2011 4 31 2 Chin J Radiol Med Prot April 2011 Vol. 31 No. 2 167 CNE-2R CNE-2 CNE-2R Human-6v3. 0 2 CNE-2R CNE-2 2 374 Kegg Biocarta Toll IL-1R 13 JUN MYD88 CCL5 CXCL10 STAT1 LY96 FOS CCL3 IL-6 IL-8 IL-1α IL- 1β IRAK2 t = 13. 47 ~ 66. 57 P < 0. 05 Toll IL-1R Radioresistance-related signaling pathways in nasopharyngeal carcinoma cells GUO YaZHU XiaodongQU SongSU FangWANG QiZHANG Wei. Cancer Hospital of Guangxi Medical University Cancer Institute of Guangxi Zhuang Autonomous RegionNanning 530021China Corresponding author ZHU Xiao-dongEmail zhuxiaodong1966@ yahoo. com. cn Abstract Objective To study the difference of gene expression profile between the radioresistant human nasopharyngeal carcinoma cell line CNE-2R and CNE-2and to screen the signaling pathway associated with radioresistance of nasopharyngeal carcinoma. Methods The radioresistant nasopharyngeal carcinoma cell line CNE-2R was constructed from the original cell line CNE-2. CNE-2R and CNE-2 cells were cultured and administered with 60 Co γ-ray irradiation at the dose of 400 cgy for 15 times. Human-6v 3. 0 whole genome expression profile was used to screen the differentially expressed genes. Bioinformatic analysis was used to identify the pathways related to radioresistance. Results The number of the differentially expressed genes that were found in these 2 experiments was 374. The Kegg pathway and Biocarta pathway analysis of the differentially expressed genes showed the biological importance of Toll-like receptor signaling pathway and IL-1R-mediated signal transduction pathway to the radioresistance of the CNE-2R cells and the significant differences of 13 genes in these 2 pathwaysincluding JUNMYD88 CCL5CXCL10STAT1LY96FOSCCL3IL-6IL-8IL-1αIL-1βand IRAK2 t = 13. 47-66. 57 P < 0. 05. Conclusions Toll-like receptor signaling pathway and IL-1R-mediated signal transduction pathway might be related to the occurrence of radioresistance. Key words Nasopharyngeal carcinoma Signaling pathway Radioresistance Gene chip nasopharyngeal carcinoma NPC 1 5 1 30% 20% DOI 10. 3760 / cma. j. issn. 0254-5098. 2011. 02. 013 30860329 0832229 2010 2010105981002M214 851-2009-055 530021 Email zhuxiaodong1966@ yahoo. com. cn 2-5 3-6 Kegg Biocarta
168 2011 4 31 2 Chin J Radiol Med Prot April 2011 Vol. 31 No. 2 5. RNA 80% ~ 90% RNA 1. CNE-2 Cat1791 cdna crna 7. 1. 5 μg 58 CNE-2R 16 ~ 20 h RPMI 1640 Gibco 1 2 Trizol streptabidin Cy3 3 Invitrogen Human 6v3. 0 8. Bead Station Illumina BeadChip Reader BeadStation Illumina BeadStudio Bead Studio 2. 1 10% RPMI 1640 CNE-2R CNE-2 37 5% CO 2 CNE-2R Diffscore 3. - 20 + 2 CNE-2 60 Co γ 400 cgy 30 cm 30 cm 80 cm Biocarta 94. 45 cgy / min enrichment 2 γ 400 cgy 15 CNE-2R LQ SF = e - αd + βd2 30 α β d 4. 2 CNE-2 0. 25% CNE-2R α β MID SF 2 χ 2 SF 2 6 37 3 cubic spline CO 2 10 ~ 14 d Kegg Biocarta χ 2 30 min 13 t 30 min 50 P < 0. 05 3 SF = / 1. CNE-2R CNE-2 PE = / CNE-2 CNE-2R 3 RNA A 260 / A 280 RNA 6. RNA RNA Kegg 9. SPSS 13. 0 CNE-2 CNE-2R χ 2 = 19. 163
2011 4 31 2 Chin J Radiol Med Prot April 2011 Vol. 31 No. 2 169 P < 0. 051 CNE-2R CNE-2 1 1 CNE-2R CNE-2 α Gy - 1 β Gy - 2 α / β SF 2 CNE-2 0. 537 0. 024 22. 375 0. 310 CNE-2R 0. 166 0. 048 3. 458 0. 592 a a CNE-2 χ 2 = 19. 163 P < 0. 05 1 2 2 1 2 2 2 RNA Kegg Biocarta Kegg Toll 2 Biocarta IL-1R 2 2 13 JUN MYD88 CCL5 CXCL10 STAT1 LY96 FOS CCL3 IL-6 IL-8 IL-1α IL-1β IRAK2 t = 13. 47 ~ 66. 57 1 CNE-2R CNE-2 P < 0. 053 2. RNA 2 CNE-2R CNE-2 RNA A 260 / A 280 1. 89 ~ 1. 92 2 28 s18 s 2 1 RNA CNE- 2R CNE-2 3. CNE-2R CNE-2 CNE-2R CNE-2 3 3 46 000 CNE- 4. 1 CNE-2 CNE-2R 596 369 2R CNE-2 2 374 Kegg 227 2 870 Biocarta Toll IL-1R 485 385 2 2 374 13 2 2 Kegg Biocarta % Toll 11 2. 96-1 a EASE P a JUN MYD88 CCL5 CXCL10 STAT1 IL-1β LY96 FOS CCL3IL-6IL-8 5. 028 4. 956 10-5 6 1. 62 JUN MYD88 IRAK2IL-1βIL-1α IL-6 5. 610 0. 0026
170 2011 4 31 2 Chin J Radiol Med Prot April 2011 Vol. 31 No. 2 3 2 ID P 3725 JUN 28. 66238 0. 001531 Homo sapiens jun oncogene JUN mrna 4615 MYD88 26. 64086 0. 005467 Homo sapiens myeloid differentiation primary response gene 88 MYD88 mrna 6352 CCL5 121. 11240 0. 001294 Homo sapiens chemokine C-C motif ligand 5 CCL5 mrna 3627 CXCL10 25. 41306 0. 002646 Homo sapiens chemokine C-X-C motif ligand 10 CXCL10 mrna 6772 STAT1 81. 41203 0. 001953 Homo sapiens signal transducer and activator of transcription 191kDa STAT1 transcript variant alphamrna 23643 LY96 30. 72667 0. 003516 Homo sapiens lymphocyte antigen 96 LY96 mrna 2353 FOS - 64. 16970 0. 001287 Homo sapiens v-fos FBJ murine osteosarcoma viral oncogene homolog FOS mrna 6348 CCL3 64. 24246 0. 000226 Homo sapiens chemokine C-C motif ligand 3 CCL3 mrna 3576 IL-8 59. 40661 0. 001971 Homo sapiens interleukin 8 IL-8 mrna 3552 IL-1α 42. 98013 0. 000776 Homo sapiens interleukin 1alpha IL-1α mrna 3553 IL-1β 36. 21473 0. 000628 Homo sapiens interleukin 1beta IL-1β mrna 3569 IL-6 125. 29950 0. 000777 Homo sapiens interleukin 6 interferonbeta 2 IL-6 mrna 3656 IRAK2 31. 80981 0. 005798 Homosapiens interleukin-1 receptor-associated kinase 2 IRAK2 mrna Toll IL-1R 16-17 Legué 7 18 IL-6 3 Toll 1 TLR2 SC γ IL-6 TLR6 MYD88 TOLLIP TIRAP Efimova 19 FADD CASP8 2 NU61 LY96 TLR4 MYD88 TIRAP NF-κB IL- IL-6 IL-8 NU61 6 IL-1β 3 TLR5 TLR7 IL-6 TLR8 TLR9 MYD88 IRAK4 Hui 20 STAT1 NF-κB -1 AP-1 CNE-2R STAT1 IL-8 CCL5 CCL3 CXCL10 IL-6 IL-8 STAT1 IL-1R DNA IL-1α IL-1β 8 TOLLIP IRAK MYD88 JNK CNE-2R Toll P38 NF-κB c-jun IL-6 CASP8 c-jun AP-1 IL-1R NF-κB NF-κB AP-1 NF-κB NF-κB Toll IL-1R 9-14 -1 AP-1 15 CNE-2R CNE-2 Toll NF-κB AP-1 NF-κB AP-1 IL-6 IL-6 1. mirna.
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