Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 4 ch5E6 treatment leads to a substantial decrease in the growth of organoids derived from PC patients or pancreatic and lung cancer genetically engineered mouse models. a PC patient and b KPC mouse organoid staining with ch5E6 indicating MUC16 expression (green) and specific binding of mAb compared to no binding with isotype control mAb huIgG1. The binding of ch5E6 to the ductal cells (yellow) was illustrated by colocalization with CK-19 (red) staining in human and mouse pancreatic tumor organoids. Scale bar 10 µm.The representative images for ch5E6 treated. c Human PC patient. d KPC and (e) KPA mouse-derived organoids obtained by real-time kinetics using Incucyte live cell imaging system. The data were quantitated for organoid counts using essence incucyte software, plotted as a graph of change in organoid counts or area over time (3–5 days) for both ch5E6 and huIgG1 treatments, and is shown in parallel. Error bars indicate SEM. *P < 0.05; **P < 0.01.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Derivative Assay, Staining, Expressing, Binding Assay, Control, Live Cell Imaging, Software

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 6 Inhibition of EMT by ch5E6 is validated in PC and NSCLC cell line-derived xenografts. a Immunofluorescence analysis showing a decrease in pFAK(Y397) and N-cadherin expression in xenograft tumors of SW1990 cells treated with ch5E6 compared to isotype control mAb huIgG1 group (n = 6–8 fields/tissue: three animals). The data was plotted for changes in fluorescence intensity using GraphPad Prism 9 and is shown in parallel. Nuclei were stained with DAPI. b Immunofluorescence analysis of ch5E6 treated SW1573 cell line-derived xenografts showing a reduction in pFAK(Y397) and N-cadherin levels compared to isotype control mAb huIgG1 group (n = 6–8 fields/tissue: 3 animals). Scale bar, 10 µm; magnified images, 2 µm. No significant changes in the intensity of MUC16 were seen in the ch5E6 treated versus isotype control tumors derived from both cancers. c, d Immunoblot analysis of ch5E6 treated PDAC and NSCLC tumor lysates showing a substantial decrease in phosphorylated levels of FAK(Y397), p70S6K(T389) and N-cadherin as compared to huIgG1 treatment. Error bars indicate SEM. Scale bar, 400 μm; magnified images, 100 μm; *P < 0.05; **P < 0.01.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Inhibition, Derivative Assay, Expressing, Control, Staining, Western Blot

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 7 MUC16 and N-cadherin are clinically correlated in patient tumors. a, b Representative images and quantitative illustration of immunohistochemical analyses demonstrate a strong positive correlation between MUC16 and N-cadherin (R = 0.84) in both primary PC tumors (n = 10) and liver metastasis (R = 0.99) samples (n = 8). Scale bar, 400 µm; magnified images, 100 µm. *P < 0.05; **P < 0.01. c Representative images of immunofluorescence analysis showing coexpression of MUC16 (green) and N-cadherin (red) in primary PDAC tumors compared to no MUC16 and N-cadherin in normal pancreatic sections. Scale bar, 20 µm; magnified images, 5 µm. d Schematic diagram representing ch5E6 induced downregulation of MUC16 mediated EMT resulting in its anti-tumor potential in PC and NSCLC. Overall, anti- MUC16 chimeric mAb5E6 (ch5E6) binds to the cell surface-tethered domain of MUC16, interferes with oncogenic pFAK/p70S6K/N-cadherin signaling associated with MUC16-mediated EMT, and reduces tumor burden in both PC and NSCLC models.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Immunohistochemical staining

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: MiR-106a was up-regulated in serum of sepsis patients and CLP-induced mice models. The relative miR-106a level was tested by qRT-PCR in the serum from sepsis and healthy patients, CLP-induced sepsis mouse models and TCMK-1 cells treated with LPS. *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Quantitative RT-PCR

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: Knockdown of miR-106a reduced LPS-induced apoptosis in TCMK-1 cells. miR-106a inhibitor, NC inhibitor and LPS were transfected into TCMK-1 cells. (A) Relative miR-106a level, (B) cell viability, (C) relative caspase-3 activity and (D) the protein level of Bax, cleaved caspase-3 and Bcl-2 were detected by qRT-PCR, CCK-8 and western blotting. ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Transfection, Activity Assay, Quantitative RT-PCR, CCK-8 Assay, Western Blot

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: Knockdown of miR-106a reduced LPS-induced inflammatory factor levels in TCMK-1 cells. miR-106a inhibitor, NC inhibitor and LPS were transfected into TCMK-1 cells. (A) The relative mRNA level and (B) content of TNF-α, IL-1β, IL-6 were detected by qRT-PCR and ELISA. * p<0.05; ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: THBS2 was a target of miR-106a. MiR-106a inhibitor, NC inhibitor, and miR-106a mimics, NC mimics, THBS2-WT and THBS2-MUT were transfected into HEK293 and TCMK-1 cells. (A) The binding site was predicted by Targetscan website. (B) Relative miR-106a level was tested by qRT-PCR. (C) The target relationship between miR-106a and THBS2 was confirmed by dual luciferase reporter assay. (D) THBS2 expression was detected by western blotting. ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Transfection, Binding Assay, Quantitative RT-PCR, Luciferase, Reporter Assay, Expressing, Western Blot

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: MiR-106a increased LPS-induced inflammation and apoptosis of TCMK-1 cells via regulating THBS2 expression. MiR-106a inhibitor, NC inhibitor, and miR-106a mimics, NC mimics, siTHBS2 and siNC were transfected into LPS-treated TCMK-1 cells. (A) Relative THBS2 mRNA level, (B) cell viability, (C) relative caspase-3 activity, (D-E) the relative mRNA level and content of TNF-α, IL-1β, IL-6, as well as (F) the protein level of Bax, cleaved caspase-3 and Bcl-2 were detected by qRT-PCR, CCK-8, ELISA and western blotting. * p<0.05; ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Expressing, Transfection, Activity Assay, Quantitative RT-PCR, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot