RNA干扰沉默PDGFR-β基因增敏放射治疗C6脑胶质瘤移植瘤的研究

发布时间：2018-03-03 20:10

本文选题：RNA干扰　切入点：血小板源生长因子受体-β　出处：《中南大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：本课题前期实验已证实RNA干扰PDGFR-β基因能抑制C6胶质瘤细胞PDGFR-β的表达,并提高C6胶质瘤细胞体外放射敏感性。本实验探索RNA干扰沉默PDGFR-β表达联合放射治疗对胶质瘤裸鼠移植瘤生长的影响。方法：将50只BALB/c裸鼠随机分为5组(n=10)：对照组、RNAi-PDGFR-β组、空载病毒组、放射治疗组、联合治疗组。用不同C6细胞悬液分别接种在4-6周的雌性裸鼠左前肢的腋后方皮下建立裸鼠移植瘤模型。待皮下瘤体积增至100-200mmm3时,对放射治疗组和联合治疗组行放射治疗,治疗后48小时,每组处死4只裸鼠,取移植瘤行免疫组化检测PDGFR-β、Ki-67、Cyclin B1、VEGF表达及细胞凋亡检测。其余6只裸鼠继续观察肿瘤的生长情况,每3天测量一次移植瘤的最长直径(L)和最短直径(W),利用公式V(mm3)=1/2(L×W2)(mm3)计算肿瘤体积,根据肿瘤的体积绘制不同组别裸鼠移植瘤的生长曲线。28天时断颈处死所有的裸鼠,分离取出皮下肿瘤,称重并计算抑瘤率。结果：空载病毒组、RNAi-PDGFR-β组、放射治疗组和联合治疗组的瘤重抑瘤率分别为0.27%±9.9%,0.05%±8.65%,57.58%±6.74%,73.26%±3.65%。放射治疗组、联合治疗组与其他三组比较,移植瘤的体积明显减小,联合治疗组最小,对照组、空载病毒组、RNAi-PDGFR-β组的肿瘤体积差别无显著性,联合治疗组与放疗组的抑瘤率与对照组,空载病毒组和RNAi-PDGFR-β组的差异有统计学意义((P0.05))。放射治疗和联合治疗组与其他三组相比,能明显的抑制C6胶质瘤细胞PDGFR-β、Ki-67、Cyclin B1的表达,增加细胞凋亡,而联合治疗组比放疗组的作用更显著。联合治疗组与对照组、空载病毒组、RNAi-PDGFR-β组相比,能减低VEGF的表达,而放射治疗组能使VEGF的表达增多。结论：RNA干扰抑制PDGFR-β表达联合放疗,使胶质瘤细胞增殖抑制,侵袭性减弱、凋亡增加,较单纯的放疗具有更好的抗肿瘤细胞作用。RNA干扰抑制PDGFR-β表达能增加胶质瘤放射敏感性,PDGFR-β可作为胶质母细胞瘤治疗的靶点之一。
[Abstract]:Objective: this study has demonstrated that RNA interference of PDGFR- 尾 gene can inhibit the expression of PDGFR- 尾 in C6 glioma cells. In order to improve the radiosensitivity of C6 glioma cells in vitro, we investigated the effects of RNA interference silencing PDGFR- 尾 expression combined with radiotherapy on the growth of glioma xenografts in nude mice. Methods: fifty BALB/c nude mice were randomly divided into 5 groups: control group (n = 5): RNAi-PDGFR- 尾 group, no-load virus group and radiotherapy group. In the combined treatment group, the transplanted tumor model of nude mice was established by inoculating different C6 cell suspensions at the posterior axillary of the left forelimb of female nude mice for 4-6 weeks. When the volume of subcutaneous tumor increased to 100-200 mm ~ 3, the radiotherapy group and the combined treatment group were treated with radiotherapy. 48 hours after treatment, 4 nude mice were killed in each group. The expression of PDGFR- 尾 Ki-67-Cyclin B1C VEGF and apoptosis were detected by immunohistochemistry. The other 6 nude mice continued to observe the growth of tumor. The longest diameter L) and the shortest diameter of transplanted tumor were measured every 3 days. The tumor volume was calculated by using the formula V ~ (mm ~ (3) / L 脳 W ~ (2 +)). The growth curve of different groups of transplanted tumor was drawn according to the volume of tumor. At 28 days, all the nude mice were killed by cervical breakage. The subcutaneous tumor was removed and weighed and the tumor inhibition rate was calculated. Results: the tumor weight inhibition rates in the non-loaded virus group, radiotherapy group and combined treatment group were 0.27% 卤9.9g% 卤8.65% 卤8.65% 卤7.58% 卤6.74% 卤3.65% respectively. Compared with the other three groups, the volume of transplanted tumor in radiotherapy group, combined treatment group and combined treatment group was significantly decreased, while that in combined treatment group and control group was the smallest. There was no significant difference in tumor volume in RNAi-PDGFR- 尾 group. The tumor inhibition rate of combined treatment group and radiotherapy group was significantly higher than that of control group, empty virus group and RNAi-PDGFR- 尾 group. The expression of PDGFR- 尾 -ki-67-cyclin B1 in C6 glioma cells was significantly inhibited, and the apoptosis was increased in the combined treatment group than in the radiotherapy group. The expression of VEGF in the combined treatment group was significantly lower than that in the control group and the no-load virus group, and the expression of VEGF was decreased in the combined treatment group compared with the control group and the no-load virus group. In radiotherapy group, the expression of VEGF was increased. Conclusion the inhibition of the expression of PDGFR- 尾 by the interference of fraction RNA combined with radiotherapy can inhibit the proliferation, decrease the invasion and increase the apoptosis of glioma cells. The inhibition of PDGFR- 尾 expression by RNA interference could increase the radiosensitivity of gliomas. PDGFR- 尾 could be used as a target for glioblastoma treatment.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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