本文選題：聚酰胺胺型樹枝狀分子 + CXCR��； 參考：《中國腫瘤生物治療雜志》2015年01期

【摘要】：目的:制備聚酰胺胺型樹枝狀分子(polyamidoamine dendrimer,PAMAM-D)載送CXCR4-shRNA的納米復合微粒(PAMAM-shRNA),研究其在體外抑制人腎癌A498細胞增殖的效應。方法:將第7代的PAMAM-D室溫下與CXCR4-shRNA混合(質(zhì)量比為1∶0.73),制備成納米樹形分子與CXCR4-shRNA的納米復合物PAMAM-shRNA,采用透射電鏡觀察PAMAMshRNA的形態(tài)結(jié)構(gòu),激光粒徑儀測定其粒徑。分別以PAMAM-shRNA、CXCR4-shRNA和PAMAM-D轉(zhuǎn)染A498細胞,MTT法檢測PAMAM-shRNA對腎癌細胞A498增殖的抑制作用,流式細胞術(shù)檢測PAMAM-shRNA誘導A498細胞的凋亡情況,Real-time PCR分析轉(zhuǎn)染后腎癌A498細胞CXCR4 mRNA的表達水平。結(jié)果:成功制備的新型納米復合微粒PAMAM-shRNA分散性好,不粘連,其平均粒徑為(176.5±25.48)nm。PAMAM-shRNA可以有效地抑制人腎癌細胞A498的增殖,且隨著PAMAM-shRNA濃度和藥物作用時間的增加,細胞增殖抑制的效果越明顯,其最高增殖抑制率達(66.5±2.7)%;其還可加強誘導腎癌A498細胞凋亡。Real-time PCR檢測結(jié)果表明,與CXCR4-shRNA組相比,PAMAM-shRNA組的CXCR4 mRNA的表達水平明顯下降[(0.29±0.035)vs(0.70±0.084),P0.05]。結(jié)論:PAMAM-D能高效遞送CXCR4-shRNA進入A498細胞,PAMAM-shRNA以劑量和時間依賴方式顯著抑制腎癌細胞增殖和誘導腎癌細胞凋亡,其在腫瘤基因治療中具有潛在的應用價值。
[Abstract]:Aim: to prepare polyamidoamine dendrimer-PAMAM-D nanoparticles (PAMAM-shRNA) carrying CXCR4-shRNA, and to study the inhibitory effect of PAMAM-shRNA on the proliferation of human renal cell carcinoma A498 cells in vitro. Methods: the 7th generation PAMAM-D was mixed with CXCR4-shRNA at room temperature (mass ratio 1: 0.73), and the nanocomposite PAMAM-shRNA was prepared. The morphology of PAMAM-shRNA was observed by transmission electron microscope. The particle size of PAMAMshRNA was measured by laser particle size analyzer. The inhibitory effect of PAMAM-shRNA on the proliferation of renal cancer cell A498 was detected by MTT assay with PAMAM-shRNA-CXCR4-shRNA and PAMAM-D transfected A498 cell line. Flow cytometry was used to detect the apoptosis of A498 cells induced by PAMAM-shRNA. Real-time PCR was used to analyze the expression level of CXCR4 mRNA in A498 cells transfected with PAMAM-shRNA. Results: PAMAM-shRNA, a new type of nano-composite particle, had good dispersity and no adhesion. The average diameter of PAMAM-shRNA was (176.5 鹵25.48) nm.PAMAM-shRNA could effectively inhibit the proliferation of human renal cancer cell A498, and with the increase of PAMAM-shRNA concentration and drug action time, PAMAM-shRNA could effectively inhibit the proliferation of human renal cancer cell line A498. The more obvious the effect of cell proliferation inhibition was, the highest inhibitory rate of cell proliferation was (66.5 鹵2.7), and the results of real-time PCR showed that the expression level of CXCR4 mRNA in PAMAM-shRNA group was significantly lower than that in CXCR4-shRNA group [(0.29 鹵0.035) vs (0.70 鹵0.084) P 0.05], and the expression of CXCR4 mRNA in CXCR4-shRNA group was significantly lower than that in CXCR4-shRNA group [(0.29 鹵0.035) vs (, 0.70 鹵0.084) (P0.05)]. Conclusion: the fraction PAMAM-D can efficiently deliver CXCR4-shRNA into A498 cells and inhibit the proliferation and apoptosis of RCC cells in a dose-and time-dependent manner. It has potential application value in tumor gene therapy.
【作者單位】： 武警上�？傟犪t(yī)院腎內(nèi)科;第二軍醫(yī)大學長海醫(yī)院特需診療科;
【基金】：國家自然科學基金資助項目(No.81202019) 上海市衛(wèi)生局青年科研資助項目(No.2011Y197)~~
【分類號】：R737.11

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