tnf α neutralizing antibody infliximab  (MedChemExpress)

Journal: Clinical and Translational Medicine

Article Title: miR‐941 in extracellular vesicles confers anlotinib resistance via Keap1/Nrf2 axis and represents a therapeutic target in non‐small cell lung cancer

doi: 10.1002/ctm2.70721

Figure Lengend Snippet: The miR‐941/ Keap1 / Nrf2 axis promotes malignant phenotypes of proliferation and migration. (A, B) miR‐941 upregulates MMP9 and MMP27 at the mRNA (A) and protein (B) levels in A549 cells. (C and D) miR‐941 promotes cell migration in an Nrf2‐dependent manner. (C) Representative wound healing images in A549 cells transfected with miR‐941 mimic or NC, with or without ML385 (5 µM). Scale bar, 200 µm. (D) Quantification of wound closure. (E and F) miR‐941 upregulates TNF‐α mRNA (E) and protein (F) levels. (G) Pro‐proliferative effect of miR‐941 is mediated by secreted TNF‐α. Cell viability of A549 cells treated with conditioned medium from miR‐941 ‐overexpressing cells, with or without Infliximab. (H and I) Pro‐migratory effect requires intact Keap1/Nrf2 axis. (H) The wound healing assay of cells treated with miR‐941/WT medium, miR‐941/KEAP1 mut medium and miR‐941/NRF2 KO medium. Scale bar, 200 µm. (I) Quantification of wound closure. (J and K) TNF‐α upregulation depends on Keap1/Nrf2 axis, measured by RT‐qPCR (J) and ELISA (K) in Keap1‐mutant and Nrf2‐KO cells. (L and M) Transwell invasion assays in A549 cells transfected with miR‐941 mimic or NC. (L) Representative images. Scale bar, 100 µm. (M) Quantification of migrated cells per field. Error bars represent standard deviation ( n = 3). p ‐values indicated were calculated by Student's t ‐test (unpaired). ns: p > 0.05, * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: For in vitro treatments, the following reagents were used: anlotinib (MCE, Cat#HY‐101416), ML385 (MCE, Cat#HY‐100523; specific Nrf2 inhibitor), the TNF‐α neutralizing antibody infliximab (MCE, Cat#HY‐P9970), and the pan‐caspase inhibitor Z‐VAD‐FMK (MCE, Cat#HY‐16658B, 20 μM, 24 h).

Techniques: Migration, Transfection, Wound Healing Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Mutagenesis, Standard Deviation

Journal: Clinical and Translational Medicine

Article Title: miR‐941 in extracellular vesicles confers anlotinib resistance via Keap1/Nrf2 axis and represents a therapeutic target in non‐small cell lung cancer

doi: 10.1002/ctm2.70721

Figure Lengend Snippet: The miR‐941/ Keap1 / Nrf2 signalling signature is active in anlotinib‐resistant patient tumours. (A) RT‐qPCR analysis showing elevated miR‐941 levels in tumour tissues from patients resistant to anlotinib compared to sensitive patients. (B) Representative Western blots and quantitative results showing downregulation of Keap1 and upregulation of Nrf2 and its downstream targets ( Bcl‐2 , Mcl‐1 , Bcl‐xL , TNF‐α , MMP9 , MMP27 ) in resistant tumours. (C–E) RT‐qPCR analysis confirming upregulation of the mRNA levels of (C) anti‐apoptotic genes ( Bcl‐2 , Mcl‐1 , Bcl‐xL ), (D) TNF‐α , and (E) MMPs ( MMP9 , MMP27 ) in resistant versus sensitive patient tumours. (F, G) Kaplan–Meier survival curves showing (F) progression‐free survival (PFS) and (G) overall survival (OS) of anlotinib‐treated NSCLC patients stratified by tumour miR‐941 expression (high vs. low, based on median expression value). Data are presented as mean ± SEM. p ‐values indicated were calculated by Mann–Whitney U tests. ns: p > 0.05, * p < 0.05, ** p < 0.01.

Article Snippet: For in vitro treatments, the following reagents were used: anlotinib (MCE, Cat#HY‐101416), ML385 (MCE, Cat#HY‐100523; specific Nrf2 inhibitor), the TNF‐α neutralizing antibody infliximab (MCE, Cat#HY‐P9970), and the pan‐caspase inhibitor Z‐VAD‐FMK (MCE, Cat#HY‐16658B, 20 μM, 24 h).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, MANN-WHITNEY