panc1 human pancreatic cell lines  (ATCC)

Journal: Frontiers in Pharmacology

Article Title: The human cathelicidin peptide LL-37 inhibits pancreatic cancer growth by suppressing autophagy and reprogramming of the tumor immune microenvironment

doi: 10.3389/fphar.2022.906625

Figure Lengend Snippet: LL-37 inhibits the growth of pancreatic cancer cells. (A) The cell viability of human PANC1 and MIA PaCa-2 cells treated with various concentrations of LL-37 (1–32 μM) for 24, 48 or 72 h was determined by MTT assay. (B) Colony formation assay was used to evaluate the proliferation ability of PANC1 and MIA PaCa-2 cells. (C) The proliferation of PANC1 and MIA PaCa-2 cells was detected by EdU staining (red). DAPI (blue) stained the nuclei. Cells treated with DMSO were used as negative controls. The data of all drug treatment groups were normalized to the control group. Data are presented as mean ± SD of three independent experiments conducted in triplicate. * p < 0.05, ** p < 0.01 vs . control (0 μM).

Article Snippet: The human pancreatic carcinoma cell lines PANC1 and MIA PaCa-2 isolated from PDAC cell origin as well as the BALB/c mice-derived macrophages RAW264.7 were purchased from American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: MTT Assay, Colony Assay, Staining, Control

Journal: Frontiers in Pharmacology

Article Title: The human cathelicidin peptide LL-37 inhibits pancreatic cancer growth by suppressing autophagy and reprogramming of the tumor immune microenvironment

doi: 10.3389/fphar.2022.906625

Figure Lengend Snippet: LL-37 induces DNA damage and cell cycle arrest in pancreatic cancer cells. LL-37 induced DNA damage by increasing γ-H2AX expression, as determined by immunofluorescence microscopy and western blotting (A,B ), respectively after LL-37 (8 and 16 μM) treatment for 24 h. (C) The cell cycle distributions in PANC1 and MIA PaCa-2 cells were analyzed by flow cytometry after LL-37 treatment for 24 h. The data are presented as means ± SD. * p < 0.05, ** p < 0.01 vs . control (0 μM) value in the G2 phase. (D) The expressions of cell cycle arrest related proteins were determined by western blotting after LL-37 treatment for 24 h. Non-arbitrary band density data relative to the housekeeping protein ß -actin are presented as mean ± SD, * p < 0.05, ** p < 0.01 vs. control (0 μM).

Article Snippet: The human pancreatic carcinoma cell lines PANC1 and MIA PaCa-2 isolated from PDAC cell origin as well as the BALB/c mice-derived macrophages RAW264.7 were purchased from American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Expressing, Immunofluorescence, Microscopy, Western Blot, Flow Cytometry, Control

Journal: Frontiers in Pharmacology

Article Title: The human cathelicidin peptide LL-37 inhibits pancreatic cancer growth by suppressing autophagy and reprogramming of the tumor immune microenvironment

doi: 10.3389/fphar.2022.906625

Figure Lengend Snippet: LL-37 inhibits autophagy in pancreatic cancer cells. (A) LL-37 (8 and 16 μM) decreased the formation of autophagosomes in pancreatic cancer cells as determined using immunofluorescence microscopy. (B) Autophagic vacuoles in PANC1 and MIA PaCa-2 cells treated with 16 μM LL-37 or DMSO were observed by TEM. The red arrow indicates autophagic vacuoles. (C) The autophagic flux in LL-37-treated PANC1 and MIA PaCa-2 cells was tested using the Autophagy Tandem Sensor RFP-GFP-LC3B. Representative images were photographed under a confocal microscope. The average numbers of yellow and red LC3B dots per cell are shown, **p < 0.01 vs. control of yellow dots, ##p < 0.01 vs . control of red dots. (D) The levels of autophagy markers LC3II, Atg5–Atg12 and Beclin-1 in PANC1 and MIA PaCa-2 cells were measured by western blotting after treatment with LL-37 for 24 h. (E) The mTOR pathway involved in the inhibitory effect of LL-37 on autophagy was determined by western bloting. Cells treated with DMSO were used as negative controls. Three independent experiments were each performed in triplicate. Representative protein bands are shown for comparison. Non-arbitrary band density data relative to the housekeeping protein ß -actin are presented as mean ± SD, * p < 0.05, ** p < 0.01 vs . control (0 μM).

Article Snippet: The human pancreatic carcinoma cell lines PANC1 and MIA PaCa-2 isolated from PDAC cell origin as well as the BALB/c mice-derived macrophages RAW264.7 were purchased from American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Immunofluorescence, Microscopy, Control, Western Blot, Comparison

Journal: Frontiers in Pharmacology

Article Title: The human cathelicidin peptide LL-37 inhibits pancreatic cancer growth by suppressing autophagy and reprogramming of the tumor immune microenvironment

doi: 10.3389/fphar.2022.906625

Figure Lengend Snippet: LL-37 increases the intracellular production of reactive oxygen species (ROS) in pancreatic cancer cells. (A) ROS were detected using the fluorescence dye CM-H2DCFDA and immunofluorescence microscopy. ROS fluorescence was measured using a SpectraMax Plate Reader (Molecular Devices) with excitation/emission wavelengths of 495/520 nm. Data are presented as mean ± SD of three independent experiments conducted in triplicate. * p < 0.05, ** p < 0.01 vs . control (0 μM). (B) Effect of LL-37 on the MMP in PANC1 and MIA PaCa-2 cells was measured using JC-1 staining method. Representative images were photographed under a confocal microscope. The red/green fluorescence ratio was quantified by the Image J. ** p < 0.01 vs. control (0 μM). (C) The ROS production of pancreatic cancer cells treated with LL-37 in the presence and absence of a ROS scavenger NAC (2 mM) was determined. * p < 0.05 vs . control; # p < 0.05 vs . LL-37 (8 µM). (D) The viability of pancreatic cancer cells treated with LL-37 in the presence and absence of a ROS scavenger NAC was determined using an MTT assay. ** p < 0.01 vs . control; # p < 0.05, ## p < 0.01 vs . LL-37 (8 µM). (E) The expression of γ-H2AX in PANC1 and MIA PaCa-2 cells treated with LL-37 in the presence and absence of NAC was determined by western blotting. Non-arbitrary band density data relative to that of the housekeeping protein ß -actin are presented as mean ± SD, * p < 0.05, ** p < 0.01 vs . LL-37 (8 µM). The ROS production (F) and cell viability (G) were determined in pancreatic cancer cells treated with LL-37 in the presence and absence of an autophagy inducer (rapamycin; 100 nM), respectively. * p < 0.05, ** p < 0.01 vs . control; # p < 0.05 vs. LL-37 (8 µM).

Article Snippet: The human pancreatic carcinoma cell lines PANC1 and MIA PaCa-2 isolated from PDAC cell origin as well as the BALB/c mice-derived macrophages RAW264.7 were purchased from American Type Culture Collection (ATCC, Manassas, VA, United States).

Techniques: Fluorescence, Immunofluorescence, Microscopy, Control, Staining, MTT Assay, Expressing, Western Blot