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: Cancers

Article Title: A LETM2-Regulated PI3K-Akt Signaling Axis Reveals a Prognostic and Therapeutic Target in Pancreatic Cancer.

doi: 10.3390/cancers14194722

Figure Lengend Snippet: Figure 2. Validation of LETM2 prognostic value and clinical significance for pancreatic cancer in TCGA database. (A) TCGA + GTEx database analysis revealed that LETM2 expression was significantly elevated in PDAC tumor tissue in comparison with normal tissue. Red represents tumor tissue and blue represents normal tissue. (B,C) High levels of LETM2 expression correlated with significantly poor (B) OS and (C) DFS of PDAC patients based on survival analysis of the TCGA database. The dotted lines represented 95% confidence intervals. (D) LETM2 expression correlated with tumor inflammation signature. (E) LETM2 expression correlated with PI3K-AKT- mTOR pathway. (F) LETM2 expression correlated with the P53 pathway. (G) LETM2 expression correlated with immune infiltration level of B cell expression. (H) LETM2 expression correlated with immune infiltration level of T cell CD8+ expression. (I) Sankey diagram demonstrated the association between LETM2 expression and histological grade and living status. * p < 0.05.

Article Snippet: Three kinds of PDAC cell lines (MIA PaCa-2, SW1990, and BxPc-3) and immortalized human pancreatic duct cells (hTERT-HPNE) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Biomarker Discovery, Expressing, Comparison

Journal: Cancers

Article Title: A LETM2-Regulated PI3K-Akt Signaling Axis Reveals a Prognostic and Therapeutic Target in Pancreatic Cancer.

doi: 10.3390/cancers14194722

Figure Lengend Snippet: Figure 4. LETM2 promotes PDAC cell proliferation, migration, and invasion in vitro. (A) Western blotting analysis of LETM2 protein expression in three types PDAC cell lines (MIA PaCa-2, SW1990, and BxPc-3) compared with human pancreatic duct cells (hTERT-HPNE). The densitometry read- ings/intensity ratio of each band and the original whole blot could be found in Table S9 and Figure S5.

Article Snippet: Three kinds of PDAC cell lines (MIA PaCa-2, SW1990, and BxPc-3) and immortalized human pancreatic duct cells (hTERT-HPNE) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Migration, In Vitro, Western Blot, Expressing

Journal: Cancers

Article Title: A LETM2-Regulated PI3K-Akt Signaling Axis Reveals a Prognostic and Therapeutic Target in Pancreatic Cancer.

doi: 10.3390/cancers14194722

Figure Lengend Snippet: Figure 5. Downregulation of LETM2 inhibits PDAC cell proliferation and metastasis and promoted cell apoptosis in vitro. (A) Western blotting analysis for verifying the efficiency of LETM2 knockdown. The densitometry readings/intensity ratio of each band and the original whole blot could be found in Table S11 and Figure S7. (B) Quantitative analysis of Western blotting and (C) qRT-PCR analysis revealed LETM2 protein and mRNA expression were significantly downregulated in knockdown cells. siLETM2 indicates BxPc-3 cells and SW1990 cells transfected with siRNA targeting LETM2. The relative quantification was calculated using the 2−∆∆Ct method and normalized based on GAPDH. (D) Cell apoptosis of LETM2 knockdown cell lines was measured using flow cytometry. (E) Cell growth of LETM2 knockdown cell lines was measured using CCK-8 assays. (F) Cell migratory and invasive ability of LETM2 knockdown cell lines were measured using transwell assay. Representative photographs (left) and quantification (right) were shown. Scale bars, 50 µm. The data are represented as mean ± SEM (n = 3 biologically independent experiments), and Student’s t-test was performed. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Three kinds of PDAC cell lines (MIA PaCa-2, SW1990, and BxPc-3) and immortalized human pancreatic duct cells (hTERT-HPNE) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vitro, Western Blot, Knockdown, Quantitative RT-PCR, Expressing, Transfection, Cytometry, CCK-8 Assay, Transwell Assay

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 1. Activin A is highly expressed in tumor specimens from PDAC patients and enters systemic circulation. (A) Gene expression analysis of INHBA in 18 tumor types of The Cancer Genome Atlas (TCGA) compared with corresponding matched normal (TCGA) tissues using RNA-Seq by Expectation–Maximization (RSEM). Sources for the statistics of cancer cachexia prevalence can be found in Supplemental References. (B) Representative immunofluorescence images of activin A (βA, AF488) and keratin 17 (CK17, AF568) expression in non-cancerous pancreatic tissue and PDAC tumor specimens. Nuclei were stained with DAPI. Scale bar = 75 µm. Corresponding serum activin A levels are indicated in ng/ml. Representative H&E images are displayed on the third row. Scale bar = 50 µm. (C) Area percentage of green pixels indicating activin A expression in immunofluorescence images from non-cancerous pancreatic tissue (n = 5) and PDAC tumor specimens (n = 6). Each dot represents the average green pixel area percentage per image as measured by Image J. (D) Measurement of activin A levels in sera of non-cancer patients (n = 25) and PDAC patients (n = 34). (E) Representative immunohistochemical images of activin A expression in pancreas tissue from healthy control as well as PanIN lesions and tumor specimens from KPC mice.

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

Techniques: Gene Expression, RNA Sequencing, Immunofluorescence, Expressing, Staining, Immunohistochemical staining, Control

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 2. Serum activin A levels can be correlated with loss of visceral adipose tissue in PDAC patients. (A) H&E images of visceral adipose tissue from PDAC patients with corresponding activin A levels in ng/ml. Scale bar = 150 µm. (B) Measurement of adipocyte diameter in visceral adipose tissue from PDAC patients. The diameters of adipocytes were measured from several histological sections. 6 sections, 0.761 ng/ml; 14 sections, 2.434 ng/ml; 14 sections, 3.966 ng/ml; 11 sections, 4.160 ng/ml; 10 sections, 7.459 ng/ml; 6 sections, 10.425 ng/ ml. Corresponding serum activin A levels are included in ng/ml. (C) Frequency curve of adipocyte diameter in visceral adipose tissue from patients with low serum activin A (0.761 ng/ml, red line), elevated serum activin A (8.984 ng/ml, blue line), and increasingly elevated serum activin A (10.425 ng/ml, black line). The diameters of adipocytes were measured with 6 histological sections each.

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

Techniques:

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 3. Activin A is highly expressed in tumor-derived cell lines from PDAC patients. (A) Immunofluorescence images of activin A (βA, top panels, AF488) and cytokeratin 19 (CK19, bottom panels, AF488) in hTERT-HPNE, PANC-1, BxPC-3 and MIA PaCa-2 cells. Nuclei were stained with DAPI. Scale bar = 75 µm. (B) Immunoblotting analysis of activin A expression in hTERT-HPNE, PANC-1, BxPC3, MIA PaCa-2 and HeLa cell lysates. (C) Relative INHBA mRNA expression levels in PANC-1 and BxPC-3 cells.

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

Techniques: Derivative Assay, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 4. Activin A is highly expressed in tumor specimens and tumor-derived cell lines of genetically engineered mouse models of PDAC. (A) Immunofluorescence images of activin A (AF488) in normal mouse pancreas tissue and tumor specimens of KPC 8060 and KPC 8069 mice. Scale bar = 50 µm. (B) Measurement of serum activin A levels in healthy control C567BL/6 J and KPC mice. (C) Immunoblotting analysis of activin A expression in healthy control C57BL/6 J mouse pancreas tissue and KPC8069 cell lysates. (D) Relative Inhba mRNA expression levels in healthy control C57BL/6 J mouse pancreas tissue and KPC8069 cells. (E) Measurement of activin A concentration in CM from KPC8060 and KCP8069 cells along with RPMI-1640. (F) Measurement of activin A concentration in CM from KPC8069 cells at 60% confluency, 100% confluency and 100% confluency following reintroduction (a blue bar) of fresh culture medium.

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

Techniques: Derivative Assay, Immunofluorescence, Control, Western Blot, Expressing, Concentration Assay

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 5. Activin A can be correlated with loss of gonadal visceral adipose tissue in an orthotopic implantation model of PDAC. (A) Representative immunohistochemical images of activin A expression in healthy control C57BL/6 J pancreatic tissue and tumor specimens of S2-013 mPDAC. Scale bar = 50 µm. (B) Measurement of serum activin A levels in sham and S2-013 mPDAC. (C) Mass of gonadal fat pad. (D) H&E images of adipose tissue from the gonadal fat pad in sham and S2-013 mPDAC. Scale bar = 150 µm. (E) Measurement of adipocyte diameter in gonadal adipose tissue from sham and S2-013 mPDAC. The diameters of adipocytes were measured from several histological sections (n = 5). The average adipocyte diameter from each animal was marked with a dot. F. Frequency curve of adipocyte diameter in gonadal fat pad of sham mice (dashed line) and S2-013 mPDAC (solid line).

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

Techniques: Immunohistochemical staining, Expressing, Control

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 7. Visceral adipose tissue remodeling in PDAC patients does not rely on increased browning of white adipose tissue. (A) Immunoblotting with UCP1 (33 kDa) in visceral adipose tissue of non-cancer patients (n = 5) and PDAC patients (n = 5) (top). Immuoblotting with UCP1 in visceral adipose tissue of a non-cancer patient and a PDAC patient with intrascapular brown adipose tissue from a healthy control mouse as a positive control (n = 2) (bottom). (B) Relative mRNA expression levels of UCP1 in visceral adipose tissue of non-cancer patients (n = 5) and PDAC patients (left). Corresponding serum activin A levels are included in ng/ml. Statistical analysis of relative mRNA expression levels of UCP1 in visceral adipose tissue of non-cancer patients (n = 5) and PDAC patients (n = 8) (right). n.s., not significant. (C) Representative immunofluorescence images of UCP1 expression in visceral adipose tissue from patients with low and high circulating activin A levels included in ng/ml. Nuclei were stained with DAPI. (D) Relative mRNA expression levels of HSL (LIPE) in visceral adipose tissue of non- cancer patients and PDAC patients (left). Corresponding serum activin A levels are included in ng/ml. Statistical analysis of relative mRNA expression levels of LIPE in adipose tissue of non-cancer patients (n = 4) and PDAC patients (n = 8) (right). n.s., not significant. Corresponding serum activin A levels are included in ng/ml.

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

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

Journal: Scientific reports

Article Title: Visceral adipose tissue remodeling in pancreatic ductal adenocarcinoma cachexia: the role of activin A signaling.

doi: 10.1038/s41598-022-05660-7

Figure Lengend Snippet: Figure 8. Fibrotic remodeling of visceral adipose tissue in PDAC patients and an orthotopic implantation model appears to correlate with activin A. (A) Picro-Sirius Red staining of gonadal adipose tissue of sham control and S2-013 mPDAC. Arrows indicate areas of collagen deposition. (B) Picro-Sirius Red staining of visceral adipose tissue in PDAC patients with corresponding activin A levels in ng/ml. Arrows indicate areas of collagen deposition. (C) Area percentage of red pixels indicating collagen I/III deposition in gonadal adipose tissue from sham control and S2-013 mPDAC as measured by Image J. The average area percentage of red pixels from each animal was marked with a dot. (D) Area percentage of red pixels indicating collagen I/III deposition in visceral adipose tissue from PDAC patients with corresponding activin A levels in ng/ml. The areas of collagen I/III deposition were measured from several histological sections. 7 sections, 0.761 ng/ml; 13 sections, 2.434 ng/ml; 14 sections, 3.966 ng/ml; 11 sections, 4.160 ng/ml; 7 sections, 8.984 ng/ml; 11 sections, 14.362 ng/ ml. Each dot indicates the area per section. (E) Immunofluorescence images with collagen I and fibronectin in visceral adipose tissue from patients with low and high serum activin A in ng/ml. Nuclei were stained with DAPI. Purple stars indicate adipocyte nuclei, while orange arrows indicate areas of signal. Scale bar = 50 µm for all microscope images. (F). Picro-Sirius Red staining of gonadal and subcutaneous white adipose tissue (WAT) obtained from C57BL/6 mice and cultured ex vivo with RPMI1640 (control), CM from KPC8069 cells, or CM from KPC8069 cells supplemented with FST288. Blue arrows indicate areas of collagen deposition.

Article Snippet: Human PDAC cell lines BxPC3 (CRL-1687, ATCC, Manassas, VA), PANC-1 (CRL-1469, ATCC), and MIA PaCa-2 (CRL-1420, ATCC); and human cervical adenocarcinoma cell line HeLa (CCL-2, ATCC) were maintained in RPMI-1640 (11875093, Gibco, Waltham, MA) or Dulbecco’s Modified Eagle Medium (DMEM, 12430054, Gibco) with 10% FBS (FBS001-HI, Neuromics Inc., Edina, MN) and 1% penicillin–streptomycin-glutamine (PSG) (10378016, Gibco).

Techniques: Staining, Control, Immunofluorescence, Microscopy, Cell Culture, Ex Vivo

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: High MUC1 expression in PDA cells positively correlates to TGF-βRII and negatively correlates to TGF-βRI levels. (A) The expression of MUC1-CT, TGF-βRI, TGF-βRII, and endogenous loading control β-actin in a panel of PDA cell lines, determined by Western blot. (B) Densitometric analysis of MUC1 expression versus TGF-βRI expression shows a negative correlation (Spearman’s correlation coefficient r = -0.2381, NS). (C) Densitometric analysis of MUC1 expression versus TGF-βRII expression shows a significantly positive correlation (Spearman’s correlation coefficient r = 0.8810, p = 0.0072).

Article Snippet: Human PDA cell lines (CFPAC, HPAC, HPAF-II, Panc1, MiaPaCa2, Su86.86 and BxPc3) were obtained from American Type Culture Collection and cultured as instructed.

Techniques: Expressing, Control, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: TGF-β exposure increases viability in cells with high MUC1 and reduces viability in low MUC1 PDA cells. MTT cell viability assay on (A) MiaPaca2.Neo cells with 10 ng/ml of TGF-β for 48 h. (B) HPAFII and (C) MiaPaca2. MUC1 cells with 10 ng/ml of TGF-β for 72 h. (D) HPAFII treated with control or MUC1 siRNA for 72 h followed by treatment with 10ng/ml of TGF-β for 24 h. All data are shown as means ± SEM of n = 3. Unpaired t-test was performed to compare between treated and untreated cells for each one of experiments A-C and two-way ANOVA was used to compare between untreated and treated in HPAFII.controlsiRNA and HPAFII.MUC1siRNA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human PDA cell lines (CFPAC, HPAC, HPAF-II, Panc1, MiaPaCa2, Su86.86 and BxPc3) were obtained from American Type Culture Collection and cultured as instructed.

Techniques: Viability Assay, Control