本文選題：Rab27a　切入點(diǎn)：腦膠質(zhì)瘤　出處：《安徽醫(yī)科大學(xué)》2014年博士論文

【摘要】：目的:探討Rab27a的表達(dá)對(duì)人腦膠質(zhì)瘤U251細(xì)胞生長(zhǎng)、增殖、凋亡及侵襲遷移等生物學(xué)行為的影響,為將來人腦膠質(zhì)瘤的生物治療提供實(shí)驗(yàn)基礎(chǔ)。方法:構(gòu)建pc DNA3.1-Rab27a真核重組表達(dá)載體,利用脂質(zhì)體將pc DNA3.1-Rab27a和Rab27a-sh RNA轉(zhuǎn)染U251,Western blot檢測(cè)Rab27a蛋白的表達(dá);MTT法檢測(cè)細(xì)胞活力;流式細(xì)胞術(shù)檢測(cè)Rab27a異位表達(dá)并分析其對(duì)U251細(xì)胞增殖、凋亡的影響;Transwell侵襲小室等方法檢測(cè)U251細(xì)胞侵襲性;通過檢測(cè)Rab27a的異位表達(dá),分析對(duì)組織蛋白酶D分泌的影響;應(yīng)用熒光素酶報(bào)告基因系統(tǒng)驗(yàn)證Rab27a是mi R-124的靶基因。結(jié)果:Western blot結(jié)果顯示:與對(duì)照組相比,pc DNA3.1-Rab27a轉(zhuǎn)染組Rab27a的表達(dá)顯著增加(p0.01),而Rab27a-sh RNA轉(zhuǎn)染組Rab27a的表達(dá)顯著降低(p0.01)。MTT、流式細(xì)胞術(shù)和Transwell侵襲小室檢測(cè)結(jié)果顯示:與對(duì)照組相比,pc DNA3.1-Rab27a轉(zhuǎn)染組細(xì)胞活力(p0.01)、增殖指數(shù)(p0.05)、遷移能力顯著增加(p0.01),Rab27a-sh RNA轉(zhuǎn)染組細(xì)胞活力(p0.01)、增殖指數(shù)(p0.05)、遷移能力顯著降低(p0.01)。流式細(xì)胞術(shù)分析凋亡過程,結(jié)果顯示:與對(duì)照組相比,pc DNA3.1-Rab27a轉(zhuǎn)染組細(xì)胞凋亡降低(p0.05),而Rab27a-sh RNA轉(zhuǎn)染組細(xì)胞凋亡增加(p0.05)。組織蛋白酶D檢測(cè)結(jié)果顯示:與正常對(duì)照組相比,pc DNA3.1-Rab27a轉(zhuǎn)染組中組織蛋白酶D的分泌量高于正常對(duì)照組(p0.05)。結(jié)論:Rab27a基因具有促進(jìn)腦膠質(zhì)瘤細(xì)胞U251細(xì)胞的活力、增殖、遷移,并抑制其凋亡作用,其作用機(jī)制可能通過促進(jìn)組織蛋白酶D的分泌,和受mi R-124的調(diào)控來發(fā)揮作用。抑制Rab27a表達(dá)的治療策略有望使膠質(zhì)瘤患者受益。目的:根據(jù)Rab27a在腦膠質(zhì)瘤組織中的表達(dá)情況在老年腦膠質(zhì)瘤患者中進(jìn)行個(gè)體化化療,觀察其治療效果、不良反應(yīng)和生存期。方法:入組時(shí)間:2011年7月至2013年5月。年齡≥65歲;經(jīng)手術(shù)切除或活檢后的組織病理學(xué)確診的腦膠質(zhì)瘤患者,測(cè)定組織Rab27a表達(dá)水平。Rab27a表達(dá)者設(shè)為治療組,應(yīng)用單藥替莫唑胺化療,治療劑量:150~200mg/m2,口服,每日一次,d1~5;每28天為一周期,至少治療2周期。Rab27a不表達(dá)者設(shè)為對(duì)照組,僅對(duì)癥處理,未進(jìn)行化療。按照WHO標(biāo)準(zhǔn)評(píng)價(jià)療效、CTCAE 4.0標(biāo)準(zhǔn)評(píng)價(jià)藥物不良反應(yīng),其它觀察指標(biāo)為OS和PFS。結(jié)果:按照入組標(biāo)準(zhǔn),治療組共有32例患者進(jìn)入臨床研究;中位年齡70歲,中位KPS:70分。可評(píng)價(jià)病例29例,其中9例PR(31%)、12例SD(41%)、8例PD(28%)。自診斷開始的中位OS為7.4個(gè)月,中位PFS為6.1個(gè)月。所有入組患者共進(jìn)行了160周期單藥替莫唑胺化療,中位為4周期。不良反應(yīng)以Ⅰ~Ⅱ度為主,包括:惡心、嘔吐、疲勞、便秘、骨髓毒性等。Ⅲ~Ⅳ度中性粒細(xì)胞減少的出現(xiàn)率為9.38%(3/32)患者,Ⅳ度血小板下降的出現(xiàn)率為6.25%(2/32)。對(duì)照組共入組21例,中位年齡68歲,中位KPS:70分,自診斷開始的中位OS為5.7個(gè)月,中位PFS為4.0個(gè)月。結(jié)論:Rab27a指導(dǎo)下的替莫唑胺化療在腦膠質(zhì)瘤有廣闊的應(yīng)用前景,替莫唑胺優(yōu)點(diǎn)在于:療效顯著;大多數(shù)副作用可耐受;口服給藥方便;改善其生活質(zhì)量。Rab27a對(duì)患者生存期的預(yù)測(cè)價(jià)值有待進(jìn)一步研究。
[Abstract]:Objective: To investigate the growth and proliferation of U251 human glioma cells Rab27a expression, apoptosis and invasion influence the biological behavior of migration, to provide the experimental basis for the biological treatment of glioma in the future. Methods: to construct the PC DNA3.1-Rab27a eukaryotic expression vector by liposome PC DNA3.1-Rab27a and Rab27a-sh RNA U251 transfection, the expression of Rab27a protein detection of Western blot; the cell viability was measured by MTT; flow cytometry analysis of ectopic expression of Rab27a and the proliferation of U251 cells, apoptosis; Transwell assay detected U251 cell invasion; by ectopic expression of Rab27a analysis of effect on secretion of cathepsin D using luciferase reporter system verification; Rab27a is the target gene mi R-124. Results: Western blot showed that: compared with the control group, the expression of PC DNA3.1-Rab27a transfection group Rab27a increased significantly Plus (P0.01), and the expression of Rab27a-sh RNA transfection group significantly decreased Rab27a (P0.01).MTT, flow cytometry and Transwell assay. The results showed that: compared with the control group, PC transfection group DNA3.1-Rab27a cell viability (P0.01), proliferation index (P0.05), the migration ability was significantly increased (P0.01), Rab27a-sh RNA transfection activity (P0.01), proliferation index (P0.05), the migration ability decreased significantly (P0.01). Analysis of the process of apoptosis, flow cytometry results showed that: compared with the control group, the apoptosis of PC cells in DNA3.1-Rab27a transfection group decreased (P0.05), and the apoptosis of Rab27a-sh cells in RNA transfection group increased (P0.05). The detection of cathepsin D the results showed that: compared with normal control group, PC secretion in DNA3.1-Rab27a transfection group of cathepsin D is higher than the normal control group (P0.05). Conclusion: Rab27a gene can promote glioma cell viability of U251 cells, proliferation, migration, and suppression The role of apoptosis, its mechanism may promote the secretion of cathepsin D, MI and R-124 of the control to play the role of therapeutic strategies to inhibit the expression of Rab27a is expected to make the glioma patients. Objective: according to the expression of Rab27a in glioma tissues of individual chemotherapy in elderly patients with cerebral glioma, observation the therapeutic effect, adverse reaction and survival. Methods: in the group of time: from July 2011 to May 2013. 65 years of age or older; the patients with glioma confirmed histological pathology after surgical excision or biopsy, determination of tissue Rab27a expression levels of.Rab27a expression were divided into treatment group, application of single drug temozolomide that dose: 150~200mg/m2, oral, once daily, d1~5; every 28 days for a period of at least 2 cycles of treatment, the expression of.Rab27a were set as control group, only symptomatic treatment, no chemotherapy. According to WHO evaluation criteria, C TCAE 4.0鏍囧噯璇勪環(huán)鑽墿涓嶈壇鍙嶅簲,鍏跺畠瑙傚療鎸囨爣涓篛S鍜孭FS.緇撴灉:鎸夌収鍏ョ粍鏍囧噯,娌葷枟緇勫叡鏈，

本文編號(hào)：1721047