human lung cancer cell line pc9  (Millipore)

Journal: Translational Oncology

Article Title: NTRK2 promotes malignant progression and paclitaxel resistance of lung adenocarcinoma through targeting MYC/ABCF1 axis

doi: 10.1016/j.tranon.2025.102655

Figure Lengend Snippet: NTRK2 promotes malignant behavior of LUAD cells in vitro . (A) The expression of NTRK2 was verified by qRT-PCR at the transcriptional level in A549, A549/Taxol and PC9 cells. (B) Western blotting analysis was performed to validate the expression of NTRK2 protein in the above cells. (C) MTT showing the effect of NTRK2 knockdown on the proliferation of A549, A549/Taxol and PC9 cells. Data are shown as mean ± SD. Statistical analysis was performed by two-way ANOVA. (D) The effect of NTRK2 knockdown on colony formation ability of the above cells. The right histograms showed the number of colonies for each cell line as indicated. (E) Relative mRNA expression level of NTRK2 was determined by qRT-PCR in A549 and PC9 cells transfected with an empty vector (Vector) or an NTRK2-overexpressing plasmid (oe-NTRK2). (F) Protein expression level of NTRK2 was analyzed by Western blotting in the same set of transfected cells. (G) Cell proliferation was assessed by MTT assay in A549 and PC9 cells following NTRK2 overexpression. Data are presented as the mean ± SD. Statistical analysis was performed by two-way ANOVA. (H) The promoting effect of NTRK2 overexpression on colony formation in A549 and PC9 cells. The effect of NTRK2 knockdown on cell cycle distributions (I) and apoptosis (J) was examined by the flow cytometry. The histograms of cell cycle distributions and apoptosis are presented on the right side of their respective representative graphs. Data are presented as the mean ± SD. β-actin was used as an internal control for qRT-PCR. GAPDH was adopted as the internal control for Western blotting. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human LUAD cell lines PC9, A549 and its taxol-resistant strain A549/Taxol were purchased from Procell Life Science & Technology Co.,Ltd (Wuhan, China).

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Transfection, Plasmid Preparation, MTT Assay, Over Expression, Flow Cytometry, Control

Journal: Translational Oncology

Article Title: NTRK2 promotes malignant progression and paclitaxel resistance of lung adenocarcinoma through targeting MYC/ABCF1 axis

doi: 10.1016/j.tranon.2025.102655

Figure Lengend Snippet: NTRK2 reduces the response of LUAD cells to paclitaxel in vitro. NTRK2-knockdown A549 and A549/Taxol cells (A), NTRK2-overexpressing A549 and PC9 cells (B) as well as control cells were subjected to dose-dependent paclitaxel treatment for 48 h. The cell viability was assessed via MTT assay, and the Reed-Muench method was adopted to calculate IC 50 values. (C) The cells with NTRK2 knockdown or overexpression mentioned above and their control cells were subjected to treatment with paclitaxel at indicated concentrations for a duration of 4 days or treatment with DMSO. MTT assays were conducted to evaluate the time-dependent proliferation of these cells. Data are shown as mean ± SD. Statistical analysis was performed by two-way ANOVA. NTRK2-knockdown A549/Taxol (D) and NTRK2-overexpressing A549 cells (E) were treated with paclitaxel at indicated concentrations to evaluate the effect of NTRK2 expression on the inhibitory effect of paclitaxel on clone formation. A549/Taxol cells with NTRK2 knockdown were treated with paclitaxel or DMSO for 24 h, and cell cycle distributions (F) and apoptosis (G) were analyzed by the flow cytometry. ** P < 0.01; *** P < 0.001.

Article Snippet: Human LUAD cell lines PC9, A549 and its taxol-resistant strain A549/Taxol were purchased from Procell Life Science & Technology Co.,Ltd (Wuhan, China).

Techniques: In Vitro, Knockdown, Control, MTT Assay, Endpoint Dilution Assay, Over Expression, Expressing, Flow Cytometry

Journal: Translational Oncology

Article Title: NTRK2 promotes malignant progression and paclitaxel resistance of lung adenocarcinoma through targeting MYC/ABCF1 axis

doi: 10.1016/j.tranon.2025.102655

Figure Lengend Snippet: ABCF1 is a key target of NTRK2 in mediating resistance to paclitaxel in LUAD cells. The mRNA (A) and protein (B) expression levels of NTRK2 and ABCF1 in A549 and A549/Taxol cells were measured by qRT-PCR and Western blotting analysis, respectively. (C) Representative immunohistochemical staining of NTRK2 and ABCF1 in LUAD tissues (Case 1), paclitaxel-resistant LUAD tissues (Case 2) and their matched noncancerous lung tissues (MN). Scale bars: 50 μm. qRT-PCR (D) and Western blotting (E) analysis results showing the effect of NTRK2 knockdown on ABCF1 expression in A549 and A549/Taxol cell lines. qRT-PCR (F) and Western blotting (G) analysis results showing the effect of ectopic expression of NTRK2 on ABCF1 expression in A549 and PC9 cells. (H) ABCF1-knockdown A549, A549/Taxol cells and control cells were subjected to dose-dependent paclitaxel treatment for 48 h. The cell viability was assessed via MTT assay, and the Reed-Muench method was adopted to calculate IC 50 values. (I) Knockdown of ABCF1 in A549 cells cooperated with overexpressing NTRK2 and their control cells were subjected to 0, 5 and 10 nM paclitaxel treatment for 48 h. The relative cell viability was assessed by MTT assay. β-actin served as an internal control for qRT-PCR. GAPDH was the internal reference for Western blotting. Data are expressed as mean ± SD. All above experiments were independently repeated in triplicate. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human LUAD cell lines PC9, A549 and its taxol-resistant strain A549/Taxol were purchased from Procell Life Science & Technology Co.,Ltd (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining, Knockdown, Control, MTT Assay, Endpoint Dilution Assay

Journal: Translational Oncology

Article Title: NTRK2 promotes malignant progression and paclitaxel resistance of lung adenocarcinoma through targeting MYC/ABCF1 axis

doi: 10.1016/j.tranon.2025.102655

Figure Lengend Snippet: NTRK2 increases ABCF1 expression through MYC. (A) Western blotting analysis showing the effect of NTRK2 knockdown on the activity of PI3K/AKT and MAPK/ERK pathways and MYC expression in A549, A549/Taxol and PC9 cells. (B) The effect of NTRK2 overexpression on the activity of PI3K/AKT and MAPK/ERK pathways and MYC expression in A549 and PC9 cells. The effect of MYC inhibitor 10058-F4 on cell proliferation (C) and colony formation (D) of A549 cells cooperated with overexpressing NTRK2 . (E) Analysis the correlation between MYC and ABCF1 in lung cancer tissues (the data from TCGA and GTEx datasets). A549 and PC9 cells were treated with 60 μM 10058-F4 or DMSO for 24 h, and the mRNA (F) and protein (G) expression of MYC and ABCF1 were detected. A549 and PC9 cells were transfected with an empty vector (Vector) or an MYC-overexpressing plasmid (oe-MYC), the mRNA (H) and protein (I) expression levels of MYC and ABCF1 were determined by qRT-PCR and Western blotting assays. (J) The impact of MYC overexpression on the promoter activity of the wild-type (WT) and mutant (Mut) ABCF1 was evaluated using a luciferase reporter system in A549 and PC9 cells. ABCF1-Luc reporter activity was normalized to Renilla activity (Luc/Renilla). The Luc/Renilla activity was represented as means ± SD (n = 3). (K) ChIP-qPCR assay shows MYC binding to three different regions of ABCF1 promoter in A549 and PC9 cells ( n = 3). (L) qRT-PCR assay was performed to detect the mRNA expression of both MYC (left panel) and ABCF1 (right panel) in NTRK2-overexpressed cells after treating with 10058-F4 or DMSO. (M) Western blotting analysis of NTRK2, MYC and ABCF1 in NTRK2-overexpressed A549 and PC9 cells treated with 60 μM 10058-F4. β-actin was the normalized control for qRT-PCR and GAPDH served as the internal reference for Western blotting analysis. Data are shown as the mean ± SD. All above experiments were independently repeated in triplicate. ** P < 0.01; *** P < 0.001.

Article Snippet: Human LUAD cell lines PC9, A549 and its taxol-resistant strain A549/Taxol were purchased from Procell Life Science & Technology Co.,Ltd (Wuhan, China).

Techniques: Expressing, Western Blot, Knockdown, Activity Assay, Over Expression, Transfection, Plasmid Preparation, Quantitative RT-PCR, Mutagenesis, Luciferase, ChIP-qPCR, Binding Assay, Control

Journal: Cancer Research

Article Title: The circRNA cEMSY Induces Immunogenic Cell Death and Boosts Immunotherapy Efficacy in Lung Adenocarcinoma

doi: 10.1158/0008-5472.CAN-24-1484

Figure Lengend Snippet: cEMSY expression level correlates with an enhanced immunotherapy response. A, Illustrative images of 65 lung adenocarcinoma (LUAD) tissues showing cEMSY (magenta), CD8 + T cells (green), CD11c + DCs (white), and DAPI (blue). Samples were categorized based on cEMSY fluorescence into cEMSY high (above the mean intensity) and cEMSY low (below the mean intensity). B, Analysis of the proportion of intratumor CD11c + DCs and CD8 + T cells within DAPI + cells across the two cEMSY stratifications. Data presented as median ± quartiles for groups cEMSY low ( n = 37) and cEMSY high ( n = 28). C, The Kaplan–Meier curve for the disease-free survival (DFS) of patients with LUAD, who were divided into two groups according to the mean expression of cEMSY. D, Heatmap showing Spearman correlations between CD11c + DCs and CD8 + T-cell infiltration and cEMSY signature gene set variation analysis score of indicated gene sets in each TCGA cohort. ACC, adrenocortical carcinoma; BLCA, bladder urothelial Carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; ESCA, esophageal carcinoma; GBM, glioblastoma multiforme; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LGG, lower grade glioma; LIHC, liver hepatocellular carcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, tescticular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma. E, cEMSY signature expression level of the responders (R) and nonresponders (NR) of two representative LUAD ICB cohorts. F, Progression-free survival (PFS) of ICB cohorts based on the level of cEMSY signature. Statistics were applied using Spearman rank correlation, Student t test, or log-rank test.

Article Snippet: Human bronchial epithelial (HBE) cells and human lung adenocarcinoma cell lines (H358, A549, H23, H1975, PC9, A427, and H373), as well as murine lung adenocarcinoma cell lines (LLC1 and LA795), were obtained from the China Center for Type Culture Collection.

Techniques: Expressing, Fluorescence