of YBX1 mRNA (d) and YB-1 protein (e), and cell prolif- eration (f) were significantly lower in the YB-1 siRNAtransfected groups compared with the control group in all PDAC cell lines. The propor- tion in G0/G1 phase (g) was increased in all PDAC cells with YB-1 siRNAs treatment. Data represent means 6 SD (n 5 3 for each). *p < 0.05.

Article Snippet: Human PDAC cell lines (AsPC-1, MIA PaCa-2, PANC-1) were purchased from the American Type Culture Collection.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Fluorescence, Staining, Control, Cytometry, Cell Cycle Assay

Journal: International journal of cancer

Article Title: Nuclear expression of Y-box binding protein-1 is associated with poor prognosis in patients with pancreatic cancer and its knockdown inhibits tumor growth and metastasis in mice tumor models.

doi: 10.1002/ijc.30075

Figure Lengend Snippet: Figure 3. Effects of YB-1 knockdown on cell cycle arrest and responsible factors in PDAC cells. PDAC cells were treated withYB-1 or control siR- NAs (20 nM) for 48 h, and subjected to western blot and quantitative RT-PCR analyses for expression of cell cycle-related factors, and flow cytometer for cell cycle analysis. As shown in representative immunoblots (a: left panel), YB-1 knockdown induced the decreases in CDK2 and SKP2 proteins and the increase in p27 protein (a: right panel), and decreased the SKP2 mRNA (b) in all PDAC cell lines. As shown in the rep- resentative immunoblots (c: left panel), the transfection of the CDK2, SKP2, and p27 siRNAs significantly downregulated each target protein expression, and SKP2 knockdown also induced a significant increase in p27 protein expression (c: right panel). (d) CDK2 knockdown did not induce inhibition of the G1/S phase transition. In contrast, SKP2 knockdown increased the cell population in G0/G1 phase with accumulation of p27 protein, while double-knockdown of SKP2 and p27 decreased the cell population in G0/G1 phase concomitant with the increased pop- ulation in S phase compared with single SKP2 knockdown. Data represent means 6 SD (n 5 3 for each). *p < 0.05.

Article Snippet: Human PDAC cell lines (AsPC-1, MIA PaCa-2, PANC-1) were purchased from the American Type Culture Collection.

Techniques: Knockdown, Control, Western Blot, Quantitative RT-PCR, Expressing, Cytometry, Cell Cycle Assay, Transfection, Inhibition, Sublimation

Journal: International journal of cancer

Article Title: Nuclear expression of Y-box binding protein-1 is associated with poor prognosis in patients with pancreatic cancer and its knockdown inhibits tumor growth and metastasis in mice tumor models.

doi: 10.1002/ijc.30075

Figure Lengend Snippet: Figure 4. Effects of YB-1 knockdown on invasive ability and MMP expressions in PDAC cells. PDAC cells were transfected with the indicated siRNAs (20 nM) for 48 h, and subjected to Matrigel invasion assay, and quantitative RT-PCR and western blot analyses for MMP expres- sions. As shown in the representative images (a: left panel), the invasive ability was significantly suppressed in YB-1 siRNA-transfected cells compared with the controls (a: right panel). Western blot and quantitative RT-PCR analyses show the decreases in MT2-MMP protein (b, c) and mRNA (d) expressions after transfection of the YB-1 siRNAs. The invasive ability (e) was significantly suppressed by MT2-MMP siRNA transfection compared with the control in all PDAC cell lines. Data represent means 6 SD (n 5 3 for each). *p < 0.05. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: Human PDAC cell lines (AsPC-1, MIA PaCa-2, PANC-1) were purchased from the American Type Culture Collection.

Techniques: Knockdown, Transfection, Invasion Assay, Quantitative RT-PCR, Western Blot, Control

Journal: International journal of cancer

Article Title: Nuclear expression of Y-box binding protein-1 is associated with poor prognosis in patients with pancreatic cancer and its knockdown inhibits tumor growth and metastasis in mice tumor models.

doi: 10.1002/ijc.30075

Figure Lengend Snippet: Figure 5. Effects of YB-1 knockdown on tumor growth and liver metastasis in vivo. Suit-2-luc and MIA PaCa-2-luc cells were transfected with control siRNA or YB-1 siRNA#1 (20 nM) ex vivo and then transplanted into the subcapsular region of the spleen in nude mice. (a) Representa- tive IVIS images for tumor growth of Suit-2-luc and MIA PaCa-2-luc cells at the indicated time points in the control or YB-1 siRNA#1 groups. The total tumor volumes evaluated by luciferase activity were significantly inhibited on Day 22 in the YB-1 siRNA#1 groups (n 5 17 for Suit-2- luc; n 5 7 for MIA PaCa-2-luc) compared with the control groups (n 5 18 and n 5 7, respectively). Data represent means6 SD. (b) Macroscopic observation by laparotomy on Day 29 shows massive tumor formation in the liver (arrow) and spleen (arrowhead) in the control group, but not obviously in the YB-1 siRNA#1 group (left panel). The area surrounded by the dotted line (a) is defined as the metastatic hepatic area. The volumes of liver metastasis were significantly inhibited on Day 22 in the YB-1 siRNA#1 group compared with the control group (right panel). (c) Liver metastases were formed in 9 of 18 mice with control siRNA and 3 of 17 mice with the YB-1 siRNA#1 for Suit-2-luc cells at Day 29 postimplantation, while four of seven in the control siRNA and none of seven mice in the YB-1 siRNA#1 group for MIA PaCa-2-luc cells. There are significant differences in the incidence of liver metastasis between the two groups for both PDAC cells. (d) When subcutaneous tumors of Suit-2-luc were grown to approximate 50 mm3 in volume in nude mice, the YB-1 AON with amido-bridged nucleic acids solutions (200 lg in 200 lL saline) were intravenously administered once a week for 4 weeks. The tumor volumes were significantly inhibited in the YB-1 AON group than the control on 28-day postadministration. Data represent means 6 SD (n 5 5 for each). *p < 0.05. (e) The immunohisto- chemical analysis shows the inhibition of YB-1 expression in tumor tissues with the YB-1 AON treatment compared with the control.

Article Snippet: Human PDAC cell lines (AsPC-1, MIA PaCa-2, PANC-1) were purchased from the American Type Culture Collection.

Techniques: Knockdown, In Vivo, Transfection, Control, Ex Vivo, Luciferase, Activity Assay, Saline, Inhibition, Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Expression of long non-coding RNA LOC285194 and its prognostic significance in human pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: qRT-PCR analysis of lncRNA LOC285194 expression in PDAC cell lines and tissue samples. A. The expression level of LOC285194 in a normal human pancreatic duct epithelial cell line (HPDE6-C7) and PDAC cell lines (PANC-1, BxPC-3, AsPC-1, and PL45). B. The relative level of LOC285194 expression in PDAC tissues and non-tumor tissues. GAPDH was used as an internal control. Results are expressed as mean ± SD for three replicate determination *P < 0.05.

Article Snippet: A normal human pancreatic duct epithelial cell line (HPDE6-C7) and four PDAC cell lines (PANC-1, BxPC-3, AsPC-1, and PL45) were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Quantitative RT-PCR, Expressing, Control

Journal: Cancer Science

Article Title: Tumor microenvironmental 15‐PGDH depletion promotes fibrotic tumor formation and angiogenesis in pancreatic cancer

doi: 10.1111/cas.15495

Figure Lengend Snippet: 15‐PGDH is expressed in the tumor microenvironment and promotes tumorigenesis of PDAC. (A) Representative immunofluorescence staining for 15‐PGDH, AE1/AE3, αSMA and CD45. The arrowheads show strongly 15‐PGDH‐positive cells. Scale bars, 100 μm. (B) Quantification of AE1/AE3 + , αSMA + and CD45 + cells. (C, D) 15‐pgdh expression in 15‐pgdh +/− mice and wild‐type mice was evaluated by qRT‐PCR (C) and western blotting (D). (E) Expression of 15‐pgdh in Panc02 cell lines and normal spleen tissue was evaluated by western blotting. (F) Strategy used to establish the orthotopic transplantation model. Panc02 cells deficient in 15‐pgdh were injected into the pancreas of wild‐type and 15‐pgdh +/− mice to generate orthotopic xenografts. After 21 days, the mice were euthanized, and the tumors were harvested and weighed. (G) Images showing pancreatic tumors and spleens. Scale bars, 1 cm. (H) Main tumor area of wild‐type and 15‐pgdh +/− mice measured by ImageJ software. (I) Volumes of tumors from wild‐type and 15‐pgdh +/− mice. (J) PGE2 expression was measured by LC/MS in wild‐type mice and 15‐pgdh +/− mice. NS, not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The human PDAC cell lines PANC1, PK59, PK‐8, AsPC1, and MIAPaCa2 were obtained from the Japanese Collection of Research Bioresources Cell Bank and RIKEN Bioresource Center Cell Bank.

Techniques: Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Western Blot, Transplantation Assay, Injection, Software, Liquid Chromatography with Mass Spectroscopy

Journal: Cancer Science

Article Title: Tumor microenvironmental 15‐PGDH depletion promotes fibrotic tumor formation and angiogenesis in pancreatic cancer

doi: 10.1111/cas.15495

Figure Lengend Snippet: 15‐pgdh depletion in a highly fibrotic tumor microenvironment maintains angiogenesis levels. (A) Nano‐CT scanning of transplanted tumors from wild‐type and 15‐pgdh +/− mice and quantification of vessel branching. (B) Representative immunohistochemical staining and quantification of CD31 in wild‐type and 15‐pgdh +/− mouse tumors. Scale bars, 500 μm. (C) Expression of VEGFA in wild‐type and 15‐pgdh +/− mouse tumors was determined by qRT‐PCR. (D) Expression of VEGFA in five human PDAC cell lines and three CAF lines was determined by qRT‐PCR. (E) Morphology and quantification of branch points in HUVECs after culture in RPMI 1640 medium and CAF CM. Scale bars, 200 μm. (F) Morphology and quantification of branch points in HUVECs after culture in CAF CM and CAF CM + bevacizumab. Scale bars, 200 μm. (G) PK8 and PK8 cells + CAFs on CAM samples were evaluated for angiogenesis. Scale bars, 1 mm. (H) Immunofluorescence staining for VE‐cadherin and quantification of VE‐cadherin + cells. Scale bars, 200 μm. NS, not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The human PDAC cell lines PANC1, PK59, PK‐8, AsPC1, and MIAPaCa2 were obtained from the Japanese Collection of Research Bioresources Cell Bank and RIKEN Bioresource Center Cell Bank.

Techniques: Immunohistochemical staining, Staining, Expressing, Quantitative RT-PCR, Immunofluorescence

Journal: Cancer Science

Article Title: Tumor microenvironmental 15‐PGDH depletion promotes fibrotic tumor formation and angiogenesis in pancreatic cancer

doi: 10.1111/cas.15495

Figure Lengend Snippet: Genetic deletion of 15‐pgdh favors fibrosis and angiogenesis in a PDAC mouse model. (A) Images showing pancreatic tumors and spleens of KC and PgKC mice. Main pancreas area of KC and PgKC mice measured by ImageJ. Scale bars, 1 cm. (B) H&E staining of KC and PgKC mice, Scale bars, 200 μm. (C) Representative Masson's trichrome staining of KC and PgKC mouse tumors. The graph on the right shows the area of collagen expression. KC: Kras LSL‐G12D/+ ; PgKC, Ptf1aCre /+ mice: 15pgdh −/− ; Kras LSL‐G12D/+ ; Ptf1aCre /+ mice. Scale bars, 500 μm. (D) Representative immunofluorescence staining for αSMA in KC and PgKC mice. The graph on the right shows the quantification of αSMA + cells. Scale bars, 50 μm. (E) Representative immunohistochemical staining for CD31 and quantification of CD31 + cells in KC and PgKC mice, Scale bars, 50 μm. NS, not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The human PDAC cell lines PANC1, PK59, PK‐8, AsPC1, and MIAPaCa2 were obtained from the Japanese Collection of Research Bioresources Cell Bank and RIKEN Bioresource Center Cell Bank.

Techniques: Staining, Expressing, Immunofluorescence, Immunohistochemical staining