Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a-e, The indicated KRAS G12D -mutant cells ( a , c , e ) and organoids ( b , d ) were treated with increasing concentration of MRTX1133 (MRTX) or HRS4642 (HRS) for 72 h (cells) or 7 days (organoids). Total viable cells are presented relative to vehicle (Veh). IC 50 values are provided in Source Data. Parental HPAC, AsPC-1, SW1990, and No.6 cells are denoted as KS HPAC, AsPC-1, SW1990, and No.6, respectively. Acquired KRASi resistant HPAC, AsPC-1, SW1990, No.6 cells are labeled as AR. f - i , IB analysis of indicated proteins in human PDAC cells treated -/+ 100 nM MRTX for 2 h ( f , g ), or -/+ HRS at the indicated concentrations for 2 h ( h , i ). j , k , IB analysis of indicated proteins in human PDAC organoids treated -/+ 100 nM MRTX or HRS for 24 h. l , Viability of WT and KRAS-ablated (KRAS Δ ) KS and KR/AR cells is presented relative to day 0. IB analysis of KRAS in the same cells is shown below. Data in ( a - e and l) (n=3 independent experiments) are mean ± s.e.m. Statistical significance was determined using one-way ANOVA with Tukey post-hoc tests (SW1990, HPAC, and AsPC-1 in l ) or two-sided unpaired t-test (KP-4, PANC-1 in l ) based on data normality distribution. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant. Exact P values are shown in Source Data.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: Mutagenesis, Concentration Assay, Labeling

Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a - c , RNA-seq analysis was performed on KS (AsPC1) and KR (PANC-1) PDAC cells treated -/+ MRTX for 24 h. ( a ) Venn diagram showing the overlap of differentially expressed genes between MRTX- and Veh-treated cells and between MRTX-treated AsPC1 and MRTX-treated PANC-1 cells. ( b ) KEGG enrichment analysis of selected overlapping genes (n = 1,339 genes) from ( a ), representing genes whose expression is specifically altered in intrinsic resistant cells upon MRTX treatment. ( c ) Heatmap showing mRNA expression of lipid metabolism-related genes. Blue, replicates with low expression; red, replicates with high expression. d , e , qPCR analysis of indicated mRNAs in the indicated cell lines or organoids treated -/+ MRTX for 24 h. f , LC-MS quantification of intracellular acyl-coenzyme A levels in PANC-1 cells treated -/+ MRTX for 24 h. g , Fractional labelling of TCA cycle intermediates in PANC-1 cells cultured with [U- 13 C]-PA, -/+ MRTX for 12 h. h , OCR of KS and AR AsPC-1 cells treated -/+ MRTX for 24 h before and after treatment with Omy, FCCP, and rotenone/antimycin A (ROT/AA). i , Total cellular ATP in indicated KS and KR/AR cells treated -/+ MRTX for 48 h. Data are presented relative to untreated cells. KP-4 is intrinsically KRASi resistant (KR). j , OCR of KR PANC-1 cells expressing DOX ACSBG1Δ treated -/+ DOX for 48h, followed with -/+ MRTX for 24 h before and after treatment with Omy, FCCP, and rotenone/antimycin A. k , Total cellular ATP in the indicated cells treated -/+ DOX for 48 h, followed with -/+ MRTX or HRS for 48 h. Data are presented relative to untreated cells. l , IB analysis of the indicated proteins in the indicated cells treated -/+ DOX for 48 h. Data in ( f , h - k ) (n=3 independent experiments) are mean ± s.e.m. Statistical significance was determined using two-sided unpaired t -test ( f , i KP-4) or one-way ANOVA with Tukey post-hoc tests ( i , k ) based on data normality distribution. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant. Exact P values are shown in Source Data.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: RNA Sequencing, Expressing, Liquid Chromatography with Mass Spectroscopy, Cell Culture

Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a , LC-MS analysis of intracellular acyl-coenzyme A species in sensitive (KS) and acquired (AR) AsPC-1 cells -/+ 100 nM MRTX1133 (MRTX) for 24 h. Veh, Vehicle. b , Right: Fractional labelling of TCA cycle intermediates in KS/AR AsPC-1 cells incubated with [U- 13 C]-PA and -/+ MRTX for 12 h. α-KG, α-ketoglutarate. Left: A schematic illustration of PA-derived acetyl-CoA fuels the TCA cycle. Blue, replicates with low expression; red, replicates with high expression. c , Oxygen consumption rate (OCR) of KS and AR AsPC-1 cells incubated -/+ PA and 100 nM HRS-4642 (HRS) for 24 h before and after addition of 40 μM Etomoxir (Eto), followed by oligomycin (Omy), FCCP, and rotenone/antimycin A (ROT/AA) treatments. d , KS/AR HPAC cells expressing doxycycline (DOX)-inducible CPT1 deletion (DOX CPT1Δ ) were treated -/+ 0.2 μg/mL DOX for 48 h, and then -/+ HRS for 48 h. Total cellular ATP is relative to untreated KS cells. e , Representative images of mitochondria (TIM23) and BODIPY-labelled lipid droplets (LD) in parental KS No.6 and AR No.6 human PDAC organoids treated -/+ MRTX for 24 h. f , Quantification of mitochondria, LD, and their co-localization in ( e ). g , Pancreas morphology 3 weeks after orthotopic transplantation of KS and AR KC6141 (KC) cells expressing DOX CPT1Δ and host treatment at 72 h post-transplantation -/+ 30 mg/kg MRTX (i.p.), 0.2 mg/mL DOX (drinking water), or DOX + MRTX. h , Pancreas weight relative to body weight (P/B weight) of mice in ( g ). i , Representative H&E and IHC staining for CK19 in pancreata from ( g ). Boxed areas are further magnified. Tumor areas are at the bottom. j , Representative AFADESI MSI images and quantification of endogenous C16:0 L-carnitine (m/z 400.3436) and C18:1 L-carnitine (m/z 426.3559) distribution in pancreata two weeks after orthotopic transplantation of KS/AR KC cells expressing DOX CPT1Δ . Host were treated starting 10 days post-transplantation with MRTX, DOX, or DOX + MRTX. AR tumors exhibit elevated FAO and increased sensitivity to CPT1 ablation compared to KS tumors. Boxed regions indicate tumor areas. k , Pancreata from mice in ( j ) were sectioned and cultured with 100 μM [U- 13 C]-PA for 4 h. Representative MALDI MSI images and quantification of unlabeled citrate (M + 0, m/z 191.019) and 13 C-citrate (M + 2, m/z 193.026) distribution in pancreata, showing a similar trend to ( j ). Data in ( a , c , d , k ) (n=3 independent experiments), ( f ) (n=25 fields), ( h ) (n = 5 mice), ( i ) (n=7 fields), and ( j ) (n=6 fields) are mean ± s.e.m. Statistical significance was determined using one-way analysis of variance (ANOVA) with Tukey post-hoc tests ( a , d , f , h - k ) based on data normality distribution. Exact P values are shown in the Source Data. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant. Scale bars ( e ) 40 μm, ( g ) 1 cm, ( i ) 100 μm, ( j , k ) 500 μm.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: Liquid Chromatography with Mass Spectroscopy, Incubation, Derivative Assay, Expressing, Transplantation Assay, Immunohistochemistry, Cell Culture

Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a , b , Genes differentially expressed between KS (AsPC-1, SW1990) and KR/AR (PANC-1, SW1990) cells treated -/+ MRTX for 24 h. Blue, replicates with low expression; red, replicates with high expression. c , d , IB analysis of ADGRB1 in indicated organoids or cell lines treated -/+ HRS for 24 h ( c ) or -/+ DOX for 48 h ( d ). e , KS and AR cells expressing DOX ADGRB1Δ , SRF-responsive luciferase reporter (SRF-RE), and a Renilla-luciferase control were treated -/+ DOX for 48 h, followed by incubation with D-Luciferin. Normalized Firefly/Renilla luciferase ratios, reflecting ADGRB1 (Gα12/13) activity, are shown relative to untreated KS cells. f , Fractional labelling of TCA cycle intermediates in KS and AR HPAC cells expressing DOX ADGRB1Δ treated -/+ DOX for 48 h, and then incubated with [U- 13 C]-PA and MRTX for 12 h. Blue, replicates with low expression; red, replicates with high expression. g , Representative IHC of resected ADGRB1 hi (#556) and ADGRB1 lo (# 835) human PDAC tissues. Boxed areas are further magnified. h , Numbers of human PDAC specimens (n = 42) positive for the indicated proteins, indicated as low and high expression. i , Correlation between expression levels of the indicated proteins in human specimens examined by a two-tailed Chi-square test. j , Comparisons of overall survival between patients with PDAC stratified according to ADGRB1 and pS144-PAK1. Significance was determined by log-rank test. Data in ( e ) (n=3 independent experiments) are mean ± s.e.m. Statistical significance was determined using one-way ANOVA with Tukey post-hoc tests ( e ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Exact P values are shown in Source Data.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: Expressing, Luciferase, Control, Incubation, Activity Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a , Immunoblot (IB) analysis of indicated proteins in acquired KRASi-resistant SW1990 cells (AR) -/+ knock down (KD) of indicated GPCRs and -/+ MRTX for 2 h. b , AR and KS KC cells expressing DOX ADGRB1Δ were orthotopically transplanted into mice -/+ 7 days MRTX, DOX, or DOX + MRTX, initiated 72 h post-transplantation. Then the BODIPY signal in PDAC cells was analyzed by flow cytometry 24 h after i.p. of BODIPY-PA loaded adipocytes (3T3-L1). The experimental scheme is shown to the left. c , IB analysis of indicated proteins in the indicated KS and AR AsPC-1 cells -/+ Flag-tagged PI3Kγ R1021C expression and -/+ DOX treatment for 48 h, followed by -/+ MRTX for 2 h. d , Representative images and quantification of MP in TMR-DEX-incubated cells from ( c ). e , OCR of cells in ( c ) treated -/+ DOX for 48 h, followed by -/+ MRTX or EIPA for 24 h before and after -/+ PA incubation, and treatment with Omy, FCCP, and rotenone/antimycin A. f , Representative H&E and IHC staining of resected human PDAC tissues classified as KS or KR based on organoid sensitivity to KRASi . Boxed areas are further magnified. g , Correlations between the indicated proteins and KRASi sensitivity in ( f ) were analyzed by a two-tailed Chi-square test. Data in b (n=3 mice), d (n=20 fields) and e (n=3 independent experiments) are mean ± s.e.m. Statistical significance was determined using Brown-Forsythe and Welch ANOVA tests with Dunnett T3 test ( d ) based on data normality distribution. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Exact P values are shown in Source Data. Scale bars ( d ) 10 μm, ( g ) 100 μm.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: Western Blot, Knockdown, Expressing, Transplantation Assay, Flow Cytometry, Incubation, Immunohistochemistry, Two Tailed Test

Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a , IB analysis of indicated proteins in KRAS G12C mutated KS and AR MIA PaCa-2 cells (MIA) or KRAS G12D mutated GP2D cells -/+ 100 nM of the KRAS G12C i AMG510 or the KRAS G12D i HRS for 24 h. b , Representative images and quantification of MP in KS/AR GP2D cells expressing DOX ADGRB1Δ treated -/+ DOX for 48h, followed by treatment -/+ 1 μM PI3Kγi IPI549 (IPI), 1 μM PI3Kαi LY494002 (LY), HRS, HRS + IPI, or HRS + LY for 24 h. c , OCR of indicated cells -/+ DOX for 48h, followed by -/+ AMG510 (MIA) or HRS (GP2D) for 24 h, before and after -/+ PA, followed by Omy, FCCP, and rotenone/antimycin A treatments. d , e , Total cellular ATP in KS and AR MIA PaCa-2 or GP2D cells expressing DOX ADGRB1Δ treated with the indicated compounds for 48h. f , Representative IHC and quantification in resected human colon adenocarcinoma tissues before and after KRAS G12D i QLC1101 treatment. Data in ( b ) (n=15 fields), ( c , d , e ) (n=3 independent experiments), and ( f ) (n=6 fields) are mean ± s.e.m. Statistical significance was determined using one-way ANOVA with Brown-Forsythe and Welch corrections followed by Dunnett T3 test ( b ), or with Tukey post-hoc tests ( d , e ), or two-sided unpaired t -test or Mann-Whitney U -test ( f ) based on data normality distribution. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Exact P values are shown in Source Data. Scale bars ( b ) 20 μm, ( f ) 100 μm.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: Expressing, MANN-WHITNEY

Journal: bioRxiv

Article Title: Fatty acid scavenging enables cancer escape from KRAS inhibition

doi: 10.64898/2026.04.01.715565

Figure Lengend Snippet: a , KS/AR No.6 organoids were treated with IPI and/or HRS at the indicated concentrations for 7 days, and cell viability was assessed with CellTiter-Glo (CTG). Upper: Dose-response matrix (viability) for IPI and HRS. Blue intensity indicates the degree of inhibition. Lower: 3D synergy plots generated by SynergyFinder + . The synergy score is shown by red (>0) and green (<0). The synergy score > 10 indicates strong synergic effect. b , Akaluciferase bioluminescence imaging of mice 3 weeks after orthotopic transplantation of KS or AR KC cells expressing DOX ADGRB1Δ and a Ki67-akaluciferase reporter, -/+ MRTX, DOX, or DOX + MRTX treatments initiated 72 h post-transplantation. Quantification of akaluciferase activity, expressed as relative light units (RLU), is on the right. c , P/B ratio of mice from ( b ). d , Representative H&E and CK19 IHC staining of pancreata from ( b ). Boxed areas are further magnified. e , Quantification of tumor areas from ( d ). f , Pancreas morphology and P/B ratio 3 weeks after orthotopic transplantation of indicated cells into mice -/+ MRTX, 15 mg/kg IPI (p.o.), or MRTX + IPI treatments initiated 72 h post-transplantation. g , Representative H&E and CK19 staining of pancreata from ( f ). Boxed areas are further magnified. Quantification of tumor areas is shown to the right. h , Flow cytometry analysis of BODIPY signals in PDAC cells harvested 24 h after i.p. injection of BODIPY-PA-loaded adipocytes into mice from ( f ) at day 7 post-KS and AR KC cell implantation. Data in ( b ) (n=5 mice), ( c , f ) (n=5-7 mice), ( e , g ) (n=6 fields), and ( h ) (n=3 mice) are mean ± s.e.m. Statistical significance was determined using one-way ANOVA with Tukey post-hoc tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Exact P values are shown in Source Data. Scale bars ( d , g ) 100 μm, ( f ) 1 cm.

Article Snippet: A total of 16 specimens of human PDAC bearing KRAS G12D mutation were acquired from patients who were diagnosed with PDAC between July 2023 and September 2025 at The First Affiliated Hospital of Anhui Medical University.

Techniques: Inhibition, Generated, Imaging, Transplantation Assay, Expressing, Activity Assay, Immunohistochemistry, Staining, Flow Cytometry, Injection

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: Antitumor activity of BiTP combinatorial chemotherapy in orthotopic PDAC models. (A) Treatment plan and schedules. (B) Schematic diagram of the KPC model. The approximate time frame of mouse tumor progression to borderline resectable (BR), locally advanced (LA), and terminal (TERM) are days 7, 14, and 21, respectively. (C) Representative tumor images of orthotopic PDAC mice receiving different therapies. (D) Tumor weights and volumes on day 30. (E) The table included the survival rate (%), tumor proliferation (T/C, %), tumor growth inhibition (TGI, %), and resectability status of orthotopic PDAC mice receiving different therapies on day 30, and the median survival time (MST) in survival assays. (F) Kaplan–Meier plot survival curve and pairwise comparison results of orthotopic PDAC mice receiving different therapies in survival assays. Data presented in the graphs represent mean±standard deviation (SD) (**** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05; ns P >0.05). BiTP, anti-PD-L1/TGF-β antibody; CTH, chemotherapy; PDAC, pancreatic ductal adenocarcinoma; PD-L1, programmed cell death 1 ligand 1.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: Activity Assay, Inhibition, Comparison, Standard Deviation

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: Decisions about resectability status in PDAC murine models.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: Membrane

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: Preclinical assessment of neoadjuvant BiTP combinatorial chemotherapy for the treatment of PDAC. (A) Treatment plan and schematic diagram of treatment schedules. (B) Schematic diagram of distal pancreatectomy in orthotopic PDAC mice model. (C) Representative images of tumors. (D) Tumor weights and volumes of direct surgery (day 7) and post-neoadjuvant therapy resections (day 24). (E) Table of resectability status (day 24), operative mortality (OM), and median survival time (MST) of orthotopic PDAC mice receiving different therapies. (F–H) Kaplan–Meier plot survival curve and pairwise comparison results of orthotopic PDAC mice receiving different therapies in survival assays. Data presented in the graphs represent mean±standard deviation (SD). (**** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05; ns P >0.05). AC, adjuvant chemotherapy; ACI, adjuvant chemo-immunotherapy; BiTP, anti-PD-L1/TGF-β antibody; CTH, chemotherapy; NAC, neo-adjuvant chemotherapy; NACI, neo-adjuvant chemo-immunotherapy; NAT, neo-adjuvant therapy; SRG, surgery.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: Comparison, Standard Deviation, Adjuvant

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: BiTP reverses TGF-β-induced EMT (epithelial–mesenchymal transition) of PDAC (pancreatic ductal adenocarcinoma) cells. (A) CCK-8 (Cell Counting Kit-8) assay to measure the effect of BiTP on TGF-β-mediated chemoresistance in PDAC cells. After 10 ng/ml TGF-β1, 10 5 pM BiTP or hIgG treatment for 24 h. HPDE6-C7, CFPAC-1, ASPC-1, and KPC cells growing in 96-well plates were exposed to serial dilutions of gemcitabine and nab-paclitaxel for 72 h and CCK-8 assays was performed. (B) The representative images of IHC (immunohistochemistry) staining of Ki67 in orthotopic PDAC mice model and statistical graph of the percentage of Ki67-positive cells. (C) The representative images of TUNEL staining for apoptosis (green) and nuclei (DAPI, blue) in orthotopic PDAC mice model and statistical graph of the percentage of apoptosis cells. (D) Transwell assays to measure the effect of BiTP on TGF-β-mediated migration and in PDAC cells. After 10 ng/ml TGF-β1, 10 5 pM BiTP or hIgG treatment for 96 h. 1×10 4 CFPAC-1 and KPC cells were seeded in the upper chambers and Transwell assays were performed. (E) Western blotting assays exploring the chemotherapy-induced EMT and the blocking effect of BiTP. (F) The representative images of IHC staining of E-cadherin in orthotopic PDAC mice model. Statistical graph of the AOD (average optical density) of E-cadherin. (G) The representative images of IF staining of Vimentin (green) and α-SMA (rose red) in orthotopic PDAC mice model. Statistical graph of the arbitrary units of Vimentin and α-SMA. (H) The representative images of Masson staining in orthotopic PDAC mice model. Statistical graph of the percentage of collagen volume fraction (%). Data presented in the graphs represent mean±standard deviation (SD). (**** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05; ns P >0.05). BiTP, anti-PD-L1/TGF-β antibody; CTH, chemotherapy; KPC, LSL-Kras(+/G12D);LSL-Trp53(+/R172H);Pdx1-Cre; PDAC, pancreatic ductal adenocarcinoma; PD-L1, programmed cell death 1 ligand 1; TGF-β, transforming growth factor-β.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: CCK-8 Assay, Cell Counting, Immunohistochemistry, Staining, TUNEL Assay, Migration, Western Blot, Blocking Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Role of miR-30a-3p Regulation of Oncogenic Targets in Pancreatic Ductal Adenocarcinoma Pathogenesis

doi: 10.3390/ijms21186459

Figure Lengend Snippet: Tumor-suppressive functions of miR-30a-5p and miR-30a-3p in pancreatic ductal adenocarcinoma (PDAC) cells. ( A ) Expression levels of miR-30a-5p and miR-30a-3p in PDAC clinical specimens and cell lines (PANC-1 and SW1990). Data were normalized relative to the expression of RNU48. ( B ) Pearson’s coefficient showed positive correlations between the expression levels of miR-30a-5p and miR-30a-3p in clinical specimens. ( C ) Cell proliferation assessed using XTT assays. Data were collected 72 h after miRNA transfection (* p < 0.0001). ( D ) Cell migration assessed with a membrane culture system. Data were collected 48 h after seeding the cells into the chambers (* p < 0.0001). ( E ) Cell invasion determined using Matrigel invasion assays conducted 48 h after the seeding of the miRNA-transfected cells into the chambers (* p < 0.0001).

Article Snippet: PDAC cell lines SW1990 and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA, USA) and RIKEN Cell Bank (Tsukuba, Ibaraki, Japan), respectively.

Techniques: Expressing, Transfection, Migration, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Role of miR-30a-3p Regulation of Oncogenic Targets in Pancreatic Ductal Adenocarcinoma Pathogenesis

doi: 10.3390/ijms21186459

Figure Lengend Snippet: Identification of putative targets regulated by miR-30a-3p in PDAC cells.

Article Snippet: PDAC cell lines SW1990 and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA, USA) and RIKEN Cell Bank (Tsukuba, Ibaraki, Japan), respectively.

Techniques: Expressing, Ubiquitin Proteomics, Virus, Binding Assay, Dominant Negative Mutation, Sequencing, Coagulation, Immunopeptidomics, RNA Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Role of miR-30a-3p Regulation of Oncogenic Targets in Pancreatic Ductal Adenocarcinoma Pathogenesis

doi: 10.3390/ijms21186459

Figure Lengend Snippet: GSE15471 dataset analyses of the expression levels of 12 genes (predicted 5-year survival) that are targets for miR-30a-3p regulation in PDAC clinical specimens.

Article Snippet: PDAC cell lines SW1990 and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA, USA) and RIKEN Cell Bank (Tsukuba, Ibaraki, Japan), respectively.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Role of miR-30a-3p Regulation of Oncogenic Targets in Pancreatic Ductal Adenocarcinoma Pathogenesis

doi: 10.3390/ijms21186459

Figure Lengend Snippet: Effects of ITGA2 knockdown in SW1990 cells. ( A ) mRNA and ( B ) protein expression levels of ITGA2 by transfection with two types of si ITGA2 in SW1990 cells. ( C,D,E ) Cell proliferation, migration and invasion assays in SW1990 cells. (* p < 0.0001)

Article Snippet: PDAC cell lines SW1990 and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA, USA) and RIKEN Cell Bank (Tsukuba, Ibaraki, Japan), respectively.

Techniques: Knockdown, Expressing, Transfection, Migration

Journal: International Journal of Molecular Sciences

Article Title: Role of miR-30a-3p Regulation of Oncogenic Targets in Pancreatic Ductal Adenocarcinoma Pathogenesis

doi: 10.3390/ijms21186459

Figure Lengend Snippet: ITGA2 overexpression in clinical PDAC specimens. Representative immunohistochemical images with staining of ITGA2 and ITGB1 in clinical samples. Overexpression of ITGA2 and ITGB1 was detected in the cancer lesions. Images of samples from Patient ( A ) number 20 and ( B ) number 17 are shown.

Article Snippet: PDAC cell lines SW1990 and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA, USA) and RIKEN Cell Bank (Tsukuba, Ibaraki, Japan), respectively.

Techniques: Over Expression, Immunohistochemical staining, Staining

Journal: Bioengineering & Translational Medicine

Article Title: Adjustable extracellular matrix rigidity tumor model for studying stiffness dependent pancreatic ductal adenocarcinomas progression and tumor immunosuppression

doi: 10.1002/btm2.10518

Figure Lengend Snippet: Matrix stiffness is related to the prognosis of PDAC. (a) Representative images of PDAC cancerous tissue and paired para‐cancer tissues stained with Masson trichrome (Masson); (b and c) Paired dot plots and bar graphs presented the PDAC cancerous tissue and paired para‐cancer tissues percentile values for collagen volume fraction; (d) Representative images of Masson‐stained PDAC cancerous tissue at different stages of AJCC 8th tumor stages (I, II, III, IV); (e) Bar graphs illustrated the percentile values of PDAC cancerous tissue for collagen volume fraction in different stages of AJCC 8th tumor stages (I, II, III, IV); (f) Bar graphs displayed the PDAC cancerous tissue percentile values for collagen volume fraction in different stages of the AJCC 8th TNM tumor stages; (g) Kaplan–Meier survival curve for the PDAC patients with different cancer matrix stiffness; (h) Bar graphs illustrated the PDAC patients with different stiffness of the cancer matrix.

Article Snippet: The human PDAC cell lines PAN1 (with Kras G12D mutation) and CFPAC‐1 (with Kras G12V mutation), and BXPC3 (without Kras mutation), were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: Staining

Journal: Bioengineering & Translational Medicine

Article Title: Adjustable extracellular matrix rigidity tumor model for studying stiffness dependent pancreatic ductal adenocarcinomas progression and tumor immunosuppression

doi: 10.1002/btm2.10518

Figure Lengend Snippet: The influence of matrix stiffness on cell morphology, proliferation, and migration of PDAC cells in vitro. (a) PDAC cells encapsulated in 3D hydrogels with different matrix stiffness (1, 10, and 20 kPa); (b) Cells cultured for 5 days under 3D conditions with different matrix stiffness, cell size varies with matrix stiffness and incubation time, showing a significant gradient of change; (d and e) Increasing matrix stiffness also resulted in thickening of cytoskeletal organization stress fibrils and increased actin and α‐tubulin expression (actin staining as green and α‐tubulin staining as red); (f) The clone‐forming assay shows that changes in matrix stiffness affect cell clone‐forming abilities; (g) Schematic of a 3D cell culture system based on cross‐linked GelMA hydrogels with visible light (405 nm) cross‐linked GelMA hydrogels. Data presented in the graphs represent means ± SD. *** p < 0.001; ** p < 0.01; * p < 0.05; ns p > 0.05.

Article Snippet: The human PDAC cell lines PAN1 (with Kras G12D mutation) and CFPAC‐1 (with Kras G12V mutation), and BXPC3 (without Kras mutation), were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: Migration, In Vitro, Cell Culture, Incubation, Expressing, Staining

Journal: Bioengineering & Translational Medicine

Article Title: Adjustable extracellular matrix rigidity tumor model for studying stiffness dependent pancreatic ductal adenocarcinomas progression and tumor immunosuppression

doi: 10.1002/btm2.10518

Figure Lengend Snippet: Adjustable extracellular matrix rigidity orthotopic tumor transplantation murine models of PDAC the influence of matrix stiffness in vivo. (a) Schematic representation of our established methods for preparing GelMA‐based adjustable extracellular matrix rigidity orthotopic tumor transplantation murine models; (b) Change in body weight of the mice in different groups; (c and d) Representative pictures of orthotopic tumors excised from mice; (e) Bar graph showing the average tumor volume and weight, respectively, of mice with different matrix stiffness PDAC tumors; (f) Representative pictures of greyscale and shear wave elastography ultrasound images of GelMA‐based adjustable extracellular matrix rigidity orthotopic tumor determined by noninvasive ultrasound evaluation. Side‐by‐side display of anatomical 8‐mode US image (gray image, top) and an elastography image restricted to the set study box (colored image, button) obtained with echographic 8‐mode and 2D‐SWE mode, coupled to doppler mode. The red color represents stiff tissue and the blue color reflects soft tissue (scale 0–50 kPa); (g) Bar graph showing the average tumor elastic modulus, respectively, of the orthotopic tumor of mice with different matrix stiffness PDAC tumors. Data presented in the graphs represented means ± SD. *** p < 0.001; ** p < 0.01; * p < 0.05; ns p > 0.05.

Article Snippet: The human PDAC cell lines PAN1 (with Kras G12D mutation) and CFPAC‐1 (with Kras G12V mutation), and BXPC3 (without Kras mutation), were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: Transplantation Assay, In Vivo, Shear

Journal: Bioengineering & Translational Medicine

Article Title: Adjustable extracellular matrix rigidity tumor model for studying stiffness dependent pancreatic ductal adenocarcinomas progression and tumor immunosuppression

doi: 10.1002/btm2.10518

Figure Lengend Snippet: Bulk RNA‐seq and space transcriptome to the mechanisms of stiffness‐dependent PDAC progression. Adjustable orthotopic extracellular matrix rigidity tumor transplantation murine models of PDAC were built in immune‐competent C57BL/6J mice. After 2 weeks, tumor tissues were collected for bulk RNA‐seq assay. (a) Bar plot showing KEGG enrichment analysis of the DEPs (upregulated and downregulated); (b) Bar plot showing GO enrichment analysis of the DEPs (upregulated and downregulated); (c) The heat maps representing the GSVA analysis based on Hallmark gene sets; (d and e) Space transcriptome mapping immune infiltration in the pancreatic ductal adenocarcinoma microenvironment using public data. Including original tissue images stained with HE (marked by the pathologist and showing the cancer foci and stroma), the cancer foci and stroma shown by spatial transcriptome analysis, and the spatial distribution of the various cell types (red shows enrichment) mapped based on spatial transcriptome data analysis.

Article Snippet: The human PDAC cell lines PAN1 (with Kras G12D mutation) and CFPAC‐1 (with Kras G12V mutation), and BXPC3 (without Kras mutation), were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: RNA Sequencing, Transplantation Assay, Staining

Journal: Bioengineering & Translational Medicine

Article Title: Adjustable extracellular matrix rigidity tumor model for studying stiffness dependent pancreatic ductal adenocarcinomas progression and tumor immunosuppression

doi: 10.1002/btm2.10518

Figure Lengend Snippet: Extracellular matrix rigidity promotes EMT and the tumor immunosuppression microenvironment. To further confirm the findings from the bioinformatics analysis, we validated protein levels and difference in immune cell infiltration in vitro and in vivo. (a) Immunoblotting analysis of EMT (E‐cadherin, Vimentin), TGFβ1, and PDL1 expression in PDAC cells cultured in different matrix rigidity 3D culture systems (1–20 Kpa); (b) Multiplex IHC staining to assess collagen production level (Masson staining), EMT status (E‐cadherin) and cell proliferation (Ki‐67) in different matrix rigidity murine PDAC tumor tissues; (c) Multiplex IF staining to map the TME of in different matrix rigidity murine PDAC tumors tissues include CD8 + T cells (CD8 + ), Exhaust CD8 + cytotoxic T (CD8 + PD1 + ), CD4 + T cells (CD4 + ), regulatory T cells (Treg), macrophages (F4/80 + ), M2 macrophages (F4/80 + CD206 + ), EMT status (Vimentin and α‐SMA), and TGFβ1/PDL1 expression.

Article Snippet: The human PDAC cell lines PAN1 (with Kras G12D mutation) and CFPAC‐1 (with Kras G12V mutation), and BXPC3 (without Kras mutation), were obtained from the China Center for Type Culture Collection (CCTCC).

Techniques: In Vitro, In Vivo, Western Blot, Expressing, Cell Culture, Multiplex Assay, Immunohistochemistry, Staining