Presentation is loading. Please wait.

Presentation is loading. Please wait.

Genetic Determinants of Pemetrexed Responsiveness and Nonresponsiveness in Non- small Cell Lung Cancer Cells  Ming-Fang Wu, MD, PhD, Yi-Min Hsiao, PhD,

Published byJasper Briggs Modified over 5 years ago

Similar presentations

Presentation on theme: "Genetic Determinants of Pemetrexed Responsiveness and Nonresponsiveness in Non- small Cell Lung Cancer Cells  Ming-Fang Wu, MD, PhD, Yi-Min Hsiao, PhD,"— Presentation transcript:

1 Genetic Determinants of Pemetrexed Responsiveness and Nonresponsiveness in Non- small Cell Lung Cancer Cells  Ming-Fang Wu, MD, PhD, Yi-Min Hsiao, PhD, Chuan-Fu Huang, MD, PhD, Yu-Hsin Huang, MS, Wan-Jung Yang, BS, Hsiu-Wen Chan, MS, Jinghua Tsai Chang, PhD, Jiunn-Liang Ko, PhD  Journal of Thoracic Oncology  Volume 5, Issue 8, Pages (August 2010) DOI: /JTO.0b013e3181e0b954 Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

2 FIGURE 1 Cytotoxic effect of pemetrexed in H460, H1355, H1299, and A549 lung cancer cells and normal lung fibroblast cells MRC-5. Cells (5 × 103 cells/well) were treated with increasing doses of pemetrexed (0, 0.08, 0.4, and 2.0 μM) for 72 hours. Cell viability was measured by MTT assay and the results are presented as the calculated cell growth inhibitory ratio. Experiments were repeated three times. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

3 FIGURE 2 RT-PCR analysis of TS, DHFR, GARFT, RFC, and FPGS mRNA in A549 and H1355 cells. A549 and H1355 cells (8 × 105 cells/60 mm dish) were subjected to varying concentrations of pemetrexed (0, 0.08, 0.4, and 2.0 μM) for 48 hours, and mRNA of TS, DHFR, GARFT, RFC, and β-actin was determined. RT-PCR, reverse transcriptase polymerase chain reaction; TS, thymidylate synthase; DHFR, dihydrofolate reductase; RFC, reduced folate carrier; FPGS, folylpolyglutamate synthase. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

4 FIGURE 3 Cell cycle distribution by flow cytometry in A549 and H1355 cells. A549 and H1355 cells (8 × 105 cells/60 mm dish) were subjected to varying concentrations of pemetrexed (0, 0.08, 0.4, and 2.0 μM) for 48 hours. Cells were resuspended in dulbecco modified Eagle medium (DMEM) at 1 × 106 cells/ml. Cells were detected by flow cytometer and acquired by CellQuest. Data is shown as the percentage of nuclei in the sample and is representative of one of the three experiments performed. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

5 FIGURE 4 Western blot analysis of p16, p21, p27, P53, cyclin, and CDK in A549 and H1355 cells. A549 and H1355 cells (5 × 105 cells/60-mm dish) were subjected to varying concentrations of pemetrexed (0, 0.08, 0.4, and 2.0 μM) for 48 hours. Total amounts of protein expression were determined by immunoblot experiments with the corresponding antibodies. Expression of β-actin on the same immunoblot was used as a loading control. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

6 FIGURE 5 Genetic analysis by superarray. The oligo GEArray system was used to determine the regulation of 480 genes (for a gene table, see Briefly, a total of 3 μg were reverse transcribed into Biotin-16-dUTP-labled cDNA probes with the use of turelabling-AMP 2.0 method, according to the manufacturer's protocol. Cell cultures were incubated in 10 μM pemetrexed-containing or pemetrexed-free medium for 48 hours. A, A549. B, Pemetrexed-treated A549. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

7 FIGURE 6 RT-PCR analysis of Lcn-2 and nm23–H1 mRNA in A549 and H1355 cells after pemetrexed treatment. A549 and H1355 cells (8 × 105) were subjected to varying concentrations of pemetrexed (0, 0.08, 0.4, and 2.0 μM) for 48 hours, and mRNA of Lcn-2, nm23–H1, and β-actin was determined. RT-PCR, reverse transcriptase polymerase chain reaction. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

8 FIGURE 7 Western blot comparisons of Lcn-2, p53, and β-actin expressions in A549 and H1355 cells. A549 and H1355 (5 × 105 cells/60 mm dish) were treated with 0.08, 0.4, and 2 μM pemetrexed for 48 hours, and the total amount of protein expression was determined by immunoblot experiments with the appropriate antibodies. Expression analysis of β-actin served as a loading control. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

9 FIGURE 8 Cytotoxic effects of pemetrexed in A549 Shluc, A549 Shnm23–H1, and A549 ShLcn2 cells. A549 Shluc, A549 Shnm23–H1, and A549 ShLCN2 (5 × 103 cells/well) cells were treated with varying concentrations of pemetrexed (0.08, 0.4, 2, and 10 μM) and MTT assayed after 72 hours of treatment. Cell viability was measured and the results are presented as the calculated cell growth inhibitory ratio. Experiments were repeated three times. Journal of Thoracic Oncology 2010 5, DOI: ( /JTO.0b013e3181e0b954) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

Download ppt "Genetic Determinants of Pemetrexed Responsiveness and Nonresponsiveness in Non- small Cell Lung Cancer Cells  Ming-Fang Wu, MD, PhD, Yi-Min Hsiao, PhD,"

Similar presentations

Supplementary Materials and Methods Real-time RT-PCR Genomic DNA was isolated from washed cell pellets of eight biliary tract cancer cell lines (SNU-245,

Supplementary Materials and Methods Real-time RT-PCR Genomic DNA was isolated from washed cell pellets of eight biliary tract cancer cell lines (SNU-245,
Nicotinamide Phosphoribosyltransferase: A Potent Therapeutic Target in Non-small Cell Lung Cancer with Epidermal Growth Factor Receptor-Gene Mutation 

Nicotinamide Phosphoribosyltransferase: A Potent Therapeutic Target in Non-small Cell Lung Cancer with Epidermal Growth Factor Receptor-Gene Mutation 
IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,

IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,
Raloxifene⁎ increases proliferation of human endothelial cells in association with increased gene expression of cyclins A and B1  Pilar J. Oviedo, B.Sc.,

Raloxifene⁎ increases proliferation of human endothelial cells in association with increased gene expression of cyclins A and B1  Pilar J. Oviedo, B.Sc.,
Aldehyde Dehydrogenase 1A1 Possesses Stem-Like Properties and Predicts Lung Cancer Patient Outcome  Xiao Li, MD, Liyan Wan, MD, Jian Geng, MD, Chin-Lee.

Aldehyde Dehydrogenase 1A1 Possesses Stem-Like Properties and Predicts Lung Cancer Patient Outcome  Xiao Li, MD, Liyan Wan, MD, Jian Geng, MD, Chin-Lee.
Overexpression of CRM1: A Characteristic Feature in a Transformed Phenotype of Lung Carcinogenesis and a Molecular Target for Lung Cancer Adjuvant Therapy 

Overexpression of CRM1: A Characteristic Feature in a Transformed Phenotype of Lung Carcinogenesis and a Molecular Target for Lung Cancer Adjuvant Therapy 
Tumor Cell Repopulation between Cycles of Chemotherapy is Inhibited by Regulatory T- Cell Depletion in a Murine Mesothelioma Model  Licun Wu, MD, Zhihong.

Tumor Cell Repopulation between Cycles of Chemotherapy is Inhibited by Regulatory T- Cell Depletion in a Murine Mesothelioma Model  Licun Wu, MD, Zhihong.
The Autophagy Inhibitor Chloroquine Overcomes the Innate Resistance of Wild-Type EGFR Non-Small-Cell Lung Cancer Cells to Erlotinib  Yiyu Zou, PhD, Yi-He.

The Autophagy Inhibitor Chloroquine Overcomes the Innate Resistance of Wild-Type EGFR Non-Small-Cell Lung Cancer Cells to Erlotinib  Yiyu Zou, PhD, Yi-He.
EZH2 Promotes E2F-Driven SCLC Tumorigenesis through Modulation of Apoptosis and Cell-Cycle Regulation  Roland Hubaux, PhD, Kelsie L. Thu, BSc, Bradley.

EZH2 Promotes E2F-Driven SCLC Tumorigenesis through Modulation of Apoptosis and Cell-Cycle Regulation  Roland Hubaux, PhD, Kelsie L. Thu, BSc, Bradley.
Antitumor Efficacy of the Anti-Interleukin-6 (IL-6) Antibody Siltuximab in Mouse Xenograft Models of Lung Cancer  Lanxi Song, MS, Matthew A. Smith, PhD,

Antitumor Efficacy of the Anti-Interleukin-6 (IL-6) Antibody Siltuximab in Mouse Xenograft Models of Lung Cancer  Lanxi Song, MS, Matthew A. Smith, PhD,
Group IIa secretory phospholipase expression correlates with group IIa secretory phospholipase inhibition–mediated cell death in K-ras mutant lung cancer.

Group IIa secretory phospholipase expression correlates with group IIa secretory phospholipase inhibition–mediated cell death in K-ras mutant lung cancer.
PI3K as a Potential Therapeutic Target in Thymic Epithelial Tumors

PI3K as a Potential Therapeutic Target in Thymic Epithelial Tumors
Upregulation of PD-L1 by EGFR Activation Mediates the Immune Escape in EGFR- Driven NSCLC: Implication for Optional Immune Targeted Therapy for NSCLC Patients.

Upregulation of PD-L1 by EGFR Activation Mediates the Immune Escape in EGFR- Driven NSCLC: Implication for Optional Immune Targeted Therapy for NSCLC Patients.
Berberine inhibits the proliferation of human uterine leiomyoma cells

Berberine inhibits the proliferation of human uterine leiomyoma cells
WT1 Promotes Invasion of NSCLC via Suppression of CDH1

WT1 Promotes Invasion of NSCLC via Suppression of CDH1
Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.

Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.
MET Tyrosine Kinase Inhibitor Crizotinib (PF-02341066) Shows Differential Antitumor Effects in Non-small Cell Lung Cancer According to MET Alterations 

MET Tyrosine Kinase Inhibitor Crizotinib (PF ) Shows Differential Antitumor Effects in Non-small Cell Lung Cancer According to MET Alterations 
Droxinostat, a Histone Deacetylase Inhibitor, Induces Apoptosis in Hepatocellular Carcinoma Cell Lines via Activation of the Mitochondrial Pathway and.

Droxinostat, a Histone Deacetylase Inhibitor, Induces Apoptosis in Hepatocellular Carcinoma Cell Lines via Activation of the Mitochondrial Pathway and.
E1B-55 kDa-Defective Adenoviruses Activate p53 in Mesothelioma and Enhance Cytotoxicity of Anticancer Agents  Makako Yamanaka, MD, Yuji Tada, MD, PhD,

E1B-55 kDa-Defective Adenoviruses Activate p53 in Mesothelioma and Enhance Cytotoxicity of Anticancer Agents  Makako Yamanaka, MD, Yuji Tada, MD, PhD,
Constitutive and tumor necrosis factor-α-induced activation of nuclear factor-κB in adenomyosis and its inhibition by andrographolide  Bin Li, M.S., Ming.

Constitutive and tumor necrosis factor-α-induced activation of nuclear factor-κB in adenomyosis and its inhibition by andrographolide  Bin Li, M.S., Ming.

Similar presentations

Ads by Google