human non small cell lung cancer nsclc adenocarcinoma derived cell line a549  (ATCC)

Journal: Oncology Research

Article Title: Cisplatin-induced activation of TGF-β signaling contributes to drug resistance

doi: 10.32604/or.2023.030190

Figure Lengend Snippet: Effect of cisplatin on various types of cancer cell lines. (A) Cisplatin enhanced N-cadherin expression in A549 but not in DLD-1, HCT116, HT29, SW48, and LNCaP cells. Cells were treated with 1 μM cisplatin for the indicated periods (0, 3, 6, 9, and 12 days). The protein expression of N-cadherin and β-actin was detected by western blotting. Images are representative of two independent experiments. (B) Cisplatin decreased E-cadherin mRNA, and increased N-cadherin and Slug mRNAs in A549 cells. A549 cells were treated with 1 μM cisplatin for the indicated periods. The amount of E-cadherin, N-cadherin, and Slug mRNAs were evaluated using qPCR. The presented data are normalized to Day 0. All data are representative of three independent experiments and presented as mean SD ± (n = 3). * p < 0.05 and ** p < 0.01 (two-tailed Student’s test). (C) The TβRI kinase inhibitor, SB431452, partially suppressed cisplatin-induced Slug upregulation in A549 cells. A549 cells were treated with 10 μM cisplatin and/or 1 μM SB431452. Twenty-four hours later, the cells were collected and subjected to qPCR analysis to detect Slug mRNA. The presented data are normalized to control cells. All data are representative of three independent experiments and presented as mean SD ± (n = 3). * p < 0.05 and ** p < 0.01 (two-tailed Student’s test). (D) TGF-β1 induces and decreases N-cadherin and E-cadherin in LoVo and A549 cells. Cells were stimulated with the indicated concentrations of TGF-β1 (0, 0.1, 1, 10 ng/mL). Two days later, the cells were collected and subjected to western blot analysis to detect E-cadherin, N-cadherin, and β-actin.

Article Snippet: Human colorectal adenocarcinoma-derived cell line HCT116, HT29, and DLD-1, human non-small cell lung cancer (NSCLC) adenocarcinoma-derived cell line A549, and human prostate cancer-derived cell line LNCaP were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Expressing, Western Blot, Two Tailed Test, Control

Journal: Oncology Research

Article Title: Cisplatin-induced activation of TGF-β signaling contributes to drug resistance

doi: 10.32604/or.2023.030190

Figure Lengend Snippet: Cisplatin enhances the secretion of TGF-β1. (A) Cisplatin increased the amount of TGF-β1 in the culture media from LoVo and A549 cells. LoVo and A549 cells were treated with 1 μM cisplatin for 72 h. The culture media from cells were collected and subjected to ELISA to measure the amount of TGF-β1. The amounts of TGF-β1 from LoVo and A549 cells without cisplatin were used to calculate relative values. All data are presented as mean SD ± (n = 3). *** p < 0.001 (two-tailed Student’s test). (B) The amount of TGF-β1 in the culture media from cisplatin r /LoVo cells were increased. The culture media from LoVo or cisplatin r /LoVo (#10, #19, and #54) cells were collected and subjected to ELISA to measure the amount of TGF-β1. The amount of TGF-β1 from LoVo cells without cisplatin was used to calculate relative values. All data are presented as mean SD ± (n = 3). ** p < 0.01 and *** p < 0.001 (two-tailed Student’s test). (C) The effect of cisplatin on the expression of TGF-β1 mRNA in LoVo cells. LoVo cells were treated with 1 μM cisplatin. Three days later, the total RNA was extracted and subjected to qPCR analysis to measure the amount of TGF-β1 mRNA. The presented data are normalized to control cells. All data are representative of three independent experiments. (D) The comparison of the expression levels of TGF-β1 mRNA in LoVo and cisplatin r /LoVo cells. Total RNA from exponentially growing LoVo or cisplatin r /LoVo cells (#10, #19, and #54) was extracted and subjected to qPCR analysis to measure the amount of TGF-β1 mRNA. The presented data are normalized to LoVo cells. All data are representative of three independent experiments.

Article Snippet: Human colorectal adenocarcinoma-derived cell line HCT116, HT29, and DLD-1, human non-small cell lung cancer (NSCLC) adenocarcinoma-derived cell line A549, and human prostate cancer-derived cell line LNCaP were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test, Expressing, Control, Comparison

Journal: Cancer Cell International

Article Title: EF24 exerts cytotoxicity against NSCLC via inducing ROS accumulation

doi: 10.1186/s12935-021-02240-z

Figure Lengend Snippet: EF24 inhibits the growth of NSCLC cells in-vitro. A A549, SPC-A1, H460 and H520 cells were treated with the indicated concentrations of EF24 for 24 h and 48 h, and then subjected to MTT assay. The absorbance value was calculated and standardized to the control group. B The cells were treated with 0 μM, 1 μM, 2 μM and 4 μM EF24 respectively for 2 h and subjected to the cell colony formation assay. Surviving fraction is presented as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.01

Article Snippet: Human non-small cell lung cancer (NSCLC) derived cell lines A549, SPC-A1, H460 and H520 were purchased from American Type Culture Collection (Manassas, VA, USA) and China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vitro, MTT Assay, Control, Colony Assay

Journal: Cancer Cell International

Article Title: EF24 exerts cytotoxicity against NSCLC via inducing ROS accumulation

doi: 10.1186/s12935-021-02240-z

Figure Lengend Snippet: EF24 inhibits tumor growth in-vivo. A A brief flowchart of the experiment design in-vivo. A549 xenograft tumor was established and divided into the following four groups and received 17-day treatment: control, EF24 (5 mg/kg/d, 10 mg/kg/d and 20 mg/kg/d). B The weight of tumors. C The curves of tumor growth. D The body weight of mice. E Representative images of xenografts from different groups. F H&E staining tumor tissue sections of different groups. Magnification: 100×. G Anti-Ki-67 staining of tumor tissues in different groups. Magnification: 200×. The results shown are means ± SD; * P < 0.05

Article Snippet: Human non-small cell lung cancer (NSCLC) derived cell lines A549, SPC-A1, H460 and H520 were purchased from American Type Culture Collection (Manassas, VA, USA) and China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vivo, Control, Staining

Journal: Cancer Cell International

Article Title: EF24 exerts cytotoxicity against NSCLC via inducing ROS accumulation

doi: 10.1186/s12935-021-02240-z

Figure Lengend Snippet: Histopathological analyses of major organs. Histopathological analyses of liver, lung, spleen, kidney and heart tissues of A549 xenograft. Scale bars: 50 μm

Article Snippet: Human non-small cell lung cancer (NSCLC) derived cell lines A549, SPC-A1, H460 and H520 were purchased from American Type Culture Collection (Manassas, VA, USA) and China Center for Type Culture Collection (Wuhan, China).

Techniques:

Journal: Cancer Cell International

Article Title: EF24 exerts cytotoxicity against NSCLC via inducing ROS accumulation

doi: 10.1186/s12935-021-02240-z

Figure Lengend Snippet: EF24 induces apoptosis in NSCLC cells. A A549, SPC-A1, H460 and H520 cells were treated with or without indicated concentrations of EF24 respectively for 48 h and subjected to apoptosis assay using Annexin-V & PI staining. B The expression of apoptotic marker Fas in cells were determined. Three independent experiments were performed and the results statistically analyzed as means ± SD, * P < 0.05, ** P < 0.01, *** P < 0.01

Article Snippet: Human non-small cell lung cancer (NSCLC) derived cell lines A549, SPC-A1, H460 and H520 were purchased from American Type Culture Collection (Manassas, VA, USA) and China Center for Type Culture Collection (Wuhan, China).

Techniques: Apoptosis Assay, Staining, Expressing, Marker

Journal: Cancer Cell International

Article Title: EF24 exerts cytotoxicity against NSCLC via inducing ROS accumulation

doi: 10.1186/s12935-021-02240-z

Figure Lengend Snippet: EF24 induces ROS accumulation, mitochondrial morphological changes and autophagy. A – B EF24 induces ROS accumulation. C EF24 induced mitochondrial fragmentation of the mitochondrial network in A549 cells. The arrow indicates fragmented mitochondria. D Western blot analysis demonstrated SQSTM1, LC3B and ACTB expression in EF24-treated A549 cells. E Autophagic vacuoles in A549 cells treated with various concentrations of EF24 were observed by TEM. The arrow indicates autophagic vacuoles. Number of autophagic vacuoles were calculated. F Changes in the localization of LC3B in cells assessed by immunofluorescence under confocal laser microscopy. The results are presented as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.01

Article Snippet: Human non-small cell lung cancer (NSCLC) derived cell lines A549, SPC-A1, H460 and H520 were purchased from American Type Culture Collection (Manassas, VA, USA) and China Center for Type Culture Collection (Wuhan, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Microscopy

Journal: Cancer Cell International

Article Title: EF24 exerts cytotoxicity against NSCLC via inducing ROS accumulation

doi: 10.1186/s12935-021-02240-z

Figure Lengend Snippet: EF24 induces cytotoxicity is restored by ROS scavengers. A549 and H520 cells were treated with 4 μM EF24 for 48 h with or without CAT and NAC employed 2 h before EF24 treatment, after that cell viability using MTT assay ( A ) and apoptotic cells using Annexin-V & PI staining ( B ) were detected. Three independent experiments were performed and the results statistically analyzed as means ± SD, * P < 0.05, ** P < 0.01, *** P < 0.01

Article Snippet: Human non-small cell lung cancer (NSCLC) derived cell lines A549, SPC-A1, H460 and H520 were purchased from American Type Culture Collection (Manassas, VA, USA) and China Center for Type Culture Collection (Wuhan, China).

Techniques: MTT Assay, Staining

Journal: Nature Communications

Article Title: Distinct initiating events underpin the immune and metabolic heterogeneity of KRAS -mutant lung adenocarcinoma

doi: 10.1038/s41467-019-12164-y

Figure Lengend Snippet: PPP blockade abrogates the growth of KEAP1 -mutant LUAD. a Schematic of spontaneous tumor treatment study. Briefly, KK or KP mice were randomized into 20 mg/kg 6-AN or vehicle treated 40 days post Ad5-CMV-Cre intranasal infection. Lungs were harvested for analysis on day 64. b Representative H&E images of KK vehicle or 6-AN-treated lungs. Scale, 1 mm. c Quantification of KK superior lobe lung weight (milligrams, mg) following vehicle ( n = 6) or 6-AN ( n = 8) treatment. Unpaired t test * p = 0.0106. Mean ± SEM. d Ratio of KK hyperplasia relative to large airway size in vehicle ( n = 6) and 6-AN ( n = 8) treated lungs. Unpaired t test * p = 0.0116. Mean ± SEM. e Colony assay 72 h following treatment with 62.5 μM 6-AN vs vehicle in KEAP1 MUT and KEAP1 WT NSCLC cell lines. Scale, 5 mm. f Relative change in tumor size of xenograft models ( n = 3/cell line) of KEAP1 MUT and KEAP1 WT 48 h following treatment with 20 mg/kg 6-AN or vehicle. Change in size relative to vehicle of each cell line. Two-way ANOVA/Sidak’s multiple comparisons test: A549 ** p = 0.0012; H460 *** p = 0.001. Mean ± SD. g Tumor measurements of A549 cell line xenograft following treatment with 20 mg/kg 6-AN ( n = 6) or vehicle ( n = 6) every 10 days. Survival log-rank (Mantel-Cox) test ** p = 0.0023. h Kaplan–Meier survival curve of A549 xenograft NSG mice treated with 20 mg/kg 6-AN ( n = 6) or vehicle ( n = 6) every 10 days. Mantel–Cox test ** p = 0.0017. i Tumor measurements of H358 xenograft following treatment with 20 mg/kg 6-AN ( n = 8) or vehicle ( n = 4) every 10 days. j Kaplan–Meier survival curve of H358 xenograft NSG mice treated with 20 mg/kg 6-AN ( n = 8) or vehicle ( n = 4) every 10 days

Article Snippet: The human lung cancer derived cell lines A549, H460, H441 and H358 were obtained from ATCC and cultured in RPMI-1640 + GlutaMAX medium (Gibco) supplemented with 10 % FBS (Sigma-Aldrich), 100 U/mL penicillin 100 μg/mL streptomycin (Gibco).

Techniques: Mutagenesis, Infection, Colony Assay