硼替佐米通过抑制HIF-1α的表达对乏氧宫颈癌细胞株产生放疗增敏作用的研究

发布时间：2018-01-14 15:01

本文关键词：硼替佐米通过抑制HIF-1α的表达对乏氧宫颈癌细胞株产生放疗增敏作用的研究　出处：《蚌埠医学院》2015年硕士论文　论文类型：学位论文

【摘要】：本研究旨在探讨硼替佐米对于宫颈癌的放疗增敏作用及可能的机制。宫颈癌是最常见的的妇科恶性肿瘤之一,放射治疗是其主要治疗方法,目前仍有相当比例的患者存在放射治疗抵抗。实体瘤中乏氧细胞约占10%-50%,对放射线有较强的抵抗力。有文献报道女性宫颈癌中肿瘤氧合好的患者放疗后无复发生存率明显高于氧合差的患者。硼替佐米能够特异性抑制哺乳动物细胞内26S蛋白酶体的类凝乳胰蛋白酶活性,通过降解调节蛋白,产生抗增殖、促凋亡、抗血管生成,从而起到抑制肿瘤活性的作用。分别把常氧和乏氧两种状态下的宫颈癌细胞系HeLa细胞和SiHa细胞分为对照组、放射治疗组、药物组(低浓度、高浓度)和放射治疗联合药物组。通过CCK8实验检测细胞增殖能力,进而选择药物浓度；通过克隆形成实验比较集落形成数量,得到常氧状态和乏氧状态下硼替佐米对于两种细胞的放射增敏比(SER)；随后通过流式细胞术检测细胞凋亡；免疫荧光实验检测DNA双链断裂(DSB); Western blot实验检测HIF-1α、PARP-1和Caspase-3、8、9的表达情况。数据统计分析表明,纳摩尔水平的硼替佐米对常氧和乏氧两种状态下的宫颈癌细胞均具有放射治疗增敏作用。Western blot实验表明该药物能抑制HIF-la的表达、促进与凋亡相关的Caspase-3、8、9的激活、增加与DBS相关的PARP-1裂解。因此,硼替佐米可能是潜在的宫颈癌放射治疗增敏剂。
[Abstract]:The purpose of this study was to investigate the radiosensitizing effect of bortezomil on cervical cancer and its possible mechanism. Cervical cancer is one of the most common gynecological malignancies and radiotherapy is the main treatment. At present, there is still a considerable proportion of patients with radiotherapy resistance. The proportion of hypoxic cells in solid tumors is about 10-50%. It has been reported that the recurrence free survival rate of female cervical cancer patients with good oxygenation after radiotherapy is significantly higher than that of patients with poor oxygenation. Bortezomil can specifically inhibit mammalian cell 2. The chymotrypsin activity of 6s proteasome. Through the degradation of regulatory proteins, the production of anti-proliferation, apoptosis, anti-angiogenesis. The cervical cancer cell line HeLa and SiHa were divided into control group, radiotherapy group and drug group (low concentration). CCK8 assay was used to test the cell proliferation ability, and then the drug concentration was selected. By comparing the number of colony formation with clone formation experiments, the radiosensitization ratio of Bortezomil to the two kinds of cells was obtained in normoxic and hypoxic state. Apoptosis was detected by flow cytometry. The double strand breaks of DNA were detected by immunofluorescence assay. Western blot assay was used to detect the expression of HIF-1 伪 -PARP-1 and Caspase-3 and Caspase-3, and the statistical analysis showed that the expression of PARP-1 and Caspase-3 was detected by statistical analysis. Bortezomil at nanomolor level can sensitize cervical cancer cells in both normoxic and hypoxic states by radiotherapy. Western. Blot assay showed that the drug could inhibit the expression of HIF-la. The activation of apoptosis-related Caspase-3 and caspase-8, and the increase of PARP-1 cleavage associated with DBS were promoted. Therefore, bortezomil may be a potential radiosensitizer for cervical cancer.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.33

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