低剂量辐射通过调节Survivin和Caspase-3的表达水平增强卵巢癌耐药细胞的顺铂敏感性

发布时间：2018-03-29 18:27

本文选题：SKOV3/DDP　切入点：低剂量辐射　出处：《青岛大学》2016年硕士论文

【摘要】：目的:顺铂是卵巢癌的主要化疗药物,然而在临床应用中卵巢癌细胞往往会呈现出对顺铂的耐药现象。前期研究发现肿瘤细胞对低剂量辐射敏感,低剂量辐射治疗可以提高卵巢癌对化疗药物的敏感性,但是机制却不是很清楚。本课题拟将研究低剂量辐射对卵巢癌顺铂耐药细胞的影响,并从Survivin和Caspase-3基因水平,了解低剂量辐射对卵巢癌顺铂耐药细胞的影响机制。方法:卵巢癌顺铂耐药(SKOV3/DDP)细胞随机分为三组,常规照射组、低剂量辐射组(LDR)、对照组,每组的顺铂浓度梯度均为0,0.625,1.25,2.5,5和10ug/L。于0,8,16,24时给予低剂量辐射组0.5Gy照射,照射总量为2Gy。常规组给予2Gy一次的照射,对照组不给予照射。CCK8用于检测低剂量辐射对SKOV3/DDP细胞活性的影响,流式细胞术用于检测肿瘤细胞的凋亡,real-time PCR技术检测肿瘤细胞耐药相关蛋白Caspase-3和Survivin的表达。结果:与对照组和常规组相比,低剂量辐射组的细胞增殖活性明显降低(P0.05)。低剂量照射组,对照组和常规照射组的IC50分别是:3.837±0.16,9.467±0.17和9.389±0.17。低剂量辐射组的凋亡率明显升高,顺铂浓度为2.5ug/L时,低剂量辐射组的凋亡率为51.40%±0.24,对照组为20.70%±0.59,常规剂量组为39.20%±0.29。低剂量辐射组的Caspase-3表达水平明显高于常规剂量组、对照组,而Survivin表达水平低于对照组和常规照射组(P0.05)。结论:在卵巢癌组织中Survivin的表达水平高于正常卵巢组织,Caspase-3的表达水平低于正常卵巢组织。Survivin和Caspase-3的表达对卵巢癌的形成有重要的作用。本实验研究发现卵巢癌耐药细胞经低剂量照射后,细胞中的Survivin的表达水平降低,Caspase-3的表达水平升高。我们得出结论,低剂量辐射或许能够通过调节Survivin和Caspase-3的表达来增强卵巢癌细胞对顺铂的敏感性,提高卵巢癌的临床治疗效果,为卵巢癌的临床治疗提供新的方法。
[Abstract]:Objective: cisplatin is the main chemotherapeutic agent for ovarian cancer. However, in clinical application, ovarian cancer cells tend to be resistant to cisplatin. Previous studies have found that tumor cells are sensitive to low dose radiation. Low dose radiation therapy can improve the sensitivity of ovarian cancer to chemotherapeutic drugs, but the mechanism is not very clear. In this study, we will study the effects of low dose radiation on cisplatin resistant ovarian cancer cells, and study the gene levels of Survivin and Caspase-3. To understand the mechanism of low dose radiation on cisplatin resistant ovarian cancer cells methods: ovarian cancer Cisplatin resistant SKOV3 / DDP cells were randomly divided into three groups: conventional radiation group, low dose radiation group, control group. The gradient of cisplatin concentration in each group was 0 ~ 0.625 ~ 1.25 ~ 2.5g / L and 10 g / L 路L ~ (-1). The low dose radiation group was given 0.5Gy irradiation at 24:00 and the total dose was 2Gy.The routine group was given 2Gy once, while the control group was not given the irradiation. CCK8 was used to detect the effect of low dose radiation on the activity of SKOV3/DDP cells. Flow cytometry was used to detect the expression of Caspase-3 and Survivin in tumor cells by real-time PCR. The cell proliferation activity of low dose radiation group was significantly lower than that of low dose radiation group. The IC50 of low dose radiation group, control group and conventional irradiation group were respectively 3. 837 卤0. 169.467 卤0. 17 and 9.389 卤0. 17.The apoptotic rate of low dose radiation group was significantly higher than that of normal irradiation group, when the concentration of cisplatin was 2.5ug/L. The apoptotic rate was 51.40% 卤0.24 in low dose radiation group, 20.70% 卤0.59 in control group and 39.20% 卤0.29 in conventional dose group. The expression of Caspase-3 in low dose radiation group was significantly higher than that in normal dose group and control group. Conclusion: the expression level of Survivin in ovarian cancer is higher than that in normal ovarian tissue. The expression of survivin and Caspase-3 in ovarian cancer is lower than that in normal ovarian tissue. This study found that ovarian cancer drug-resistant cells were exposed to low doses of radiation, We conclude that low dose radiation may enhance the sensitivity of ovarian cancer cells to cisplatin by regulating the expression of Survivin and Caspase-3, and improve the clinical therapeutic effect of ovarian cancer. To provide a new method for the clinical treatment of ovarian cancer.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R737.31

【参考文献】