SCGE、MNT与FCM对食管癌放疗患者外周血淋巴细胞损伤及不良反应的观察
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摘要
目的应用单细胞凝胶电泳法(Single Cell Gel Electrophoresis,SCGE)、微核试验(Micronucleus Test,MNT)和流式细胞技术(flow cytometry,FCM)分别检测放疗对食管癌患者外周血中淋巴细胞DNA损伤及T淋巴细胞亚群水平的变化情况,探讨放射剂量与外周血中淋巴细胞DNA损伤程度、疗效及不良反应的关系,为监测放疗对食管癌患者机体正常组织的损伤程度及细胞免疫功能的影响,探索更可靠的临床生物学量化指标。
方法采用SCGE、MNT技术测定36例食管癌放疗患者不同放疗剂量时外周血淋巴细胞DNA损伤及不良反应情况。同时应用FCM检测患者不同放疗剂量时T淋巴细胞亚群的变化。
结果①SCGE检测结果:正常对照组的外周血彗星拖尾率为18%,平均尾长为0.88±0.05;放疗前,患者组的外周血彗星拖尾率为22%,平均尾长为16.42±1.53;放疗剂量20Gy时,患者组的彗星拖尾率为44.6%,平均尾长为33.74±4.26;放疗剂量30Gy时,患者组的彗星拖尾率为50.5%,平均尾长为35.12±3.45;放疗剂量至40Gy时,患者组的彗星拖尾率为73.9%,平均尾长为41.83±2.36。②MNT检测结果:食管癌患者放疗前微核细胞率为0.31‰,放疗剂量20Gy时,微核细胞率为0.66‰;30 Gy时,微核细胞率为1.13‰;40Gy时,微核细胞率为2.31‰。③食管癌患者在确诊时,外周血T淋巴细胞亚群即存在细胞免疫功能的异常,表现为CD3~+和CD4~+以及CD4~+/CD8~+比例下降,而CD8~+变化不明显,而在接受放疗过程中,CD3~+和CD4~+以及CD4~+/CD8~+比例进一步下降。
结论①不同放疗剂量对食管癌患者外周血淋巴细胞DNA确实存在损伤作用,且损伤程度不同。②放疗使食管癌患者机体的免疫功能较放疗前有不同程度的下降。③不同的放疗剂量可引起食管癌患者不同程度的不良反应。④本研究初步表明,食管癌患者外周血淋巴细胞DNA损伤及T淋巴细胞亚群监测,对食管癌放疗疗效评价有一定的参考意义,因此有必要做进一步的深入探讨。
Objective:By using the methods of SCGE(Single Cell Gel Electrophoresis),MNT (Micronucleus Test),and FCM(Flow Cytometry)respectively,DNA damage,adverse effects and variation situation of T lymphocyte subgroups in the peripheral lymphocyte of esophaus tumor sufferers after radiotherapy are measured;and the causality between radiotherapy dose and DNA damage degree to normal histiocyte are studied.The objective is to measure the DNA damage degree to normal histiocyte,the influence to immune ability of body of esophagus tumor sufferers after the radiotherapy.By means of this effect evaluation,more reliable quantitative change on clinical biology is studied.
Methods:By using the methods of SCGE and MNT,DNA damage situation in the peripheral lymphocyte of 36 cases in different radiotherapy stages are measured. Meanwhile,by using the method of FCM,variation situation of the T lymphocyte groups of them in different stages are measured.
Results:1.As to the SCGE measurement,the rate of length migration of the peripheral lymphocyte in healthy groups was 18%,averaged tail length was 0.88±0.05;the rate of length migration in patient groups before radiotherapy was 22%,averaged tail length was16.42±1.53;when the radiation dose was 20Gy,the rate of length migration was 44.6 %,averaged tail length was33.74±4.26;when radiation dose amounted to 30 Gy,the rate of length migration was 50.5%,averaged tail length was35.12±3.45;when radiation dose reached to 40 Gy,the rate of length migration was73.9%,averaged tail length was 41.83±2.36.2.As to the MNT measurement,the micronucleus cell rate of this sample was 0.66‰before the radiotherapy,and was 1.81‰before the radiotherapy.3.As to the immune function measurement,when they are diagnosed the esophagus tumor sufferers, their immune function of T lymphocyte subgroups were a little abnormal,that was,the proportion of the CD3~+,CD4~+,andCD4~+/CD8~+ was decreased while the CD8~+ was normal;but during the radiotherapy the proportion of the CD3~+,CD4~+and CD4~+/CD8~+ was decreased more.
Conclusion:1.The different radiation dose can lead to the DNA damage of in the peripheral lymphocyte of the esophagus tumor patients;and the damaging degree varies according to difference of dose.2.Radiotherapy can reduce their immune function of the esophagus tumor sufferers in different degree.3.Different radiation dose may result in adverse effects in different degree.4.This study showed that the measurement to DNA damage of in the peripheral lymphocyte of the esophagus tumor sufferers,as well as that to T lymphocyte subgroups can provide some references on effect evaluation of radiotherapy to esophagus tumor.Therefore,more deeper discussion is necessary.
方法采用SCGE、MNT技术测定36例食管癌放疗患者不同放疗剂量时外周血淋巴细胞DNA损伤及不良反应情况。同时应用FCM检测患者不同放疗剂量时T淋巴细胞亚群的变化。
结果①SCGE检测结果:正常对照组的外周血彗星拖尾率为18%,平均尾长为0.88±0.05;放疗前,患者组的外周血彗星拖尾率为22%,平均尾长为16.42±1.53;放疗剂量20Gy时,患者组的彗星拖尾率为44.6%,平均尾长为33.74±4.26;放疗剂量30Gy时,患者组的彗星拖尾率为50.5%,平均尾长为35.12±3.45;放疗剂量至40Gy时,患者组的彗星拖尾率为73.9%,平均尾长为41.83±2.36。②MNT检测结果:食管癌患者放疗前微核细胞率为0.31‰,放疗剂量20Gy时,微核细胞率为0.66‰;30 Gy时,微核细胞率为1.13‰;40Gy时,微核细胞率为2.31‰。③食管癌患者在确诊时,外周血T淋巴细胞亚群即存在细胞免疫功能的异常,表现为CD3~+和CD4~+以及CD4~+/CD8~+比例下降,而CD8~+变化不明显,而在接受放疗过程中,CD3~+和CD4~+以及CD4~+/CD8~+比例进一步下降。
结论①不同放疗剂量对食管癌患者外周血淋巴细胞DNA确实存在损伤作用,且损伤程度不同。②放疗使食管癌患者机体的免疫功能较放疗前有不同程度的下降。③不同的放疗剂量可引起食管癌患者不同程度的不良反应。④本研究初步表明,食管癌患者外周血淋巴细胞DNA损伤及T淋巴细胞亚群监测,对食管癌放疗疗效评价有一定的参考意义,因此有必要做进一步的深入探讨。
Objective:By using the methods of SCGE(Single Cell Gel Electrophoresis),MNT (Micronucleus Test),and FCM(Flow Cytometry)respectively,DNA damage,adverse effects and variation situation of T lymphocyte subgroups in the peripheral lymphocyte of esophaus tumor sufferers after radiotherapy are measured;and the causality between radiotherapy dose and DNA damage degree to normal histiocyte are studied.The objective is to measure the DNA damage degree to normal histiocyte,the influence to immune ability of body of esophagus tumor sufferers after the radiotherapy.By means of this effect evaluation,more reliable quantitative change on clinical biology is studied.
Methods:By using the methods of SCGE and MNT,DNA damage situation in the peripheral lymphocyte of 36 cases in different radiotherapy stages are measured. Meanwhile,by using the method of FCM,variation situation of the T lymphocyte groups of them in different stages are measured.
Results:1.As to the SCGE measurement,the rate of length migration of the peripheral lymphocyte in healthy groups was 18%,averaged tail length was 0.88±0.05;the rate of length migration in patient groups before radiotherapy was 22%,averaged tail length was16.42±1.53;when the radiation dose was 20Gy,the rate of length migration was 44.6 %,averaged tail length was33.74±4.26;when radiation dose amounted to 30 Gy,the rate of length migration was 50.5%,averaged tail length was35.12±3.45;when radiation dose reached to 40 Gy,the rate of length migration was73.9%,averaged tail length was 41.83±2.36.2.As to the MNT measurement,the micronucleus cell rate of this sample was 0.66‰before the radiotherapy,and was 1.81‰before the radiotherapy.3.As to the immune function measurement,when they are diagnosed the esophagus tumor sufferers, their immune function of T lymphocyte subgroups were a little abnormal,that was,the proportion of the CD3~+,CD4~+,andCD4~+/CD8~+ was decreased while the CD8~+ was normal;but during the radiotherapy the proportion of the CD3~+,CD4~+and CD4~+/CD8~+ was decreased more.
Conclusion:1.The different radiation dose can lead to the DNA damage of in the peripheral lymphocyte of the esophagus tumor patients;and the damaging degree varies according to difference of dose.2.Radiotherapy can reduce their immune function of the esophagus tumor sufferers in different degree.3.Different radiation dose may result in adverse effects in different degree.4.This study showed that the measurement to DNA damage of in the peripheral lymphocyte of the esophagus tumor sufferers,as well as that to T lymphocyte subgroups can provide some references on effect evaluation of radiotherapy to esophagus tumor.Therefore,more deeper discussion is necessary.
引文
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