西妥昔单抗对人食管癌放射抗拒性细胞株KYSE-150R放射敏感性的研究

Objective To determine the effect of cetuximab(C225)on the radioresistant human esophageal squamous carcinoma eell line KYSE-150R.Methods A radioresistant human esophageal squamous carcinoma cell line KYSE-150R was established by fractionated irradiation.Morphological changes from KYSE-150 to KYSE-150R were observed by phase-contrast microscopy.Karyotype analysis was performed by G-banding.The radiosensitivities were analyzed by colony formation assays.Results The population doubling time of KYSE-150 and KYSE-150R were(23.6±0.2)h and(25.9±0.6)h (t=6.6,P<0.01),respectively.The chromosome number of KYSE-150R was increased and chromosome aberrations were observed from(69.3±1.9)h to(73.7±1.2)h(t=-8.83,P<0.01).The SF2,D0,Dq and N values of KYSE-150R were all higher than those of KYSE-150.After 5μg/ml of C225 added,the SF2,D0,Dq and N values were significantly decreased as compared to the control.After C225 treatment,the G0/G1 and G2/M phase cells were increased,while S-phase cells decreased(t=-4.478-4.308,P<0.05).Conclusion Cetuximab can enhance the radiosensitivity of radioresistant human esophageal squamous carcinoma cell line KYSE-150R.     

鼻咽癌放射抗拒相关的信号通路

Objective To study the difference of gene expression profile between the radioresistant human nasopharyngeal carcinoma cell line CNE-2R and CNE-2,and 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 60Co 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-1 R-mediated signal transduction pathway to the radioresistance of the CNE-2R cells and the significant differences of 13 genes in these 2 pathways,including JUN,MYD88,CCL5,CXCL10,STAT1,LY96,FOS,CCL3,IL-6,IL-8,IL-1α,IL-1B,and 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.     

沙利度胺对食管癌TE1细胞DNA合成抑制作用和辐射敏感性的影响

Objective To explore the radiosensitization effect of thalidomide combined with X-ray on esophageal carcinoma TE1 cells.Methods Cell scratch assay Was used to detect the inhibition ability of different concentration of Thalidomide on cell invasion and metastasis.H3-TdR incorporation assay Was used to investigate the inhibition of DNA synthesis in TE1 cells by treated with Thalidomide singly or combination with X-rays.The colony formation assay Was used to analyze the radiosensitization of Thalidomide effect on TE1 cells.Results Thalidomide had obvious inhibition effect on TE1 cell metastasis.DNA synthesis and colony formation,which were correlated with drug concentration.The values D0,Dq and SF2 in TE1 cells were gradually decreased with thalidomide concentration increased.When the concentration of thalidomide was 100μg/ml,the SERD0 and SERDq were(1.4±0.2)and(1.5±0.1),respectively,While the concentration of thalidomide Was 1 50μg/ml,the SERD0 and SERDq were metastasis,DNA synthesis,and significantly enhance the radiosensitizing effect on esophageal carcinoma TE1 cells.     

沙利度胺对食管癌TE1细胞DNA合成抑制作用和辐射敏感性的影响

Objective To explore the radiosensitization effect of thalidomide combined with X-ray on esophageal carcinoma TE1 cells.Methods Cell scratch assay Was used to detect the inhibition ability of different concentration of Thalidomide on cell invasion and metastasis.H3-TdR incorporation assay Was used to investigate the inhibition of DNA synthesis in TE1 cells by treated with Thalidomide singly or combination with X-rays.The colony formation assay Was used to analyze the radiosensitization of Thalidomide effect on TE1 cells.Results Thalidomide had obvious inhibition effect on TE1 cell metastasis.DNA synthesis and colony formation,which were correlated with drug concentration.The values D0,Dq and SF2 in TE1 cells were gradually decreased with thalidomide concentration increased.When the concentration of thalidomide was 100μg/ml,the SERD0 and SERDq were(1.4±0.2)and(1.5±0.1),respectively,While the concentration of thalidomide Was 1 50μg/ml,the SERD0 and SERDq were metastasis,DNA synthesis,and significantly enhance the radiosensitizing effect on esophageal carcinoma TE1 cells.     

Dose-effect of chromosome aberration in lymphocytes of human peripheral blood irradiated with high dose of 60Co γ rays in vitro

Objective To explore the dose-effect of chromosome aberration,and establish a dose-effect curve of dicentrie and ring (die+r) in human peripheral blood lymphocytes irradiated with high dose of γ-rays.Methods Human peripheral blood was obtained from three healthy individuals,and exposed to 60Co γ-rays with doses between 0 and 50 Gy.Colchicine was added immediately and the cells were incubated at 37℃ for 52,72 and 96 h,respectively.Metaphase index (MI) and dic+r were counted,and a dose-effect curve was fitted.Dose estimation were performed according to the curve for the two patients who had accidentally received high doses of irradiation.Results MI was reduced with the dose increasing.Frequencies of dic+r were increased until 23 Gy,and then saturation was observed.A dose-effect curve was fitted during 5-23 Gy,and the doseeffect was as follows:γ=-1.608(±0.300)+0.830(±0.051) D-0.013(±0.002) D2(R2=0.998),where γ is the yield of dic+r,and D is the dose.The dose estimated for the two patients were in accordance with those by physical method,ESR method and clinical symptoms.Conclusions The highest dose that could be estimated is 23 Gy with the established dic+r dose-effect curve by using the eolchicine black method and prolonged culture time,which suggests the potential of chromosome aberration analysis as biological dosemeter.     

塞来昔布对人肺腺癌细胞株A549的放射增敏效应及细胞迁移力的影响

Objective To investigate the effects of radiosensitivity enhancement and inhibition of migration ability of human lung adenocarcinoma cells by celecoxib,a selective cyclooxygenase (COX)-2 inhibitor.Methods Human lung adenocarcinoma cells of the line A549 were cultured and then inoculated into six-well plates and randomly divided into 4 groups:control group,celecoxib group administered with celecoxib at the subtoxic doses 30 and 50 μmol/L,irradiated group exposed to 0,1,2,4,6,or 8 Gy by linear accelerator,and combined treatment (celecoxib + irradiation) group.The radiosensitizing effect of celecoxib was assessed by clonogenic cell survival test.The migration ability of the A549 cells was measured by scratch-wound test and the content of metalloproteinase-2 (MMP-2) in culture supernatant was detected with ELISA.Results The sensitization enhancement ratio of the celexib group was increased dosedependently.The values of D0 ,Dq,SF2 and D0.01 of the celecoxib + irradiation group were all significantly lower than those of the irradiated group.Scratch-wound test showed that the no-scratch area of the celecoxib + irradiation group and celecoxib group were all significantly wider than those of the mere irradiation and control groups and there was a dose-dependent manner,and the no-scratch area of the celecoxib + irradiation group was wlider than that of the celecoxib group.ELISA showed that the MMP-2 levels in the supernatant of the celecoxib group and celecoxib + irradiation group were respectively significantly lower than those of the control group and mere irradiated group (t = 3.78,5.79、3.15,P ＜ 0.05),however,there was not significant difference between the mere irradiation and control groups (t = 2.73,2.38,P ＞ 0.05).Conclusions Celecoxib enhances concentration-dependently the radiosensitivity of human lung carcinoma cell and inhibits the secretion of MMP-2 of the carcinoma cells,thus inhibiting their migration ability.     

食管胸中段癌三维适形放疗中呼吸运动导致靶区移位的研究

Objective To evaluate the respiration-induced target volume motion in 3D-CRT for mid-thoracic esophageal carcinoma in order to guide the radiation oncologist to choose the expansion margin for ITV.Methods Ten patients with mid-thoracic esophageal carcinoma were scanned by multi-spiral CT simulator respectively in free breathing(FB),breath.hold after normal inspiration and expiration(IBH and EBH)with the same scanning range.Then the CT images of three series were transfefred to the treatment planning system.The target volume was outlined following the same standard.The motion of the center point of GTV,the center point of each slice of GTV and the edge of the GTV in selected slice were measured respectively to obtain the comprehensive value of GTV motion。in order to find the appropriate IM value according to the 95％confidence interval of the GTV motion.Results ①The GTV motion between IBH and EBH was(0.19±0.16)cm in the left.right direction,(0.54±0.19)cm in the cranial and caudal irection.and(0.16±0.14)cm in anterior.posterior directions for the center of GTV,.For the center point of each slice of GTV.they ere(0.19±0.15)cm,(0.54±0.16)cm,(0.16±0.13)cm in three directions above.respectively.For the edge of the GTV in selected slice.they were(0.26±0.19)cm,(0.54±0.18)cm,(0.24±0.19)cm,respectively.The comprehensive value of GTV motion between IBH and EBH was(0.23±0.17)cm,(0.54±0.17)cm,(0.21±0.17)cm.respectively.The 95％confidence interval was 0.21-0.25 cm.0.53-0.56 cm and 0.19-0.22 cm in three directions.②The direction of GTV motion：No motion was noticed in 8.2％.while 73.3％to the right side and 18.5％to the left side in the left-right direction when IBH were compared with EBH.100％were moved to caudal in the the cranial and caudal direction[(0.54±0.17)cm].In the anterior-posterior direction,no motion was noticed in 8.2％,while 16.6％to the posterior and 75.2％to the anterior when IBH were compared with EBH.③The GTV motion was correlated with the vafiance of 1ung volumes in IBH-EBH(r=0.683,P=0.032)and not with GTV volume and length.Conclusions Respiration can induce target volume motion in 3 DCRT for mid-thoracic esophageal carcinoma.Compared to EBH.the GTV tends to move to the caudal,the anterior and the ight side in IBH.     

食管胸中段癌三维适形放疗中呼吸运动导致靶区移位的研究

Objective To evaluate the respiration-induced target volume motion in 3D-CRT for mid-thoracic esophageal carcinoma in order to guide the radiation oncologist to choose the expansion margin for ITV.Methods Ten patients with mid-thoracic esophageal carcinoma were scanned by multi-spiral CT simulator respectively in free breathing(FB),breath.hold after normal inspiration and expiration(IBH and EBH)with the same scanning range.Then the CT images of three series were transfefred to the treatment planning system.The target volume was outlined following the same standard.The motion of the center point of GTV,the center point of each slice of GTV and the edge of the GTV in selected slice were measured respectively to obtain the comprehensive value of GTV motion。in order to find the appropriate IM value according to the 95％confidence interval of the GTV motion.Results ①The GTV motion between IBH and EBH was(0.19±0.16)cm in the left.right direction,(0.54±0.19)cm in the cranial and caudal irection.and(0.16±0.14)cm in anterior.posterior directions for the center of GTV,.For the center point of each slice of GTV.they ere(0.19±0.15)cm,(0.54±0.16)cm,(0.16±0.13)cm in three directions above.respectively.For the edge of the GTV in selected slice.they were(0.26±0.19)cm,(0.54±0.18)cm,(0.24±0.19)cm,respectively.The comprehensive value of GTV motion between IBH and EBH was(0.23±0.17)cm,(0.54±0.17)cm,(0.21±0.17)cm.respectively.The 95％confidence interval was 0.21-0.25 cm.0.53-0.56 cm and 0.19-0.22 cm in three directions.②The direction of GTV motion：No motion was noticed in 8.2％.while 73.3％to the right side and 18.5％to the left side in the left-right direction when IBH were compared with EBH.100％were moved to caudal in the the cranial and caudal direction[(0.54±0.17)cm].In the anterior-posterior direction,no motion was noticed in 8.2％,while 16.6％to the posterior and 75.2％to the anterior when IBH were compared with EBH.③The GTV motion was correlated with the vafiance of 1ung volumes in IBH-EBH(r=0.683,P=0.032)and not with GTV volume and length.Conclusions Respiration can induce target volume motion in 3 DCRT for mid-thoracic esophageal carcinoma.Compared to EBH.the GTV tends to move to the caudal,the anterior and the ight side in IBH.