cell cultures human lung non small cell carcinoma cell line a549  (ATCC)

Journal: Iraqi Journal of Pharmaceutical Sciences( P-ISSN 1683 - 3597 E-ISSN 2521 - 3512)

Article Title: Synergistic Effects of 2-Deoxy-D-Glucose and Cinnamic Acid with Erlotinib on NSCLC Cell Line

doi: 10.31351/vol32isssuppl.pp136-144

Figure Lengend Snippet: Figure 1. The efficacy of 2DG, CINN and ERL on the NSCLC cells viability. A549 cells treated with a range of concentrations of 2DG (A), CINN (B) and ERL. (C) were counted at 48 h using MTT assay. (D) The data are represented as means of triplicate samples for each group. 2DG, 2-Deoxy D glucose; CINN, cinnamic acid; ERL, erlotinib. P<0.0001 (****), P<0.001 (***), P<0.01 (**), P< 0.05 (*). The combination of erlotinib with cinnamic acid or 2DG synergistically repressed the survival of cultured NSCLC cells Erlotinib is well known for having an anticancer effect on a variety of malignancies, thus it is desirable to increase its therapeutic potential at lower doses. In this study, combining non-toxic concentrations (½ IC50 and IC50) of either 2DG or CINN with 15 µM erlotinib in A549 cells for 48 h was performed to assess their synergy. In A549 cells, the combination of 30 µM CINN with 15 µM erlotinib caused 67% decrease in cell viability of A549 cells at 48 h, compared to erlotinib alone(p<0.001). At the same time point, the combination of 2DG (50 mM) with erlotinib exerted a decrease of only 58% compared to erlotinib only treatment (p<0.001).

Article Snippet: Cell cultures Human lung non-small cell carcinoma cell line A549 was obtained from the American Type Culture Collection (ATCC).

Techniques: MTT Assay, Cell Culture

Journal: Journal of Thoracic Disease

Article Title: The impacts of ubiquilin 1 (UBQLN1) knockdown on cells viability, proliferation, and apoptosis are mediated by p53 in A549 lung cancer cells

doi: 10.21037/jtd-20-1362

Figure Lengend Snippet: Effects of UBQLN1 knockdown (KD) on cell viability, proliferation, and apoptosis in A549 cells. (A) Cell viability determined by MTT assay; (B) cell proliferation determined by BrdU assay; (C) cell apoptosis determined by TUNEL assay. Left: representative images showing apoptotic cells; right: quantification of cell apoptosis; (D) representative Western blot images showing apoptosis markers; (E) quantification of cleaved caspase-3 in (D); (F) quantification of protein levels of Bcl-2 and Bax in (D); (G) quantification of Bcl-2/Bax in (D). n=4. **, P<0.01 and ***, P<0.001 vs. siRNA control group. UBQLN1, ubiquilin 1; KD, knockdown.

Article Snippet: Cell culture Human non-small cell lung carcinoma cell line A549 (ATCC ® CCL-185 TM ) and H358 (ATCC ® CRL-5807 TM ), and human normal lung epithelial BEAS-2B cells (ATCC ® CRL-9609 TM ) were purchased from American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, MTT Assay, BrdU Staining, TUNEL Assay, Western Blot, Control

Journal: Journal of Thoracic Disease

Article Title: The impacts of ubiquilin 1 (UBQLN1) knockdown on cells viability, proliferation, and apoptosis are mediated by p53 in A549 lung cancer cells

doi: 10.21037/jtd-20-1362

Figure Lengend Snippet: Effects of p53 overexpression on cell viability, proliferation, and apoptosis in UBQLN1-KD A549 cells. (A) Representative Western blot images showing p53 protein levels; (B) cell viability determined by MTT assay; (C) cell proliferation determined by BrdU assay; (D) cell apoptosis determined by TUNEL assay. Left: representative images showing apoptotic cells; right: quantification of cell apoptosis; (E) representative Western blot images showing apoptosis markers; (F) quantification of cleaved caspase-3 in (E); (G) quantification of Bcl-2/Bax in (E). n=4. ***, P<0.001 vs. siRNA control group; ##, P<0.01 and ###, P<0.001 vs. UBQLN1 KD group. OE, overexpression; UBQLN1, ubiquilin 1; KD, knockdown.

Article Snippet: Cell culture Human non-small cell lung carcinoma cell line A549 (ATCC ® CCL-185 TM ) and H358 (ATCC ® CRL-5807 TM ), and human normal lung epithelial BEAS-2B cells (ATCC ® CRL-9609 TM ) were purchased from American Type Culture Collection (Rockville, MD, USA).

Techniques: Over Expression, Western Blot, MTT Assay, BrdU Staining, TUNEL Assay, Control, Knockdown

Journal: Journal of Thoracic Disease

Article Title: The impacts of ubiquilin 1 (UBQLN1) knockdown on cells viability, proliferation, and apoptosis are mediated by p53 in A549 lung cancer cells

doi: 10.21037/jtd-20-1362

Figure Lengend Snippet: Effects of UBQLN1 knockdown (KD) on the activities of proteasome and autophagy in A549 cells. (A) Effects of UBQLN1 KD on proteasome activity; (B) representative Western blot images showing the effects of UBQLN1 KD on autophagic markers; (C) quantification of LC3-II/LC3-I in (B); (D) representative Western blot images showing the effects of autophagy inhibitor BFA on LC3-II expression; (E) quantification of autophagic flux based on (D); (F) representative Western blot images showing the effects of autophagy inhibitor BFA on p53 protein levels; (G) quantification of p53 protein levels in (F). n=4. **, P<0.01 and ***, P<0.001 vs. siRNA control group; ##, P<0.01 vs. UBQLN1 KD group. UBQLN1, ubiquilin 1; BFA, brefeldin A.

Article Snippet: Cell culture Human non-small cell lung carcinoma cell line A549 (ATCC ® CCL-185 TM ) and H358 (ATCC ® CRL-5807 TM ), and human normal lung epithelial BEAS-2B cells (ATCC ® CRL-9609 TM ) were purchased from American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, Activity Assay, Western Blot, Expressing, Control

Journal: Journal of Thoracic Disease

Article Title: The impacts of ubiquilin 1 (UBQLN1) knockdown on cells viability, proliferation, and apoptosis are mediated by p53 in A549 lung cancer cells

doi: 10.21037/jtd-20-1362

Figure Lengend Snippet: Effects of UBQLN1 knockdown (KD) on ER stress, ROS formation, and the mTOR signaling in A549 cells. (A) Representative Western blot images showing proteins markers of ER stress; (B) representative images showing real time formation of intracellular ROS (magnification ×400); (C) quantification of ROS formation in (B). (D) representative Western blot images showing phosphorylation of mTOR and its downstream S6K; (E) quantification of phosphorylation levels of mTOR and S6K in (C); (F) schematic diagram of UBQLN1 on cell viability, proliferation, and apoptosis in A549 cancer cells. UBQLN1 impairs mTORC1 activity leading to the activation of autophagy and subsequent degradation of p53. Loss of p53 will affect cell viability, proliferation, and apoptosis. n=4. **, P<0.01 and ***, P<0.001 vs. siRNA control group; #, P>0.05 vs. siRNA control group. UBQLN1, ubiquilin 1.

Article Snippet: Cell culture Human non-small cell lung carcinoma cell line A549 (ATCC ® CCL-185 TM ) and H358 (ATCC ® CRL-5807 TM ), and human normal lung epithelial BEAS-2B cells (ATCC ® CRL-9609 TM ) were purchased from American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, Western Blot, Phospho-proteomics, Activity Assay, Activation Assay, Control