【作者】 彭阳红；

【导师】 周乃康；

【作者基本信息】 中国人民解放军军医进修学院 ， 外科学， 2011， 博士

【摘要】 目的：1、构建125I-103Pd复合性放射性粒子,并与125I和103Pd粒子比较,评价其对肿瘤细胞杀伤效应及抑瘤效果；2、比较125I-103Pd与125I及103Pd三种粒子对肿瘤细胞凋亡、增殖及代谢的影响,探索125I₁03Pd复合粒子对肿瘤细胞杀伤的机理。方法：1、采用化学镀层的方法构建125I₁03Pd复合性放射性粒子,采用自显影及粒子活度测定评价粒子的均一性；2、细胞学实验：选取A549、GLC-82、95D及H460四株肺癌细胞,进行细胞杀伤、凋亡及增殖实验,比较125I-103Pd与125I及103Pd三种粒子对肿瘤细胞体外生长的影响。3、小鼠移植瘤粒子植入治疗实验：建立小鼠移植瘤模型,比较125I-103Pd与125I及103Pd三种粒子的抑瘤效果；4、PET-CT分子影像学实验：采用18F-FDG、18F-FLT及11C-CH三种显像剂进行小鼠肿瘤A549及95D移植瘤模型显像,比较125I-103Pd与125I及103Pd三种粒子对肿瘤代谢及增殖的影响,探索125I-103Pd复合粒子对肿瘤细胞杀伤的机理。结果：1、粒子组自显影及活度测定表明,125I-103Pd复合粒子构建成功,均一性可靠；2、体外粒子对肿瘤细胞杀伤实验表明,48h内103Pd粒子效果最好,125I-103Pd复合粒子次之,125I杀伤效果最弱,细胞杀伤有效范围未见明显差异,粒子周围2-3个粒子直径范围内四种肿瘤细胞的生长均受到明显抑制。3、细胞凋亡实验表明,2Gy照射剂量下,125I-103Pd与125I及103Pd粒子对A549细胞凋亡的影响无明显差异,对于95D细胞,103Pd粒子诱导细胞凋亡及死亡的作用较125I-103Pd及125I粒子强,125I-103Pd与125I粒子间差异不明显；4Gy照射剂量下,对于A549及95D细胞,125I-103Pd复合粒子促凋亡作用较125I及103Pd粒子强,而103Pd粒子对肿瘤细胞的致死效应较125I-103Pd及125I粒子明显。4、体外细胞增殖实验表明,103Pd粒子在第1～4天,对肿瘤细胞生长的抑制效果强于125I-103Pd及125I粒子（P<0.05）,第5～7天125I-103Pd与103Pd粒子对肿瘤细胞生长的抑制效果相当（P>0.05）；125I粒子对肿瘤增殖的影响最小（P<0.05）；5、体内移植瘤粒子植入治疗实验表明,125I-103Pd复合粒子抑瘤作用强于103Pd和125I粒子（P<0.05）,103Pd和125I粒子抑瘤效果无明显差异；6、A549移植瘤模型的PET-CT结果表明：125I-103Pd、125I及103Pd粒子组的葡萄糖及核苷酸代谢明显低于空白粒子组；103Pd粒子对肿瘤的葡萄糖代谢影响最明显,125I-103Pd复合粒子次之,而125I粒子最弱；粒子植入初期,103Pd粒子对肿瘤的核苷酸代谢影响最明显,而中后期125I-103Pd复合粒子对核苷酸代谢影响最明显；125I粒子在治疗初期对肿瘤核苷酸代谢影响最弱,中后期与103Pd粒子作用相当；肿瘤A549核苷酸代谢与葡萄糖代谢的趋势一致,随着观察时间的延长,摄取值均降低。7、95D移植瘤模型的PET-CT结果表明：125I-103Pd复合粒子对肿瘤95D的葡萄糖及核苷酸代谢的影响较125I粒子大；95D肿瘤核苷酸代谢与葡萄糖代谢趋势不一致：糖代谢水平持续走低,而核苷酸代谢在治疗中后期无明显差异。8、125I-103Pd,125I及103Pd粒子对肿瘤的磷脂代谢影响明显,但三个粒子组间比较无明显差异。结论：1、125I-103Pd复合性放射性粒子制备工艺可靠,粒子均一性较好；2、125I-103Pd与125I及103Pd粒子比较,其有效杀伤范围未见明显差异；3、125I粒子主要是通过诱导凋亡来杀伤肿瘤细胞；103Pd粒子则主要通过射线的直接杀伤作用,引起细胞死亡；125I-103Pd复合粒子则兼顾了两种核素的特性；4、125I-103Pd复合粒子的抑瘤效果强于103Pd与125I粒子,103Pd与125I粒子的抑瘤效果相当；5、125I-103Pd与125I及103Pd粒子对肿瘤的葡萄糖代谢、核苷酸代谢及磷脂代谢影响均较明显,但125I-103Pd复合粒子对肿瘤核苷酸代谢的影响较125I及103Pd粒子明显；核苷酸代谢水平高低更能反映粒子治疗后的抑瘤效果。更多还原

【Abstract】 Objective:1. To develop 125I- 103Pd hybrid radioactive seeds, and to observe the killing effects and the tumor-inhibitory action of 125I- 103Pd hybrid radioactive seeds, by comparing with 125I and 103Pd radioactive seeds.2. To investigate the killing mechanism of 125I-103Pd seeds, by comparing with 125I and 103Pd radioactive seeds on tumor cells apoptosis, proliferation and metabolism.Method:1.125I- 103Pd hybrid radioactive seeds was developed by using chemical plating method, and the homogenicity of the seed was estimated by nuclide autograph and the measurement of nuclide seed activity.2. Cytological test:4 kind of lung cancer cells including A549, GLC-82,95D and H460 cells were chosen for tumor cell killing test, cell apoptosis test and cell proliferation test, in order to compare the effects of 125I- 103Pd seeds with 125I and 103Pd seeds on tumor cell growth in vitro.3. Seed implanting therapeutic test:the mouse models of transplanted tumors were built and the tumor-inhibitory action was compared between 125I- 103Pd,125I and 103Pd seeds.4. Molecular imaging test:tracer agents including 18F-FDG、18F-FLT and 11C-CH were adopted for PET examination on A549 and 95D tumor models, to learn about the effects of 125I- 103Pd,125I and 103Pd seeds on tumor cell metabolism and proliferation, in order to investigate the the killing mechanism of 125I- 103Pd seed on tumor cells.Result:1. Nuclide autograph and the measurement of nuclide seed activity indicated that, 125I- 103Pd seed was successfully developed, and thehomogenicity of the seed was reliable;2. In vitro cell killing test in 48 hours showed that,103Pd seed was the most effective, while 125I- 103Pd seed was more effective than 125I seed. The killing ranges of the three kind of radioactive seeds were of no significant difference, and tumor cells around radioactive seeds in a range of about 2 to 3 lengths of a seed were inhibited obviously。3. Cell apoptosis test indicated that, under 2Gy irradiation, no obvious difference in A549 cell apoptosis was observed among 125I- 103Pd,125I and 103Pd seeds (P>0.05). While under 2Gy irradiation,103Pd seed could induce more 95D cells to apoptosis and death than 125I-103Pd and 125I seeds (P<0.05), no difference was found between 25I- 103Pd and 125I seeds (P>0.05). Under 4Gy irradiation, as regard to both A549 and 95D cells, 125I- 103Pd seed was more effective in inducing tumor cell to apoptosis than 125I and 103Pd seeds did, while 103Pd seed could induce more tumor cells to death than 125I-103Pd and 125I seeds did.4. Tumor cell proliferation test in vitro showed that:From day 1 to day 4,103Pd seed was much more effective in cell growth inhibition than 125I- 103Pd and 125I seeds(P<0.05); From day 5 to day 7,125I- 103Pd and 103Pd seeds were identical in cell growth inhibition (P>0.05). While 125I seed was relatively the weakest radioactive seed in whole observation period (P<0.05);5. Seed implanting test in transplanted tumors showed that, 125I- 103Pd seed was the most effective in tumor inhibition, while 103Pd and 125I seeds were identical;6. PET-CT of A549 tumor models indicated that,125I- 103Pd,125I and 103Pd seeds could obviously inhibit A549 cells metabolism in glucose and nucleotide. As to glucose metabolism,103Pd seed is the more powerful than 125I-103Pd and 125I seeds, while 125I seeds is the weakest among the three kind of radioactive seeds. In the early observation time,103Pd seed was the most effective in the inhibition of the nucleotide metabolism, while 125I- 103Pd seed was the most powerful in the mid-and-later observation time; 125I seed was the weakest in the early observation time, while it was as effective as 103Pd seed in the mid-and-later observation time. The tendency of glucose metabolism is concordant to nucleotide metabolism, while the uptake value descended gradually along with the time of irradiation.7. PET-CT of 95D tumor models indicated that,125I- 103Pd was more effective than 125I seeds to inhibit cell metabolism in glucose and nucleotide. However, the tendency of glucose metabolism is inconcordant to nucleotide metabolism:the uptake value of glucose metabolismdescended gradually, while nucleotide metabolism was of little change in mid to late time of irradiation。8. The influence of 125I- 103Pd,125I and 103Pd seeds on the phospholipid metabolism of tumor cell were obvious. However, no difference was found among the 3 radioactive seed groups.Conclusion:1. The manufacture of 125I- 103Pd hybrid radioactive seed is possible, the homogenicity of the seed is reliable;2.125I- 103Pd,125I and 103Pd seeds are of no difference in effective killing range.3.125I seeds mainly induce tumor cells to apoptosis, and 10 Pd seeds mainly kill tumor cells directly, while 125I- 103Pd seeds may combine the character of the two nuclide.4. In tumor-inhibitory action,125I- 103Pd seed is more effective than 103Pd and 125I seeds, while 103Pd seed is comparable with 125I seed.5.125I- 103Pd,125I and 103Pd seeds can influence tumor cell metabolism in glucose, nucleotide and phospholipid. However, 125I- 103Pd seed is more powerful in inhibiting the nucleotide metabolism, than 125I and 103Pd seeds; Nucleotide metabolism could better reflect the tumor-inhibitory effect of radioactive seeds.更多还原