本文選題：RNA干擾　切入點(diǎn)：血小板源生長因子受體-β　出處：《中南大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：本課題前期實(shí)驗(yàn)已證實(shí)RNA干擾PDGFR-β基因能抑制C6膠質(zhì)瘤細(xì)胞PDGFR-β的表達(dá),并提高C6膠質(zhì)瘤細(xì)胞體外放射敏感性。本實(shí)驗(yàn)探索RNA干擾沉默PDGFR-β表達(dá)聯(lián)合放射治療對膠質(zhì)瘤裸鼠移植瘤生長的影響。 方法：將50只BALB/c裸鼠隨機(jī)分為5組(n=10)：對照組、RNAi-PDGFR-β組、空載病毒組、放射治療組、聯(lián)合治療組。用不同C6細(xì)胞懸液分別接種在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。待皮下瘤體積增至100-200mmm3時(shí),對放射治療組和聯(lián)合治療組行放射治療,治療后48小時(shí),每組處死4只裸鼠,取移植瘤行免疫組化檢測PDGFR-β、Ki-67、Cyclin B1、VEGF表達(dá)及細(xì)胞凋亡檢測。其余6只裸鼠繼續(xù)觀察腫瘤的生長情況,每3天測量一次移植瘤的最長直徑(L)和最短直徑(W),利用公式V(mm3)=1/2(L×W2)(mm3)計(jì)算腫瘤體積,根據(jù)腫瘤的體積繪制不同組別裸鼠移植瘤的生長曲線。28天時(shí)斷頸處死所有的裸鼠,分離取出皮下腫瘤,稱重并計(jì)算抑瘤率。 結(jié)果：空載病毒組、RNAi-PDGFR-β組、放射治療組和聯(lián)合治療組的瘤重抑瘤率分別為0.27%±9.9%,0.05%±8.65%,57.58%±6.74%,73.26%±3.65%。放射治療組、聯(lián)合治療組與其他三組比較,移植瘤的體積明顯減小,聯(lián)合治療組最小,對照組、空載病毒組、RNAi-PDGFR-β組的腫瘤體積差別無顯著性,聯(lián)合治療組與放療組的抑瘤率與對照組,空載病毒組和RNAi-PDGFR-β組的差異有統(tǒng)計(jì)學(xué)意義((P0.05))。放射治療和聯(lián)合治療組與其他三組相比,能明顯的抑制C6膠質(zhì)瘤細(xì)胞PDGFR-β、Ki-67、Cyclin B1的表達(dá),增加細(xì)胞凋亡,而聯(lián)合治療組比放療組的作用更顯著。聯(lián)合治療組與對照組、空載病毒組、RNAi-PDGFR-β組相比,能減低VEGF的表達(dá),而放射治療組能使VEGF的表達(dá)增多。 結(jié)論：RNA干擾抑制PDGFR-β表達(dá)聯(lián)合放療,使膠質(zhì)瘤細(xì)胞增殖抑制,侵襲性減弱、凋亡增加,較單純的放療具有更好的抗腫瘤細(xì)胞作用。RNA干擾抑制PDGFR-β表達(dá)能增加膠質(zhì)瘤放射敏感性,PDGFR-β可作為膠質(zhì)母細(xì)胞瘤治療的靶點(diǎn)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% 鹵9.9g% 鹵8.65% 鹵8.65% 鹵7.58% 鹵6.74% 鹵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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R739.41

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