鼻咽癌放射抗拒相关的信号通路

目的 研究具有不同放射抗拒性的人鼻咽癌细胞株CNE-2R与CNE-2的基因表达谱差异,筛选与鼻咽癌放射抗拒相关的基因信号通路.方法 验证已构建的鼻咽癌放射抗拒细胞株CNE-2R的放射抗拒性,选用Human-6v3.0全基因组表达谱微珠芯片,筛选这2种细胞的差异表达基因.对差异基因进行生物信息学分析,寻找与鼻咽癌放射抗拒...     

放射性肺炎

在胸部恶性肿瘤的放射治疗中 ,肺组织会受到不同程度的照射。当肺组织接受较高剂量和 /或较大容积的照射 ,或患者对放射线敏感时 ,可能发生放射性肺炎。1　发病率及病因〔1〕　　有症状的放射肺炎的发病率为 5 %～ 15 %。发病与下列因素有关 :(1)受照射肺的容积 :剂量相同时受照射肺组织容积越大 ,发生率越高。 (2 )受照射剂量 :放射剂量 <15Ｇｙ时 ,很少发生放射性肺炎 ;若 >6 0Ｇｙ会发生不同程度的放射性肺炎。 (3)分割方式 :总剂量相同时 ,分割次数越少 ,总疗程越短 ,放射性肺炎的发病率越高。 (4)受照射部位 :上肺及近纵隔的肺组织较下…     

肿瘤放射增敏药物的研究进展

放射治疗是临床肿瘤治疗的重要手段之一.肿瘤细胞的放射敏感性是影响肿瘤放疗疗效的关键因素.放射增敏药物能够增强机体的放射敏感性,通过提高肿瘤细胞的放射敏感性达到降低照射剂量、提高放疗疗效、降低正常组织损伤的目的.现有的放射增敏药物主要分为细胞毒类药物、靶向药物以及中药制剂3大类.该文将对肿瘤放射增敏药物的作用机制、现状及相关研究进展进行综述.     

中性单细胞电泳检测肿瘤细胞放射敏感性

目的　探讨中性单细胞电泳检测肿瘤细胞放射敏感性的可行性。方法　应用克隆形成法和中性单细胞电泳技术 ,检测辐射诱导的人红白血病细胞株K562 、人结肠腺癌细胞株LS T 1 1 7和鼠胶质瘤细胞株C6的初始DNA双链断裂数及双链断裂后的修复 ,以及两者与细胞放射敏感性之间的关系。结果　 3种细胞系的放射敏感性依次为K562 >LS T 1 1 7>C6;3种细胞系的DNA迁移长度都随着照射剂量的增加而增大 ,呈良好的剂量效应关系。在相同剂量下辐射诱导的DNA双链的初始断裂数目 ,也依次为K562 >LS T 1 1 7>C6;3种细胞系经 1 0GyX射线照射 ,并在PBS中培养不同时间后DNA迁移长度 ,都有较大幅度的下降 ,但下降幅度依次为C6>LS T 1 1 7>K562 。在相同剂量下辐射诱导的DNA双链断裂后的修复能力 ,也依次为C6>LS T 1 1 7>K562 。结论　辐射诱导的DNA双链断裂及修复 ,与细胞放射敏感性有很好的相关性 ,可望用于肿瘤细胞内在放射敏感性的预测     

原位缺口平移测定鼻咽癌细胞的放射敏感性

报道了以原位缺口平移（ＩＳＮＴ）技术研究鼻咽癌细胞株ＣＮＥ、ＣＮＥ－２Ｚ对γ射线的放射效应，以及用存活曲线验证ＩＳＮＴ的可靠性。结果表明，在０～８Ｇｙ剂量之间，ＣＮＥ及ＣＮＥ－２Ｚ的脱氧三磷酸核苷（ｄＮＴＰ）掺入率与照射剂量间呈良好量效关系；ＣＮＥ－２Ｚ的放射敏感性高于ＣＮＥ。相关分析表明，ｃＮＥ及ＣＮＥ－２Ｚ细胞ＩＳＮＴ的ｄＮＴＰ掺入率与存活分数之间均有良好的相关关系。表明ＩＳＮＴ可以较好地反映射线对肿瘤细胞的损伤效应，可替代复杂的集落形成试验，可望成为评价肿瘤细胞放射敏感性的一个重要指标。     

中枢神经系统的放射损伤及其处理

系统综述中枢神经系统放射损伤的病理生理改变、临床表现、诊断标准及防治措施。指出中枢神经系统放射损伤与受照射体积、分次剂量、分割次数及照射总剂量密切相关,放疗与化疗同时进行将增加脑、脊髓急性放射损伤的发生率。强调减少中枢神经系统放射损伤并发症的关键在于预防。     

鳞癌细胞株放射敏感性与端粒酶活性及端粒长度变化的关系

目的探索放射抗拒鳞癌细胞株放射敏感性的改变与端粒酶活性、端粒长度变化的关系。方法以人喉鳞癌细胞株Hep2为实验对象,用γ射线反复照射获得具有放射抗拒性细胞株Hep2R,拟合细胞存活曲线比较两种细胞株的放射生物学参数及端粒酶抑制剂AZT(azidothymidine,叠氮胸苷)作用后的变化,用TRAP-ELISA法测定端粒酶活性(OD值),Southernblotting法分析端粒平均长度(TRF),比较端粒酶活性、端粒平均长度的变化。结果辐射诱导出一个具有放射抗拒性的Hep-2R细胞株,并已稳定传代培养30代以上且放射抗拒性能稳定,测得其SF2=0.6798、D0=3.24,Hep-2R细胞的端粒酶活性较Hep-2细胞升高(0.982±0.005vs0.604±0.015),端粒长度延长了2倍(11.12kbvs3.76kb),AZT作用后Hep2R细胞株SF2=0.4892、D0=2.51,端粒酶活性下降为0.708±0.011、端粒长度缩短为10.18kb。Hep2细胞对应的参数SF2=0.4148、D0=2.06,AZT作用后Hep2细胞SF2=0.3843、D0=1.81,端粒酶活性下降为0.364±0.003、端粒长度缩短为2.76kb。结论辐射诱导细胞获放射抗拒性后其端粒酶活性升高、端粒平均长度增长,端粒酶抑制剂能通过降低端粒酶活性、缩短端粒长度而对其有增敏效应。     

中枢神经系统的放射损伤及其处理

系统综述中枢神经系统放射损伤的病理生理改变,临床表现,诊断标准及防治措施,指出中枢神经系统放射损伤与受照射体积,分次剂量,分割次数及照射总剂量密切相关,放疗与化疗同时进行将增加脑,脊髓急性放射损伤的发生率,强调减少中枢神经系统放射损伤并发症的关键在于预防。     

由SABR再论放射敏感性

随着现代放射治疗技术进步,放疗已由过去二维时代进入三维和四维时代,治疗精度大幅度提高,分割模式也发生了深刻变革。从传统放射治疗发展到以三维适形放射治疗（3D-CRT）和调强放射治疗（IMRT）为代表的聚焦照射,提高了肿瘤靶区剂量,减少了正常组织的损伤。同时随着影像引导技术进步,治疗机与影像引导结合,每次治疗前通过影像扫描技术获得肿瘤靶区位置信息,或用4D影像引导技术精确地将射线投射到目标靶点,达到立体定向体部放射治疗（stereotactic body radiation therapy,SBRT）/立体定向消融放疗（stereotactic ablative radiotherapy,SABR）的目的,放射治疗完全进入精准、高效和低毒时代。高剂量、大分割照射已经取得令人信服和可喜的疗效,传统放射生物学理论已无法解释这种照射模式抗肿瘤细胞作用机制。传统放疗认为,肿瘤有敏感与不敏感之分,但是,进入SABR时代,肿瘤对其治疗均反应良好,放射治疗学迫切需要建立新的放射生物学学说和体系,在传统放射生物学理论基础上,更好地阐明新技术原理、作用机制,并建立与传统放射生物学内在联系,为临床普及和推广消融放疗技术奠定理论基础。     

人食管癌细胞株EC-9706照射后细胞周期的研究

放射牛物学研究表明,电离辐射作用后,不同肿瘤细胞的细胞周期变化不同.本研究主要采用流式细胞仪法检测人食管癌细胞株Ec-9706接受不同剂量6 MV X射线照射后细胞周期的变化情况.     

Targeting the AKT/cyclin D1 pathway to overcome intrinsic and acquired radioresistance of tumors for effective radiotherapy

Purpose: Radiotherapy (RT) is a powerful tool in the treatment of cancer, having the advantage of preserving normal tissues. Clinical outcomes of RT are significantly improved by technological advances, enabling increased radiation doses directed very specifically to a tumor. However, tumor radioresistance remains a major impediment to effective RT. We have shown that human tumor cells surviving after repeated exposure to fractionated radiation (FR) of X-rays for 1 month have acquired radioresistance through constitutive activation of AKT and downstream cyclin D1 nuclear retention. Tumor radioresistance is also proposed to be an intrinsic characteristic of cancer stem cells (CSC), whose efficient DNA repair is thought to confer this phenotype. We have isolated radioresistant CD133-positive cells following exposure to long-term FR. These cells exhibited the CSC phenotype with activation of the AKT/cyclin D1 pathway. In this review, I summarize our current understanding of the molecular mechanisms underlying tumor radioresistance and propose a strategy for overcoming radioresistance by targeting the AKT/cyclin D1 pathway.

Conclusion: Two different mechanisms: acquired radioresistance of surviving tumor cells after RT and intrinsic radioresistance of CSC are associated with tumor radioresistance. Inhibition of the AKT pathway results in radiosensitization of both types of tumor radioresistance.     

辐射相关基因芯片的制备及其在肺癌细胞中的应用

目的 寻找参与肺癌细胞辐射抗性的基因。方法构建辐射相关的基因芯片，初步筛选出在不同辐射敏感性肺癌细胞系中差异表达的基因。为了评价芯片结果的可靠性，我们对一些差异表达基因进行了RT-PCR验证。结果和辐射敏感的NCI-H446细胞相比，辐射抗性的A549中有18个基因有明显的改变，8个基因是上调，10个基因是下调。在照射后6h和24h，A549细胞中分别有22个基因(19个基因上调，3个下调)和26个基因(8个上调，18个下调)的差异表达；NCI-H446细胞中分别有17个基因(9个基因上调，8个下调)和18个基因(6个上调，12个下调)差异表达。从这些基因中，我们发现一些参与细胞增殖和抗凋亡的基因，在照射后的A549细胞中是增高的，而在NCI-H446细胞中是降低的。另外一些参与DNA修复的基因，在A549中比在NCI-H446细胞中有更高的表达。结论一些参与细胞DNA修复、细胞周期调控、细胞增殖和抗凋亡作用的基因可能有助于A549细胞辐射抗性的形成。     

Stable radioresistance in ataxia-telangiectasia cells containing DNA from normal human cells

SV40-transformed ataxia-telangiectasia (AT) cells were transfected with a cosmid that contains a normal human DNA library and a selectable marker, the neo gene, which endows successfully transformed mammalian cells with resistance to the antibiotic G418. After a three-part selection protocol for G418 resistance and radioresistance, a cell line stably resistant to ionizing radiation was recovered. Cells from this line were irradiated with 50 Gy of X-rays and fused with non-transfected AT cells. Among the G418-resistant colonies recovered was one that was stably resistant to radiation. Resistance to ionizing radiation of both the primary transfectant line and its fusion derivative was intermediate between that of AT cells and normal cells, as assayed by colony-forming ability and measurement of radiation-induced G2 chromatid aberrations; both cell lines retained AT-like radioresistant DNA synthesis. These results suggest that, because radioresistance in the transfected cells was not as great as that in normal human cells, the two hallmarks of AT, radiosensitivity and radioresistant DNA synthesis, may still be the result of a single defective AT gene.     

食管癌细胞系TE13R120放射抗性与HDAC3、NF-kB和NRAGE关系的研究

目的探讨HDAC3、NF—kB和NRAGE基因与食管癌细胞放射抗性的关系。方法以人食管癌细胞系TE13和由TE13经γ射线反复照射建立的放射抗性细胞系TE13R120为研究对象，用Western blotting技术检测这两种细胞于不同时间、剂量点照射后基因HDAC3、NF—kB和NRAGE的蛋白表达；用流式细胞术检测相同时间、剂量点两种细胞的凋亡和细胞周期分布情况。结果Western blotting显示，与TE13相比，TE13R120细胞照射前后HDAC3的核表达均增高，NF-kB的表达也明显增强，NRAGE在两种细胞胞浆中的表达无差异。与TE13相比，TE13R120中S期细胞明显增多，照射后G2期阻滞明显，凋亡水平较低。结论基因HDAC3和NF-kB可能在食管癌细胞放射抗性形成中发挥作用并可能有协同作用；NRAGE与食管癌辐射抗性的关系有待进一步研究。     

Adaptive response of a new radioresistant strain of Chlamydomonas reinhardtii and correlation with increased DNA double-strandbreak rejoining

In order to study the relationship between radioresistance and the adaptive response, we aimed to produce a new strain of Chlamydomonas reinhardtii with characteristics of high radioresistance coupled with a protoplast structure typical for the genus, and the cell-wall-less phenotype to facilitate rapid cell lysis in DNA double-strand break (DSB) assays. The adaptive response of the new strain was investigated using clonogenic and DSB assays. Strain H-3 was derived by mating a radioresistant strain (AK-9-9) with the cell-wall-less mutant CW15 strain and selecting for radioresistance by clonogenic assay. The random amplification of polymorphic DNA (RAPD) molecular marker system was used to evaluate genetic polymorphisms between H-3 and other related C. reinhardtii strains. DSB were estimated using constant-field electrophoresis. Of several mutant strains tested, strain H-3 was shown to be most radioresistant on the basis of dose to give a 90% lethality (LD90) rate and dose to give a 99% lethality rate (LD99). In addition to its high radioresistance and thinner cell wall as compared with that of the other parental strain AK-9-9, H-3 also expressed a radiation-induced adaptive response measured by clonal survival when given a priming dose before a test dose. DSB were also rejoined more rapidly in cells exposed to a priming dose 4 h previously. It is concluded from split-dose experiments that the already highly radioresistant strain H-3 is further capable of 'over recovery' or adaptation to radiation exposure. Accelerated DSB rejoining in cells given a priming dose may underlie the cellular adaptive response in this organism.     

Adaptive responses in human glioma cells assessed by clonogenic survival and DNA strand break analysis

PURPOSE: Human gliomas are known to be radioresistant and the aim was to determine if this resistance in part could be due to an adaptive response. MATERIALS AND METHODS: Human U-87MG glioma cells were used. Three different radiation regimens that could be related to clinical treatments were tested for their ability to cause an adaptive response. Cell survival and DNA double-strand breakage were the measured endpoints. RESULTS: All three regimens caused an adaptive response in terms of cell survival when given priming doses of radiation. The DNA double-strand break endpoint also showed fewer breaks when the adaptive response occurred. CONCLUSIONS: Using irradiation regimens that closely resembled clinical applications, in vitro data are presented that show an adaptive response in human glioma cells. This effect in part could be responsible for the radioresistance of human gliomas.     

Measurement of oxygen concentration in tumor cells by fluorescent probe (nitrocompound)

The hypoxic cell fraction in tumors is considered to be responsible for radioresistance. Estimating the population of the hypoxic cell fraction in tumor could develop the effective means to predict radiosensitivity. In this study, nitroacridine (fluorescent dye) was tested to estimate hypoxic status in single cells and in spheroids. The oxygen concentration in the medium was measured by oxygen electrodes utilizing polarography method, mean while that in cells was calculated from the fluorescence intensity of the dye. The fluorescent spectra from cells showed the same pattern in spite of the changed oxygen concentration in medium, on the other hand its intensity was dependent upon the oxygen concentration. Using a fixed nitroacridine concentration and a fixed staining time, oxygen concentration of cells could be determined within range from 0.1 to 1.0% values. These values are almost the same as the oxygen concentration of the radioresistant tumor cells. Thus, the fluorescent method we used in this study is considered to be useful to estimate radioresistance of tumor. However, fluorescent intensity would alter when used different cell lines, because of different cellular activity of nitroreductase.     

Caveolin-1: a novel prognostic biomarker of radioresistance in cancer

Purpose: Caveolin-1 is a membrane protein highly expressed in many tumors and plays an important role in tumor progression and metastasis. This review describes the structure of the Caveolin-1 protein and its pre-clinical and clinical significance, demonstrating that Caveolin-1 is a novel biomarker for radioresistance which has the promising potential to improve the clinical outcome of cancer patients undergoing radiation treatment.

Summary: Targeted radiation therapy has shown immense benefits for cancer treatment. However, one of the major challenges for effective clinical outcome of radiation therapy for cancer patients is the development of radioresistance during radiation treatment. As a consequence, radiation therapy becomes a less effective modality for successful clinical outcome. Furthermore, a radioresistant tumor has the ability to repair its genome, and therefore becomes more aggressive and metastasizes. The plausible mechanisms for tumor radioresistance include the rapid DNA repair, somatic mutations in tumor oncogenes, aberrant activation of kinase pathways, and changes in the tumor microenvironment including tumor hypoxia, tumor vasculature, and cancer stem cells. Caveolin-1 is significantly upregulated in certain cancer cells and aberrantly mediates downstream signaling mechanisms. Notably, numerous recent research reports have shown the role of Caveolin-1 in tumor radioresistance and poor treatment outcome. Thus, Caveolin-1 could be a novel prognostic biomarker to monitor tumor radioresistance in cancer patients undergoing radiation therapy.

Conclusions: Caveolin-1 has the promising potential to become a novel prognostic biomarker to monitor tumor radioresistance and radiation response specifically in the prostate, pancreas, and lung cancer.     

Adaptive response of a new radioresistant strain of Chlamydomonas reinhardtii and correlation with increased DNA double-strandbreak rejoining

In order to study the relationship between radioresistance and the adaptive response, we aimed to produce a new strain of Chlamydomonas reinhardtii with characteristics of high radioresistance coupled with a protoplast structure typical for the genus, and the cell-wall-less phenotype to facilitate rapid cell lysis in DNA double-strand break (DSB) assays. The adaptive response of the new strain was investigated using clonogenic and DSB assays. Strain H-3 was derived by mating a radioresistant strain (AK-9-9) with the cell-wall-less mutant CW15 strain and selecting for radioresistance by clonogenic assay. The random amplification of polymorphic DNA (RAPD) molecular marker system was used to evaluate genetic polymorphisms between H-3 and other related C. reinhardtii strains. DSB were estimated using constant-field electrophoresis. Of several mutant strains tested, strain H-3 was shown to be most radioresistant on the basis of dose to give a 90% lethality (LD₉₀) rate and dose to give a 99% lethality rate (LD₉₉). In addition to its high radioresistance and thinner cell wall as compared with that of the other parental strain AK-9-9, H-3 also expressed a radiation-induced adaptive response measured by clonal survival when given a priming dose before a test dose. DSB were also rejoined more rapidly in cells exposed to a priming dose 4 h previously. It is concluded from split-dose experiments that the already highly radioresistant strain H-3 is further capable of ‘over recovery’ or adaptation to radiation exposure. Accelerated DSB rejoining in cells given a priming dose may underlie the cellular adaptive response in this organism.     

Radiation-induced effects on telomerase in gynecological cancer cell lines with different radiosensitivity and repair capacity

PURPOSE: Telomerase activation in response to irradiation might enhance the radioresistance of cells. Thus, we have investigated radiation-induced effects on telomerase in six gynecological cancer cell lines, with different intrinsic radiosensitivity and capacity for sublethal damage repair (SLDR). MATERIALS AND METHODS: Three endometrial adenocarcinoma (UM-EC-1, UT-EC-2B and UT-EC-3) and three vulvar squamous cell carcinoma (A431, UM-SCV-2 and UM-SCV-7) cell lines were irradiated with doses of 5, 10 and 25 Gy and the effects on telomerase were evaluated at 0.5, 6, 24 and 48 h post-irradiation. Telomerase activity was quantitatively measured by SYBR Green real-time telomeric repeat amplification protocol. RESULTS: The most radioresistant cell line A431 had the strongest stimulatory effects (approximately 2.0 - 2.5-fold) on telomerase activity 24 and 48 h post-irradiation with the highest radiation doses. In contrast to that, telomerase activities in the highly radiosensitive cell line UT-EC-2B remained below the basal level throughout the 48-h period of post-irradiation with the highest doses, and even a decline to approximately 50% of the basal level was found 24 h after exposure. In other cell lines being either moderately or highly radiation resistant, telomerase activity levels in response to irradiation remained mainly at the basal level or gradually increased. CONCLUSIONS: The present findings indicate that there might be a connection between the radiation-induced telomerase response and radiosensitivity. However, no correlation was found between the radiation-induced effects on telomerase and the sublethal damage repair capacity of the cells.