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1.
P53及其相关蛋白对X射线照射肝癌细胞周期的调节   总被引:1,自引:0,他引:1  
X射线照射人肝癌细胞HepG2, 照射后细胞存活随照射剂量增大明显下降。 流式细胞术分析, 不同剂量组照射后24 h均发生G2期阻滞。 照射后不同时间组的细胞周期分布也有不同, 照射后12 h, 有显著的S期延迟。 Western Blot 显示照射后24 h P53, MDM2, P21蛋白表达上升, 并有时间效应: P53在照射后24 h之内始终维持较高表达, MDM2和P21分别在照射后6和12 h的表达最高。 X射线照射通过影响P53及其相关蛋白的表达影响细胞周期。 HepG2 cells were irradiated with X ray at the doses of 0, 1.0, 2.0, 4.0 or 8.0 Gy and separately maintained in DMEM at 37 ℃ for 0, 6, 12 or 24 h. Colony forming assay showed that cell survival decreased with the irradiation dose increasing. Cell cycle was detected by FACS, the arrest of S phase was found after 12 h irradiation and arrest of G2 phase took place at 24 h after all irradiation doses, which suggested that cell cycle distribution was different in groups gathered after different maintaining time. The results of Western blotting showed that the expression of P53, MDM2 and P21 increased more after irradiation than the control. The expression of P53 remained high at 24 h after irradiation, while the levels of MDM2 or P21 arrived at the highest at 6 h or 12 h after irradiation respectively. The expressions of P21 after irradiation were in corresponding with the cell cycle distribution in the groups of different maintaining time. In conclusion, irradiation change the distribution of cell cycle by effecting the expression of P53 and its related proteins.  相似文献   

2.
选取对数生长期人肺癌细胞A549接受0—6.0 Gy 碳离子照射, 用克隆形成法检测细胞的存活率; 并于照射后12和24 h收集细胞, 用流式细胞术检测细胞周期各时相的细胞百分比, 观察不同剂量碳离子辐照对A549细胞周期进程的影响。 结果显示: 0—6.0 Gy 碳离子照射后细胞存活率显著下降; 照射后12 h细胞发生G0/G1期阻滞, 而照射后24 h, 1.0 Gy 照射组细胞在G0/G1期阻滞, 2.0—6.0 Gy 照射组细胞在G2/M期阻滞。 上述结果表明, 在A549细胞接受碳离子照射后的12 和24 h内, 1.0 Gy 照射可持续激活细胞G1期检查点, 而2.0—6.0 Gy 碳离子照射后其细胞周期进程是随时间变化的。 To investigate the effects of cell cycle progression of A549 cell induced by 12C6+ ion irradiation at different doses, the survival fractions of the A549 cells were determined by colony forming assay; cell cycles were analyzed by FACS at 12 h or 24 h after irradiation. The results showed that the percentage of survival in the A549 cells decreased with irradiation doses. Compared with control group, the percentage of the cells in G0/G1 phase significantly increased at 12 h after irradiation with different doses of 12C6+ ions. However, at 24 h after irradiation the percentage of the cells in G0/G1 phase significantly increased with 1.0 Gy 12C6+ ions, while the cells showed increasing percentage in G2/M phase with 2.0, 4.0 or 6.0 Gy 12C6+ ions. The results suggested that G1 cell cycle checkpoint was activated in 12—24 h after irradiation with 1.0 Gy 12C6+ ions, but after irradiation with 2.0—6.0 Gy 12C6+ ions, the cell cycle progression of the A549 cells changed with time.  相似文献   

3.
以低剂量γ射线(0.05 Gy)预照射人肝癌细胞hep G2, 8 h后再用高剂量(3 Gy)照射, 测定了细胞的克隆存活率和细胞周期。 结果表明, 低剂量辐射预处理可诱导hep G2细胞产生克隆存活适应性反应, 并且有助于细胞通过G2/M期阻滞; 低剂量辐射诱导的克隆存活适应性反应与增强的通过细胞周期阻滞的能力之间有一定的相关性。 Human hepatoma cells hep G2 were irradiated with 3 Gy of γ ray 8 hours after primed with 0.05 Gy of γ ray, thereafter,cell survival and cell cycle were determined. The results indicated that both survival adaptive response and the enhanced ability to overcome G2/M arrest could be induced by pre irradiation with low dose of γ ray. It is suggested that there is a certain correlation between the survival adaptive response and the enhanced ability to overcome cell cycle arrest.  相似文献   

4.
采用4 Gy X射线辐照人乳腺癌细胞MCF 7, 分别在照射后0, 2, 4, 8, 16, 24, 48, 72和144 h时间点收集细胞, 提取DNA并以聚合酶链式反应进行扩增, 再以BsaXI限制性内切酶对扩增产物进行消化, 消化产物以琼脂糖凝胶电泳进行分离。 结果表明, 4 Gy X射线辐照可引起D310片段突变, 且该突变在经辐照之后144 h检测水平明显超过野生型。 Human breast cancer cell line MCF 7 was irradiated with 4 Gy X ray, collected at 0, 2, 4, 8, 16, 24, 48, 72, 144 h after irradiation, respectively. Whole genome DNA including mtDNA were extracted at each time point, and amplified by polymerase chain reaction(PCR). Then the PCR product was subjected to BSAXI digestion, all of digestion product then underwent a brief electrophoresis. Results showed D310 mutation can be induced by 4 Gy X ray irradiation and D310 mutation can overwhelm the normal phenotype 144 h after irradiation.  相似文献   

5.
本研究旨在探讨羧甲基-β-1,3葡聚糖(CMG)对人肝癌HepG2细胞X射线或12C6+离子束辐射敏感性的影响。首先用CCK-8法检测CMG对HepG2细胞的生长抑制情况,得到半数抑制浓度(IC50)为120.6μg/mL。用浓度为0.1×IC50的CMG预处理HepG2细胞24 h,再给予2 Gy X射线或12C6+离子束辐照(CMG+辐照组);CMG未处理组直接接受2 Gy X射线或12C6+离子束辐照(辐照组)。对比分析辐照组和CMG+辐照组细胞的克隆存活、DNA损伤、凋亡与周期分布、细胞内活性氧(ROS)水平。发现:与X射线辐照组相比,相同剂量的12C6+离子辐照组克隆存活率更小,DNA损伤和周期阻滞更加严重,细胞凋亡率和细胞内ROS水平也更高。与单独X射线或12C6+离子束辐照组相比,CMG+辐照组克隆存活率明显降低,细胞凋亡率随辐照后CMG作用时间的延长而明显增加,CMG使辐照后细胞内ROS维持在一个较高的水平,同时CMG明显加重了单独辐照诱导的DNA损伤和周期阻滞。结果表明,与X射线相比,HepG2细胞对相同剂量的12C6+离子辐射更敏感;CMG可增加HepG2细胞对X射线或12C6+离子辐射的敏感性;CMG可能通过增加受照HepG2细胞内的ROS水平,加剧辐照诱导的DNA损伤,促进辐射诱导细胞凋亡而起到辐射增敏作用。This study aims to investigate the effect of carboxymethy-β-1, 3-glucan (CMG) on the sensitivity of human hepatoma HepG2 cells to X-rays or 12C6+ ions irradiation. First, the inhibitory effect of CMG on the growth of HepG2 cells was detected by CCK-8 assay, and the half maximal inhibitory concentration (IC50) was 120.6 μg/mL. HepG2 cells were pretreated with CMG at a concentration of 0.1×IC50 for 24 h and then irradiated with 2 Gy X-ray or 12C6+ ion beams (CMG + irradiation group). CMG untreated group was directly irradiated by 2 Gy X-rays or 12C6+ ions beam (irradiation group). The clone survival, DNA damage, cell apoptosis, cell cycle distribution, and intracellular reactive oxygen species (ROS) levels in irradiation group and CMG + irradiation group were comparatively analyzed. The results showed that the clone survival rate was lower, DNA damage and cycle arrest were more serious, and the rate of apoptosis and intracellular ROS levels were higher in 12C6+ ions irradiation group than those in the same dose of X-rays irradiation group. Compared with X-rays or 12C6+ ions irradiation group, the clone survival rate of CMG + irradiation group was significantly decreased, and the apoptosis rate significantly increased with the prolongation of CMG treatment post-irradiation; CMG maintained intracellular ROS at a higher level after irradiation, CMG also significantly aggravated radiation-induced DNA damage and cycle arrest. These results indicated that HepG2 cells were more sensitive to 12C6+ ions radiation than those at the same dose of X-rays. CMG increased the sensitivity of HepG2 cells to X-rays or 12C6+ ions irradiation by increasing intracellular ROS level, exacerbating radiation-induced DNA damage and promoting radiation-induced apoptosis in irradiated HepG2 cells.  相似文献   

6.
研究了和厚朴酚(HNK)对非小细胞肺癌(NSCLC)细胞系A549和H1299对低线性能量转移(LET) X射线和高LET碳离子的辐射增敏效应。首先用CCK-8检测了HNK对A549和H1299细胞的生长抑制情况,发现20 μmol/L的HNK处理对细胞的生长抑制作用较弱。用该浓度HNK预处理细胞2 h后给予不同剂量X射线或碳离子的照射,克隆存活法检测细胞的辐射敏感性,Annexin-PI双染法检测细胞凋亡,γH2AX焦点法检测DNA的双链断裂(DSB)损伤。实验结果显示:与X射线相比,NSCLC细胞对碳离子更敏感,HNK预处理仅对碳离子照射有辐射增敏作用;与碳离子单独照射相比,HNK预处理联合碳离子照射诱导了更明显的细胞凋亡;在照射后24 h,HNK预处理联合碳离子照射引起的细胞γH2AX焦点阳性率维持在较高水平,而X射线照射没有这些效应。实验结果表明,HNK预处理抑制了NSCLC细胞DNA的DSB修复,诱导了细胞凋亡的发生,从而提高了细胞对碳离子的辐射敏感性。The radiosensitizing effect of Honokiol (HNK) on non-small cell lung carcinoma (NSCLC) cell lines A549 and H1299 to low-linear energy transfer (LET) X-rays and high-LET carbon ions was investigated in this study. First, the inhibitory effects of HNK on the growth of A549 and H1299 cells were detected by CCK-8 assay, and 20 μmol/L HNK treatment was found to induce a growth inhibitory effect slightly in these two cell lines. Cells were pre-treated with HNK and then irradiated with X-rays and carbon ions of different doses. Cellular radiosensitivity, apoptosis and DNA damage were analyzed by clonogenic survival, Annexin-PI staining and γH2AX foci, respectively. The results showed the cells were more sensitive to carbon ion irradiation compared to X-rays and the radiosensitization of HNK was only observed after carbon ion irradiation. Furthermore, the co-treatment led to higher apoptosis rate 48 h after irradiation and increased the positive rate of γH2AX foci 24 h after irradiation in A549 and H1299 cells compared with those in the groups treated with carbon ion irradiation alone. These phenomena were not observed after X-ray irradiation. Our data suggest that the pre-treatment with HNK inhibited DNA DSB repair, induced apoptosis and then enhanced the cellular radiosensitivity to carbon ions in NSCLC cells.  相似文献   

7.
辐照诱导的人正常肝细胞系HL-7702细胞延迟效应   总被引:1,自引:0,他引:1  
利用X射线辐照人正常肝细胞系HL-7702细胞, 运用胞质分离阻滞微核法实验检测细胞微核率,AnnexinV FITC细胞凋亡检测试剂盒检测细胞凋亡率, 细胞微核率和凋亡率随着辐照剂量的增加而显著增加。X射线照射后细胞传代培养, 第7代时不同剂量辐照后子代细胞微核率和凋亡率同未辐照细胞相比已无明显区别。 对不同剂量辐照后传代7代的细胞再次照射2.5 Gy的相同剂量,发现它们细胞微核率和凋亡率存在明显差异,即初次受辐照剂量高的细胞, 再次以相同剂量辐照后的微核率和凋亡率也高. 这些结果表明,X射线辐照导致了HL-7702细胞基因组不稳定性这一辐射延迟效应,再次辐照使得辐射的延迟效应得以明显的表现。 Human normal liver cell line HL-7702 cells were irradiated with different doses of X rays. Micronucleus and apoptosis rates in the irradiated cells were measured with cytokinesis block micronucleus method and Annexin V FITC apoptosis detection kit, respectively. Experimental data showed that the micronucleus and apoptosis rates increased obviously with increasing irradiation dose. After seven population doublings, the micronucleus and apoptosis rates of the cells surviving exposure to the X rays reduced to the same levels as non irradiated control cells; the progenies of the cells were secondly exposed to X rays at the same dose of 2.5 Gy. We found that the progenies of the cells surviving the first irradiations of the various doses showed markedly differential micronucleus frequencies and apoptotic rates. Although the same dose of 2.5 Gy was applied in the second irradiations, the micronucleus frequencies and apoptotic rates of the progenies of the cells initially exposed at higher doses were significantly higher than the others. These results indicate that X rays lead to genomic instability in HL 7702 cells, which is an important manifestation of radiation induced delayed effect, and a second radiation stimulus makes the delayed effect in the progeny of the previously irradiated cells be expressed obviously.  相似文献   

8.
以HeLa细胞为实验材料, 探讨了NADPH氧化酶在X射线诱导细胞损伤过程中的作用。 结果显示, 12 Gy X射线辐照后细胞内活性氧(ROS)明显增加, 在用NADPH氧化酶抑制剂处理后再辐照, 则细胞内ROS降低到未辐照水平; 同时辐照后NADPH 氧化酶细胞质亚基p47phox 在细胞质积聚并和细胞膜亚基 gp91phox 结合; Western blotting检测结果显示, NADPH 氧化酶的关键亚基 gp91phox 的表达量明显增加。 以上结果说明, NADPH氧化酶可以被X射线激活, 由其介导产生的ROS在X射线诱导HeLa细胞损伤过程中扮演重要角色。 To investigate the role of NADPH oxidase in HeLa cell lesion induced by X ray irradiation, the change of cell survival was detected with MTT assay, reactive oxygen species (ROS) was measured by fluorospectrophotometer. Immunostaining and confocal laser scanning microscopy was employed to detect the co localization of two subunit of NADPH oxidase, p47phox and gp91phox in the cell. Western blotting was used to detect the expression of gp91phox before and after X ray irradiation. After X ray irradiation, intra cellular level of ROS increased obviously. But the increase could be blocked by diphenyleneiodonium (DPI), an inhibitor of NADPH oxidase. Meanwhile, cytosolic subunit p47phox moved to membrane and co localizated with gp91phox after irradiation. Moreover, the results also show that gp91phox increased sharply after 12 Gy X ray irradiation. Therefore, NADPH oxidase mediated production of ROS plays an important role in HeLa cell lesion induced by X ray.  相似文献   

9.
We investigate the temperature dependence of radiation-induced attenuation (RIA) at 1 310 nm for a Ge/P co-doped fiber after a steady-state γ-ray irradiation.A γ irradiation facility 60Co source is used to irradiate the fiber at a dose rate of 0.5 Gy/min,satisfying a total dose of 100 Gy.The test temperature ranges from-40 to 60℃ by 20℃,and the RIA of the fiber is obtained using a power measuring device.The experimental result demonstrates that RIA exhibits a steady,monotonic,and remarkable temperature dependence after approximately 48h of accelerated annealing at 70℃.The optical fiber irradiated with a high dose and annealed sufficiently can be used as a temperature sensor.  相似文献   

10.
综述了DNA辐射损伤导致的细胞阻遏于G1期以及在该时期对DNA的修复活动, 提出了较大剂量辐射诱导的三磷酸腺苷不足导致细胞凋亡的假说, 并分析了细胞走向凋亡与修复的辨正关系。 DNA damage induced by irradiation,which makes the cell arrested at G1 stage and DNA repair being activated in this stage,are summarized. It is proposed that the deficiency of adenosine triphosphate which is induced by the larger irradiation dose, induces cell apoptosis. And the relationship of cell selecting repair and apoptosis is also analyzed.  相似文献   

11.
以0, 0.05, 0.1, 0.25, 0.5 Gy 12C6+ 离子全身预辐照昆明小鼠, 间隔4 h后再对小鼠进行4 Gy全身辐射。 辐照后12 h用流式细胞仪检测小鼠胸腺脾脏细胞在各细胞周期时相的百分率, 同时用单细胞电泳检测受辐射小鼠胸腺脾脏细胞DNA损伤程度。 结果显示, 相对于大剂量预照射组, 各低剂量预照射组胸腺细胞S期细胞百分率显著减少; 脾脏细胞G0/G1期细胞百分率明显减少; 同时胸腺脾脏细胞的拖尾率及拖尾长度明显减少, 以0.1 Gy预辐照效果最为明显。 这些结果表明, 低剂量预辐射处理可以减弱胸腺细胞的S期阻滞及脾脏细胞的G1期阻滞, 并明显减轻胸腺脾脏细胞的DNA损伤程度。  相似文献   

12.
Fourier transform infrared spectroscopy (FTIR) was employed to study the human epidermis larynx carcinoma cell lines (Hep-2) which were irradiated by different doses of X-ray.The results show that (1) the irradiation of X-ray damages the structure of the CH3 groups of the thymine in DNA,which restrains the reproduction of Hep-2 cells effectively,(2) the 8 Gy dose of X-ray irradiation changes the framework and the relative contents of some proteins,lipids and the nucleic acid molecules intercellular in the greatest degree,and (3) the 8 Gy dose of X-ray irradiation is the best irradiation dose for lowering the degree of the cancerization of Hep-2 cells according to the criteria for the degree of the cancerization reported recently.Meanwhile,the apoptosis of these cells were detected by using flow cytometry (FCM) primarily.It shows that the apoptotic ratio of the Hep-2 cells depends on the irradiation dose to some extent,but is not linearly.And the apoptotic ratio of the 12 Gy dose group is the maximum (20.36%),but the apoptotic ratios of the 2 to 8 Gy dose groups change little.  相似文献   

13.
采用高传能线密度(LET)重离子辐照人胃癌SGC7901细胞,应用流式细胞技术、蛋白质印迹法(Western blot)及反转录聚合酶链式反应(RT-PCR)观察重离子诱导人胃癌SGC7901细胞周期、凋亡和MSH2表达状况。结果表明:与对照组相比,SGC7901细胞在辐射后72 h G2/M期所占细胞比率(33.26±0.08)和凋亡率(24.16±0.64)均达到峰值,且呈时间依赖性增加;经重离子照射后,DNA错配修复基因MSH2 m RNA和蛋白表达水平在6 h最高。结果提示:重离子在体外诱导SGC7901细胞周期阻滞和凋亡,且具有显著的时间依赖性效应;重离子在一定剂量和时间下,诱导了SGC7901细胞MSH2基因表达。DNA错配修复基因MSH2可能参与了重离子辐照诱导胃癌细胞DNA损伤的修复应答。  相似文献   

14.
低剂量电离辐射引发的生物效应复杂而多样,其研究往往又受到辐射标志物和检测技术手段的限制。将拉曼光谱技术应用于低剂量辐射生物效应研究,利用10 mW,532 nm共聚焦拉曼光谱对经过100,200和500 mGy三种辐射剂量的X射线辐照之后的人神经母细胞瘤细胞进行检测,发现细胞嘌呤核苷酸(722~728和1 572~1 581 cm-1等等)、嘧啶核苷酸(770~785 cm-1等等)等DNA相关的拉曼特征峰受到电离辐射影响而发生变化,说明低剂量X射线辐照造成细胞DNA水平改变。采用流式细胞术对同样条件辐照后培养6 h的人神经母细胞瘤细胞进行细胞周期分析发现,三种剂量的X射线电离辐射均造成细胞在G2期阻滞,同样提示电离辐射引起DNA水平升高。通过划痕实验分析辐照后20 h的细胞迁移能力,结果显示,相较于未接受X射线照射的对照细胞,受到三种剂量电离辐射的人神经母细胞瘤细胞均出现迁移水平下降。研究结果表明,通过拉曼光谱分析发现低剂量X射线电离辐射引起人神经母细胞瘤细胞DNA水平变化,其结果与细胞周期分析和迁移分析的结果相一致,但检测时间大大提前,利用拉曼光谱技术可以实现低剂量辐射损伤等细胞生物学效应的早期发现与监测。  相似文献   

15.
The effect of lower doses (0.5–3.0 Gy) of gamma radiation on radiosensitivity of CD3?/CD8+ NK cells subpopulation isolated from the peripheral blood of healthy volunteers was studied 48 h after the irradiation. Only a subtle increase in terms of induction of apoptosis (A+ cells), was observed in Annexin positive CD3?/CD8+ NK cells. The assessment of the relative presence of CD3?/CD8+ NK cells in Annexin negative populations of lymphocytes considerably contributes to the elimination of individual variability and could be useful in biodosimetry.Living CD3?/CD8+; Annexin negative NK cells were analyzed using five-color flow cytometry 16 h after irradiation by the doses of 1–10 Gy. The study was carried out on NK cells subsets CD3?/CD8? CD16+, CD56 (dim) and CD56 (bright). NK cells characterized with their low-density expression of CD56 (dim) are more cytotoxic and express CD16. Those with high-density expression of CD56 (bright) are known for their capacity to produce cytokines following activation of monocytes but their natural cytotoxicity is low; they are classified as CD16? or CD16 (dim). A dose-depending decrease in the relative presence of CD3?/CD8+ NK cells was observed 16 h after ionizing radiation (1–10 Gy). The decrease was highly pronounced in CD56 (bright) subset of NK cells and this subpopulation was considered as the most radiosensitive one. Unfortunately, the most radiosensitive subpopulation of NK cells – CD56bright cannot be used as a biodosimetric marker due to its insufficient amount in peripheral blood.  相似文献   

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