Gene expression profiling of lung cancer cells irradiated by carbon ion and X-rays
You, An
Background
Lung cancer is the leading cause of cancer-related death in men and the third in women in Germany. Radiation therapy plays an important role in the multimodal treatment of lung cancer. Due to the excellent dose distribution and the higher relative biological effectiveness (RBE) in tumor, heavy ion therapy with carbon shows promising clinical results in different types of cancer. However, the genetic differences of radiation induced reactions in cancer between heavy ion beams and conventional photon beams are not fully understood. In the present study, we compared the gene expression profiles of A549 cells after heavy ion radiation or X-ray radiation using a DNA microarray chip containing 11,800 human genes and identified differentially expressed genes. A set of selected differentially expressed genes was validated with quantitative real-time polymerase chain reaction (qRT-PCR).
Materials/Methods
The lung carcinoma cell line A549 was irradiated with carbon ion beams (9,8 MeV/nucleon) and X-ray (250 kV) using different doses. The biologically equivalent doses for each radiation quality were determined by clonogenic survival assay. The transcriptional profiling was determined with a high density cDNA microarray containing 11.800 genes, and genetic network and gene ontology analysis was performed. The expression changes of selected genes were validated by qRT-PCR.
Results
Microarray analysis revealed a significant alteration in the expression of 49 genes (at least 2-fold) after carbon ion irradiation and not altered by X-rays, as compared with unirradiated control cells. Of these 49 differentially expressed genes identified, 29 and 20 genes were up- and down-regulated, respectively.
Moreover, the results of microarray analysis showed that the expression levels of 326 genes were altered significantly by carbon ion irradiation with biological equivalent dose to X-rays. Among these genes identified, 169 and 157 genes were more up-and down-regulated in carbon ion irradiation, as compared to X-rays.
The genetic network and functional classification of the 49 differentially expressed genes between carbon ions irradiation and control unirradiated cells revealed four merged networks which were associated with the regulation of cell cycle, cancer and cell death signaling and cell signaling.
The functional analysis of the up-regulated genes between carbon ion and X-ray determined three important functional networks involved in cellular growth and proliferation, cell cycle regulation, and oxidation reduction. Among the down-regulated genes, the functional analysis identified three important molecular functional networks associated with cellular function and maintenance of cancer, regulation of cell cycle in the DNA repair, and post translation modification. A set of 8 selected differentially expressed genes involved in cell cycle, DNA damage and transcription was analysed by qRT-PCR and confirmed the microarray data.
Conclusions
These results showed that these two types of radiations, although in the same biological relative doses, could induce significant gene expression in different levels in A549 cells. The functional classification of these differentially expressed genes revealed that carbon ions and X-ray irradiations have different effects on different signaling pathways through gene expression. The identification of differentially expressed gene in this study might add to the understanding of the complicated molecular responses to carbon ion irradiation and provided valuable resource for both experimental and clinical application of heavy ion beam for treatment of lung cancer.
Philipps-Universität Marburg
Medical sciences Medicine
opus:4581
urn:nbn:de:hebis:04-z2012-09983
https://doi.org/10.17192/z2012.0998
doctoralThesis
2012-11-07
Das Lungenkarzinom ist die häufigste tödliche Krebserkrankung des Mannes und die dritthäufigste tödliche Krebserkrankung der Frau in Deutschland. Die Strahlentherapie spielt eine wichtige Rolle in der multimodalen Behandlung vom Lungenkarzinom. Aufgrund der hervorragenden Dosisverteilung und der höheren relativen biologischen Wirksamkeit (RBW) im Tumor zeigt die Schwerionentherapie mit Kohlenstoff vielversprechende klinische Ergebnisse bei unterschiedlichen Karzinomen. Die genetischen Unterschiede der Strahlenreaktionen im Krebsgewebe nach intensiver Ionenbestrahlung und konventioneller Photonenbestrahlung sind aber bis heute nicht vollständig geklärt. In der vorliegenden Arbeit wurden deshalb die Expressionsprofilen humaner A549 Lungenkarzinomzellen nach Bestrahlung mit Kohlenstoffionen und Röntgenstrahlen mittels eines cDNA Microarrays mit 11.800 menschlichen Genen verglichen und differentiell exprimierten Gene identifiziert. Mit quantitativer Real-Time PCR (qRT-PCR) wurden die Veränderungen der ausgewählten differentiell exprimierten Kandidatengene analysiert.
Die A549 Lungenkarzinomzellen wurden mit Kohlenstoffionen (9,8 MeV/nucleon) und Röntgen (250 kV) bestrahlt. Die biologischen Äquivalentdosen der Kohlenstoffionen und Röntgenstrahlen wurden mit dem klonogenen Überleben-Assay bestimmt.
Im Vergleich zur unbestrahlten Kontrolle zeigte die Mikroarray-Analyse signifikante Veränderungen der Expression von 49 Genen (mindestens 2-fach) nach Bestrahlung mit Kohlenstoff. Davon waren 29 Gene und 20 Gene hoch- und runterreguliert. Anhand der Analyse der Expressionsprofile konnten 326 differentiell exprimierten Gene zwischen Bestrahlung mit Kohlenstoffionen und Röntgenstahlen mit den biologischen Äquivalentdosen identifiziert werden. Im Vergleich zur Röntgenstrahlung waren 169 bzw. 157 Gene nach Bestrahlung mit Kohlenstoffionen signifikanter hoch- und runterreguliert. Die genetische Netzwerk und funktionelle Klassifizierungen der 49 differentiell exprimierten Gene zwischen Kohlenstoffionenstrahlung und unbestrahlter Kontrolle zeigten vier fusionierten Netzwerke, welche in der Regulation des Zellzykluses, des Zelltods, und des Zellsignalwegs beteiligt sind. Weitere funktionelle Analyse der hochregulierten Gene zwischen Kohlenstoffionen und Röntgenstahlen zeigte drei wichtige funktionelle Netzwerke, welche an der Regulation der zellulären Proliferation, des Zellzykluses und der Oxidation beteiligt sind. Die Analyse der runterregulierten Gene zeigte drei wichtige molekulare funktionelle Netzwerke in der Regulation der zellulären Funktion and der Erhaltung des Karzinoms, des Zellzykluses mit der DNA-Reparatur und der posttranskriptionellen Modifizierung. Zur Bestätigung der Mikroarraydaten wurde die Expression der 8 ausgewählten differentiell exprimierten Kandidatengene, welche an der Regulation des Zellzykluses, der DNA-Schädigung und der Transkription beteiligt sind, durch qRT-PCR analysiert.
Die Ergebnisse der vorliegenden Arbeit deuteten darauf hin, dass beide Strahlenqualitäten mit biologischen Äquivalentdosen signifikante unterschiedliche Genexpressionen induzieren und dadurch die unterschiedlichen Wirkungen auf der Regulation der Signaltransduktionswege beeinflussen konnten. Die differentiell expremierten Gene sind an der Regulation der Zellzyklen, DNA-Reparatur und der Oxidierung beteiligt. Die Identifizierung der differentiell exprimierten Gene in der vorliegenden Arbeit kann zum Verständnis der komplizierten molekularen Reaktionen auf Bestrahlung mit Kohlenstoffionen hinzufügen und wertvolle Ressource sowohl für experimentelle als auch für klinische Anwendung der Schwerionentherapie von Lungenkarzinom zur Verfügung stehen.
2012-11-16
monograph
Medical sciences Medicine
Medizin
opus:4581
2012
English
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application/pdf
Medizin
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Universitätsbibliothek Marburg
Expressionsprofilen
urn:nbn:de:hebis:04-z2012-09983
https://doi.org/10.17192/z2012.0998
ths
Prof. Dr.
Keusgen
Michael
Keusgen, Michael (Prof. Dr.)
Expressionsprofilen der Lungenkarzinomzellen nach Bestrahlung mit Kohlenstoffionen und Röntgen
Carbon ion irradiation
Gene expression profiling of lung cancer cells irradiated by carbon ion and X-rays
You, An
You
An
Background
Lung cancer is the leading cause of cancer-related death in men and the third in women in Germany. Radiation therapy plays an important role in the multimodal treatment of lung cancer. Due to the excellent dose distribution and the higher relative biological effectiveness (RBE) in tumor, heavy ion therapy with carbon shows promising clinical results in different types of cancer. However, the genetic differences of radiation induced reactions in cancer between heavy ion beams and conventional photon beams are not fully understood. In the present study, we compared the gene expression profiles of A549 cells after heavy ion radiation or X-ray radiation using a DNA microarray chip containing 11,800 human genes and identified differentially expressed genes. A set of selected differentially expressed genes was validated with quantitative real-time polymerase chain reaction (qRT-PCR).
Materials/Methods
The lung carcinoma cell line A549 was irradiated with carbon ion beams (9,8 MeV/nucleon) and X-ray (250 kV) using different doses. The biologically equivalent doses for each radiation quality were determined by clonogenic survival assay. The transcriptional profiling was determined with a high density cDNA microarray containing 11.800 genes, and genetic network and gene ontology analysis was performed. The expression changes of selected genes were validated by qRT-PCR.
Results
Microarray analysis revealed a significant alteration in the expression of 49 genes (at least 2-fold) after carbon ion irradiation and not altered by X-rays, as compared with unirradiated control cells. Of these 49 differentially expressed genes identified, 29 and 20 genes were up- and down-regulated, respectively.
Moreover, the results of microarray analysis showed that the expression levels of 326 genes were altered significantly by carbon ion irradiation with biological equivalent dose to X-rays. Among these genes identified, 169 and 157 genes were more up-and down-regulated in carbon ion irradiation, as compared to X-rays.
The genetic network and functional classification of the 49 differentially expressed genes between carbon ions irradiation and control unirradiated cells revealed four merged networks which were associated with the regulation of cell cycle, cancer and cell death signaling and cell signaling.
The functional analysis of the up-regulated genes between carbon ion and X-ray determined three important functional networks involved in cellular growth and proliferation, cell cycle regulation, and oxidation reduction. Among the down-regulated genes, the functional analysis identified three important molecular functional networks associated with cellular function and maintenance of cancer, regulation of cell cycle in the DNA repair, and post translation modification. A set of 8 selected differentially expressed genes involved in cell cycle, DNA damage and transcription was analysed by qRT-PCR and confirmed the microarray data.
Conclusions
These results showed that these two types of radiations, although in the same biological relative doses, could induce significant gene expression in different levels in A549 cells. The functional classification of these differentially expressed genes revealed that carbon ions and X-ray irradiations have different effects on different signaling pathways through gene expression. The identification of differentially expressed gene in this study might add to the understanding of the complicated molecular responses to carbon ion irradiation and provided valuable resource for both experimental and clinical application of heavy ion beam for treatment of lung cancer.
Gene expression profiling, Lung cancer
Philipps-Universität Marburg
Bestrahlung mit Kohlenstoffionen
Medizin
2012-11-16
Lungenkarzinomzellen
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