靶向survivin的siRNA及其改性穿膜肽納米給藥系統(tǒng)的研究
發(fā)布時(shí)間：2018-07-29 18:35
【摘要】：腫瘤是嚴(yán)重威脅人類健康的一類疾病,據(jù)世界衛(wèi)生組織報(bào)告,全球新確診和死于腫瘤的人數(shù)都在逐年增加,腫瘤的治療已引起全世界的廣泛關(guān)注。小干擾核糖核酸(Small interfering RNA,si RNA)是通過誘導(dǎo)核糖核酸干擾(RNA interference,RNAi)效應(yīng),在細(xì)胞質(zhì)激發(fā)與之互補(bǔ)的目標(biāo)信使核糖核酸(Messenger RNA,m RNA)沉默,進(jìn)而調(diào)節(jié)蛋白的表達(dá),為腫瘤治療提供了一種有前景的手段。但是,si RNA本身易被核酸酶降解,所以穩(wěn)定性較差;同時(shí)si RNA的親水性強(qiáng)且?guī)ж?fù)電,所以不易透過帶負(fù)電荷的細(xì)胞膜進(jìn)入細(xì)胞質(zhì),最終導(dǎo)致難以發(fā)揮高效的RNAi作用;因此缺乏高效的si RNA以及能夠有效結(jié)合、保護(hù)并遞送si RNA載體的現(xiàn)狀嚴(yán)重限制了其應(yīng)用。本論文合成了一條經(jīng)2′-甲氧基修飾的si RNA序列,對其RNA干擾效果進(jìn)行評價(jià);同時(shí)在穿膜肽八聚精氨酸的基礎(chǔ)上合成了多種脂肪酸修飾的八聚精氨酸,經(jīng)過一系列的試驗(yàn)篩選出能夠高效結(jié)合、保護(hù)并遞送si RNA的改性穿膜肽納米粒;為了克服改性穿膜肽特異性差、靶向配體穿膜效率低的缺點(diǎn),應(yīng)用篩選出的改性穿膜肽和靶向配體制備多功能脂質(zhì)體用于遞送si RNA,并考察其體內(nèi)外遞送si RNA的效果。論文具體內(nèi)容概括如下:1.si RNA在腫瘤細(xì)胞中RNAi效果評價(jià)設(shè)計(jì)了一條靶向survivin基因的2′-甲氧基修飾的si RNA序列,通過實(shí)時(shí)熒光定量PCR、Western blot、流式細(xì)胞術(shù)等方法檢測了si RNA的活性,結(jié)果顯示20 n M的si RNA作用72 h對survivin m RN A的表達(dá)抑制率達(dá)到了90%,對蛋白的表達(dá)抑制率也達(dá)到了80%以上;40 n M的si RNA能夠有效地抑制腫瘤細(xì)胞的增殖,將細(xì)胞周期阻滯在G2/M期,使細(xì)胞凋亡率達(dá)到40%以上,而且能夠?qū)е露喾N細(xì)胞凋亡蛋白表達(dá)量和線粒體膜電位的變化。2.改性穿膜肽的合成、鑒定及細(xì)胞毒性評價(jià)通過固相肽合成的方法,分別將4種脂肪酸(辛酸、硬脂酸、油酸和亞油酸)共價(jià)結(jié)合到八聚精氨酸分子鏈游離氨基末端,制備兩親性的穿膜肽,并利用高效液相色譜法(HPLC)和基質(zhì)輔助激光解吸電離飛行時(shí)間質(zhì)譜(MALDI-TOF-MS)對其進(jìn)行分離純化和鑒定。通過MTT試驗(yàn)對改性穿膜肽進(jìn)行細(xì)胞毒性評價(jià),結(jié)果表明各組改性穿膜肽的細(xì)胞毒性相對較低,可以作為遞送藥物的載體材料。3.改性穿膜肽納米粒的制備及遞送si RNA的研究改性穿膜肽自身具備兩親性,能夠形成自組裝體系,同時(shí)帶有正電荷,可以通過靜電相互作用結(jié)合si RNA形成載si RNA的納米粒,通過對其性質(zhì)表征、細(xì)胞攝取效果和沉默靶基因能力等的研究,篩選出對si RNA結(jié)合效率高、保護(hù)能力強(qiáng)、遞送效果好的改性穿膜肽納米粒。各試驗(yàn)結(jié)果顯示,OA-R8(油酸修飾的八聚精氨酸)納米粒與si RNA的電荷比為1:1時(shí),就可以結(jié)合全部的si RNA,具有最高的結(jié)合效率;在電荷比為4:1,具有良好的血清和聚陰離子穩(wěn)定性;OA-R8載si RNA納米粒的平均粒徑為191.9±17.2 nm,Zeta電位為13.2±5.4 m V,粒徑及電位分布均勻、穩(wěn)定;體外細(xì)胞試驗(yàn)結(jié)果顯示,通過網(wǎng)格蛋白和肌動(dòng)蛋白介導(dǎo)的內(nèi)吞作用,OA-R8載si RNA納米粒能夠成功地將si RNA轉(zhuǎn)染進(jìn)入到腫瘤細(xì)胞內(nèi),將Hep G2和A549細(xì)胞中survivin m RN A的表達(dá)量降低至30.2%和38.9%,survivin蛋白的表達(dá)量降低至42.7%和54.6%,說明改性穿膜肽OA-R8能夠有效地穿過細(xì)胞膜,遞送si RNA進(jìn)入到腫瘤細(xì)胞質(zhì)中,發(fā)揮高效的RNAi作用,進(jìn)而抑制腫瘤細(xì)胞的增殖、阻滯細(xì)胞周期以及促進(jìn)細(xì)胞凋亡等。4.基于改性穿膜肽的多功能脂質(zhì)體的制備及體內(nèi)外抗腫瘤研究雖然改性穿膜肽細(xì)胞毒性小、穿膜效果好,能高效地遞送si RNA,但是改性穿膜肽缺少細(xì)胞特異性,不能很好地靶向到腫瘤組織,且?guī)в写罅空姾?易被血液快速清除。因此本論文將轉(zhuǎn)鐵蛋白(Transferrin,Tf)作為靶向配體,改性穿膜肽OA-R8作為陽離子配體,共同修飾到脂質(zhì)體表面制成Tf和OA-R8雙修飾的多功能脂質(zhì)體(TOLP),裝載survivin si RNA后制成載si RNA的多功能脂質(zhì)體(s TOLP)。s TOLP利用Tf高度的腫瘤細(xì)胞特異性和OA-R8較強(qiáng)的穿膜能力,可以靶向到腫瘤細(xì)胞,穿透細(xì)胞膜,可以將其用于體內(nèi)外輸送si RNA進(jìn)行抗腫瘤的研究。s TOLP帶有正電荷約3.4±2.3 m V,平均粒徑在150.5±14.6 nm,粒度分布均勻,形狀多為規(guī)則的球形。體外細(xì)胞試驗(yàn)結(jié)果表明,多功能脂質(zhì)體對人正常細(xì)胞和腫瘤細(xì)胞的活力無影響,說明脂質(zhì)體中引入改性穿膜肽OA-R8不會(huì)產(chǎn)生任何細(xì)胞毒性;引入OA-R8的載si RNA脂質(zhì)體(s TOLP和OA-R8修飾的脂質(zhì)體s OLP)與未經(jīng)修飾的脂質(zhì)體(s LP)和單獨(dú)用Tf修飾的脂質(zhì)體(s TLP)相比,細(xì)胞攝取效果更好,激光共聚焦觀察細(xì)胞內(nèi)化也得到了相同的結(jié)果;因此OA-R8的引入能夠在體外提高脂質(zhì)體向細(xì)胞內(nèi)遞送si RNA的能力,并且無任何毒副作用。體內(nèi)的試驗(yàn)中,人肝癌異種移植裸小鼠經(jīng)尾靜脈注射脂質(zhì)體,通過監(jiān)測腫瘤體積、小鼠體重等變化,發(fā)現(xiàn)s TOLP組小鼠肝腫瘤的生長受到顯著抑制,與生理鹽水組相比腫瘤抑制率達(dá)到了61.7%;小動(dòng)物活體成像及激光共聚焦觀察si RNA在荷瘤小鼠體內(nèi)的組織分布,發(fā)現(xiàn)經(jīng)s TOLP和s TLP遞送的si RNA主要蓄積在肝臟和腫瘤組織,s TOLP遞送的到達(dá)腫瘤部位的si RNA比s TLP更多;經(jīng)病理切片分析各組織器官的病理變化,發(fā)現(xiàn)s TOLP組小鼠的主要器官與對照組小鼠相比,沒有明顯的組織差異和病變,說明s TOLP在小鼠體內(nèi)沒有全身毒性,可用于體內(nèi)遞送si RNA進(jìn)而抑制腫瘤的生長。綜上所述,本論文合成的靶向survivin的si RNA序列能夠在腫瘤細(xì)胞中發(fā)揮高效的RNAi作用,顯著地抑制survivin基因的表達(dá);合成的改性穿膜肽OA-R8具有低毒性,自組裝形成的納米粒對si RNA的結(jié)合效率高、保護(hù)能力強(qiáng)、遞送效果好,提高了si RNA的穩(wěn)定性和入胞效率;制備的OA-R8和Tf共同修飾的載si RNA多功能脂質(zhì)體,克服了改性穿膜肽特異性差、轉(zhuǎn)鐵蛋白的穿膜效率低的缺點(diǎn),可以跨越體內(nèi)重重屏障將si RNA成功地遞送到腫瘤組織,發(fā)揮高效的RNA干擾作用,最終達(dá)到抑制腫瘤生長的效果。
[Abstract]:Cancer is a kind of disease that seriously threatens human health. According to the WHO, the number of new diagnosis and deaths in the world is increasing year by year. The treatment of cancer has attracted worldwide attention. The Small interfering RNA (Si RNA) is a RNA interference, RNAi effect. Messenger RNA (m RNA) is silent in cytoplasm excitation and complementation, and then regulates the expression of protein, which provides a promising means for cancer treatment. However, Si RNA itself is easily degraded by nuclease, so the stability is poor; meanwhile, Si RNA is highly hydrophilic and negative, so it is not easy to pass the negative charge. The cell membrane enters the cytoplasm and eventually leads to the difficult to play an efficient RNAi effect. Therefore, the lack of efficient Si RNA and the effective binding, and the protection and delivery of Si RNA carriers have severely restricted its application. This paper synthesizes a 2 '- methoxy Modified Si RNA sequence to evaluate the effect of RNA interference; at the same time, the membrane peptide is a membrane peptide. On the basis of eight polyarginine, a variety of fatty acid modified eight polyarginine was synthesized. After a series of experiments, a modified membrane peptide nanoparticle, which could be efficiently combined and protected and delivered Si RNA, was screened. In order to overcome the shortcomings of the modified membrane peptide specificity and the low membrane efficiency of the target ligand, the modified membrane peptide and target matching were used. The multifunctional liposomes were prepared to deliver Si RNA and investigate the effect of delivery of Si RNA in vivo and in vitro. The specific content of the paper is summarized as follows: 1.si RNA designed a Si RNA sequence of 2 '- methoxy modified by the target survivin gene in the tumor cells, and by real time fluorescent quantitative PCR, Western, and flow cytometry. The activity of Si RNA was detected by the method. The results showed that the inhibition rate of the expression of Si RNA of 20 N M to survivin m RN A was 90%, and the inhibition rate of protein expression reached 80%. 40 n can effectively inhibit the proliferation of tumor cells. The synthesis, identification and cytotoxicity evaluation of.2. modified transmembrane peptides that lead to the changes in the expression of a variety of apoptotic protein and mitochondrial membrane potential, combined with the method of solid phase peptide synthesis, combined 4 fatty acids (octanoic acid, stearic acid, oleic acid and linoleic acid) together to the free amino terminal of eight polyarginine chain, and prepare two Pro membrane peptides. It was purified and identified by high performance liquid chromatography (HPLC) and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The cytotoxicity of modified transmembrane peptides was evaluated by MTT test. The results showed that the cytotoxicity of the modified membrane peptides was relatively low and could be used as a carrier material.3. for delivery drugs. Preparation of modified membrane peptide nanoparticles and delivery of Si RNA, the modified membrane peptide has two affinity, which can form a self-assembly system with positive charge, and can form Si RNA loaded Si nanoparticles through electrostatic interaction and Si RNA. By the characterization of its properties, cell uptake and silencing target gene ability and so on, The modified transmembrane peptide nanoparticles with high efficiency, high protection ability and good delivery effect were selected for Si RNA. The results showed that when the charge ratio of OA-R8 (oleic acid modified eight arginine) nanoparticles and Si RNA was 1:1, the whole Si RNA could be combined with the highest binding efficiency, and the charge ratio was 4:1, with good serum and poly Yin. The average particle size of OA-R8 loaded Si RNA nanoparticles was 191.9 + 17.2 nm, the Zeta potential was 13.2 + 5.4 m V, and the particle size and potential distribution were uniform and stable. In vitro cell test results showed that OA-R8 loaded Si RNA nanoparticles could successfully transfect Si RNA into tumor cells through the endocytosis mediated by gridin and actin. The expression of survivin m RN A in Hep G2 and A549 cells was reduced to 30.2% and 38.9%, and the expression of survivin protein was reduced to 42.7% and 54.6%. It indicated that the modified membrane peptide OA-R8 could effectively pass through the cell membrane and send Si RNA into the cytoplasm of the tumor, and play an efficient RNAi action, and then inhibit the proliferation of tumor cells and block the cell cycle. As well as the preparation of.4. based multifunctional liposomes based on modified membrane peptide and the antitumor research in vivo and in vivo and in vivo, although the modified transmembrane peptide has small toxicity, good membrane effect and efficient delivery of Si RNA, the modified membrane peptide lacks cell specificity and can not target tumor tissue well, with a large number of positive charges and easy to be used. So in this paper, Transferrin (Tf) is used as a target ligand and modified membrane peptide OA-R8 is used as a cationic ligand, which is modified to the surface of liposome to make Tf and OA-R8 double modified multifunctional liposomes (TOLP). After loading survivin Si RNA, the Si RNA multi-functional liposome is made. The specificity of tumor cells and the strong membrane ability of OA-R8 can target the tumor cells and penetrate the cell membrane. It can be used to transport Si RNA in vivo and in vivo for anti-tumor research..s TOLP has a positive charge of about 3.4 + 2.3 m V, the average particle size is 150.5 + 14.6 nm, the particle size distribution is uniform, and the shape is mostly regular. In vitro cell test junction The results showed that the multifunctional liposomes had no effect on the activity of human normal cells and tumor cells, indicating that the introduction of modified membrane peptide OA-R8 in liposomes would not produce any cytotoxicity; the introduction of the OA-R8 loaded Si RNA liposomes (s TOLP and OA-R8 modified liposomes s OLP) and the unmodified liposomes (s LP) and the liposomes modified by Tf alone In contrast, cell uptake is better, and laser confocal observation of cell internalization is also the same. Therefore, the introduction of OA-R8 can improve the ability of liposomes to deliver Si RNA into cells in vitro, without any toxic and side effects. In the test of human liver cancer xenotransplantation, nude mice were injected with liposomes through the tail vein, by monitoring the swelling. The tumor volume, mouse weight and other changes showed that the growth of liver tumor in the s TOLP group was significantly inhibited, and the tumor inhibition rate was 61.7% compared with the normal saline group. The tissue distribution of Si RNA in the tumor bearing mice was observed by the living body imaging and laser confocal microscopy, and the Si RNA delivered by s TOLP and s TLP was mainly accumulated in the liver and swelling. The Si RNA delivered by s TOLP was more than that of s TLP. The pathological changes of tissues and organs were analyzed by pathological section, and the main organs of the s TOLP mice were found to have no obvious histological differences and pathological changes compared with the control group, indicating that s TOLP had no systemic toxicity in mice, and could be used in the delivery of Si RNA in the body. To sum up the growth of the tumor, the Si RNA sequence of the target survivin, synthesized in this paper, can play an efficient RNAi role in the tumor cells and significantly inhibit the expression of the survivin gene; the synthesized modified transmembrane peptide OA-R8 has low toxicity, and the self assembled nanoparticles have high binding efficiency to Si RNA, strong protection and good delivery effect. The stability and cell efficiency of Si RNA were improved; the Si RNA multifunctional liposomes, which were modified by OA-R8 and Tf, overcome the shortcomings of the modified transmembrane peptide specificity and the low membrane efficiency of the transferrin, which could be successfully delivered to the swelling tumor tissue across the barrier of the body, and the effective RNA interference was achieved. Finally, the inhibition of Si RNA was achieved. The effect of the growth of the tumor.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R943
本文編號：2153626
[Abstract]:Cancer is a kind of disease that seriously threatens human health. According to the WHO, the number of new diagnosis and deaths in the world is increasing year by year. The treatment of cancer has attracted worldwide attention. The Small interfering RNA (Si RNA) is a RNA interference, RNAi effect. Messenger RNA (m RNA) is silent in cytoplasm excitation and complementation, and then regulates the expression of protein, which provides a promising means for cancer treatment. However, Si RNA itself is easily degraded by nuclease, so the stability is poor; meanwhile, Si RNA is highly hydrophilic and negative, so it is not easy to pass the negative charge. The cell membrane enters the cytoplasm and eventually leads to the difficult to play an efficient RNAi effect. Therefore, the lack of efficient Si RNA and the effective binding, and the protection and delivery of Si RNA carriers have severely restricted its application. This paper synthesizes a 2 '- methoxy Modified Si RNA sequence to evaluate the effect of RNA interference; at the same time, the membrane peptide is a membrane peptide. On the basis of eight polyarginine, a variety of fatty acid modified eight polyarginine was synthesized. After a series of experiments, a modified membrane peptide nanoparticle, which could be efficiently combined and protected and delivered Si RNA, was screened. In order to overcome the shortcomings of the modified membrane peptide specificity and the low membrane efficiency of the target ligand, the modified membrane peptide and target matching were used. The multifunctional liposomes were prepared to deliver Si RNA and investigate the effect of delivery of Si RNA in vivo and in vitro. The specific content of the paper is summarized as follows: 1.si RNA designed a Si RNA sequence of 2 '- methoxy modified by the target survivin gene in the tumor cells, and by real time fluorescent quantitative PCR, Western, and flow cytometry. The activity of Si RNA was detected by the method. The results showed that the inhibition rate of the expression of Si RNA of 20 N M to survivin m RN A was 90%, and the inhibition rate of protein expression reached 80%. 40 n can effectively inhibit the proliferation of tumor cells. The synthesis, identification and cytotoxicity evaluation of.2. modified transmembrane peptides that lead to the changes in the expression of a variety of apoptotic protein and mitochondrial membrane potential, combined with the method of solid phase peptide synthesis, combined 4 fatty acids (octanoic acid, stearic acid, oleic acid and linoleic acid) together to the free amino terminal of eight polyarginine chain, and prepare two Pro membrane peptides. It was purified and identified by high performance liquid chromatography (HPLC) and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The cytotoxicity of modified transmembrane peptides was evaluated by MTT test. The results showed that the cytotoxicity of the modified membrane peptides was relatively low and could be used as a carrier material.3. for delivery drugs. Preparation of modified membrane peptide nanoparticles and delivery of Si RNA, the modified membrane peptide has two affinity, which can form a self-assembly system with positive charge, and can form Si RNA loaded Si nanoparticles through electrostatic interaction and Si RNA. By the characterization of its properties, cell uptake and silencing target gene ability and so on, The modified transmembrane peptide nanoparticles with high efficiency, high protection ability and good delivery effect were selected for Si RNA. The results showed that when the charge ratio of OA-R8 (oleic acid modified eight arginine) nanoparticles and Si RNA was 1:1, the whole Si RNA could be combined with the highest binding efficiency, and the charge ratio was 4:1, with good serum and poly Yin. The average particle size of OA-R8 loaded Si RNA nanoparticles was 191.9 + 17.2 nm, the Zeta potential was 13.2 + 5.4 m V, and the particle size and potential distribution were uniform and stable. In vitro cell test results showed that OA-R8 loaded Si RNA nanoparticles could successfully transfect Si RNA into tumor cells through the endocytosis mediated by gridin and actin. The expression of survivin m RN A in Hep G2 and A549 cells was reduced to 30.2% and 38.9%, and the expression of survivin protein was reduced to 42.7% and 54.6%. It indicated that the modified membrane peptide OA-R8 could effectively pass through the cell membrane and send Si RNA into the cytoplasm of the tumor, and play an efficient RNAi action, and then inhibit the proliferation of tumor cells and block the cell cycle. As well as the preparation of.4. based multifunctional liposomes based on modified membrane peptide and the antitumor research in vivo and in vivo and in vivo, although the modified transmembrane peptide has small toxicity, good membrane effect and efficient delivery of Si RNA, the modified membrane peptide lacks cell specificity and can not target tumor tissue well, with a large number of positive charges and easy to be used. So in this paper, Transferrin (Tf) is used as a target ligand and modified membrane peptide OA-R8 is used as a cationic ligand, which is modified to the surface of liposome to make Tf and OA-R8 double modified multifunctional liposomes (TOLP). After loading survivin Si RNA, the Si RNA multi-functional liposome is made. The specificity of tumor cells and the strong membrane ability of OA-R8 can target the tumor cells and penetrate the cell membrane. It can be used to transport Si RNA in vivo and in vivo for anti-tumor research..s TOLP has a positive charge of about 3.4 + 2.3 m V, the average particle size is 150.5 + 14.6 nm, the particle size distribution is uniform, and the shape is mostly regular. In vitro cell test junction The results showed that the multifunctional liposomes had no effect on the activity of human normal cells and tumor cells, indicating that the introduction of modified membrane peptide OA-R8 in liposomes would not produce any cytotoxicity; the introduction of the OA-R8 loaded Si RNA liposomes (s TOLP and OA-R8 modified liposomes s OLP) and the unmodified liposomes (s LP) and the liposomes modified by Tf alone In contrast, cell uptake is better, and laser confocal observation of cell internalization is also the same. Therefore, the introduction of OA-R8 can improve the ability of liposomes to deliver Si RNA into cells in vitro, without any toxic and side effects. In the test of human liver cancer xenotransplantation, nude mice were injected with liposomes through the tail vein, by monitoring the swelling. The tumor volume, mouse weight and other changes showed that the growth of liver tumor in the s TOLP group was significantly inhibited, and the tumor inhibition rate was 61.7% compared with the normal saline group. The tissue distribution of Si RNA in the tumor bearing mice was observed by the living body imaging and laser confocal microscopy, and the Si RNA delivered by s TOLP and s TLP was mainly accumulated in the liver and swelling. The Si RNA delivered by s TOLP was more than that of s TLP. The pathological changes of tissues and organs were analyzed by pathological section, and the main organs of the s TOLP mice were found to have no obvious histological differences and pathological changes compared with the control group, indicating that s TOLP had no systemic toxicity in mice, and could be used in the delivery of Si RNA in the body. To sum up the growth of the tumor, the Si RNA sequence of the target survivin, synthesized in this paper, can play an efficient RNAi role in the tumor cells and significantly inhibit the expression of the survivin gene; the synthesized modified transmembrane peptide OA-R8 has low toxicity, and the self assembled nanoparticles have high binding efficiency to Si RNA, strong protection and good delivery effect. The stability and cell efficiency of Si RNA were improved; the Si RNA multifunctional liposomes, which were modified by OA-R8 and Tf, overcome the shortcomings of the modified transmembrane peptide specificity and the low membrane efficiency of the transferrin, which could be successfully delivered to the swelling tumor tissue across the barrier of the body, and the effective RNA interference was achieved. Finally, the inhibition of Si RNA was achieved. The effect of the growth of the tumor.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R943
【相似文獻(xiàn)】
相關(guān)期刊論文 前5條
1 莊煒東;阮世龍;楊旭中;王佳瑋;王富軍;趙健;;一種高效人源性穿膜肽的生物活性研究[J];食品與藥品;2013年02期
2 李莉蓉;施用暉;樂國偉;;來自穿膜肽的新肽的抗菌活性及抑菌機(jī)制[J];微生物學(xué)報(bào);2013年09期
3 張楠;白銀;趙景壯;葉賢龍;王文飛;任桂萍;李德山;荊燕;;多聚精氨酸融合增強(qiáng)型綠色熒光蛋白制備方法及穿膜效果[J];生物工程學(xué)報(bào);2013年11期
4 王軍;雷楗勇;陳蘊(yùn);金堅(jiān);;細(xì)胞穿膜肽TAT與小鼠干細(xì)胞轉(zhuǎn)錄因子Oct4融合表達(dá)及純化[J];食品與生物技術(shù)學(xué)報(bào);2013年09期
5 ;[J];;年期
相關(guān)會(huì)議論文 前7條
1 王小波;周明;費(fèi)浩;;一種基于環(huán)金屬銥化合物與組氨酸結(jié)合的新型多肽標(biāo)記方式及用于細(xì)胞穿膜肽的成像研究[A];中國化學(xué)會(huì)第27屆學(xué)術(shù)年會(huì)第03分會(huì)場摘要集[C];2010年
2 王宏博;楊建海;王瑋;劉文廣;;富含精氨酸的細(xì)胞穿膜肽的研究進(jìn)展(綜述)[A];天津市生物醫(yī)學(xué)工程學(xué)會(huì)第三十一屆學(xué)術(shù)年會(huì)論文集[C];2011年
3 王怡;肖健;林海環(huán);楊靜;李校X;;新穿膜肽蛋白His-T1-GFP的跨膜效率和細(xì)胞毒性研究[A];2007年全國生化與生物技術(shù)藥物學(xué)術(shù)年會(huì)論文集[C];2007年
4 王琥;柳長柏;黃宇彬;;不同條件下細(xì)胞穿膜肽TAT的穿膜效率的比較[A];第二屆中國醫(yī)學(xué)細(xì)胞生物學(xué)學(xué)術(shù)大會(huì)暨細(xì)胞生物學(xué)教學(xué)改革會(huì)議論文集[C];2008年
5 薛改青;祖莉莉;;穿膜肽Transportan10氣相解離動(dòng)力學(xué)的研究[A];第十三屆全國化學(xué)動(dòng)力學(xué)會(huì)議報(bào)告摘要集[C];2013年
6 馬巖巖;榮靖;彭鑫磊;劉秋英;舒輝萍;陳海佳;王一飛;任哲;;穿膜肽KGF-2的構(gòu)建及其對HaCaT增殖作用的影響[A];中國遺傳學(xué)會(huì)“第十一屆全國激光生物學(xué)學(xué)術(shù)會(huì)議”暨《激光生物學(xué)報(bào)》創(chuàng)刊廿周年慶祝會(huì)、第十屆粵港生物物理研討會(huì)暨2012年廣東生物物理學(xué)術(shù)年會(huì)會(huì)議資料[C];2012年
7 張俊萍;孫青山;賈原;;穿膜肽介導(dǎo)p53多肽抗原致敏的DC的體外抗瘤效應(yīng)研究[A];中國腫瘤內(nèi)科進(jìn)展 中國腫瘤醫(yī)師教育（2014）[C];2014年
相關(guān)博士學(xué)位論文 前4條
1 馬嚴(yán);可在體實(shí)現(xiàn)胞內(nèi)直接投遞的新型細(xì)胞穿膜肽的構(gòu)建、篩選和穿膜機(jī)制研究[D];華中科技大學(xué);2012年
2 李玉環(huán);靶向survivin的siRNA及其改性穿膜肽納米給藥系統(tǒng)的研究[D];吉林大學(xué);2016年
3 嚴(yán)世榮;穿膜肽攜帶的目的蛋白穿膜效應(yīng)的研究[D];華中科技大學(xué);2006年
4 李珂;靶向toll樣受體2多肽的篩選、鑒定及應(yīng)用[D];北京協(xié)和醫(yī)學(xué)院;2013年
相關(guān)碩士學(xué)位論文 前10條
1 荊鑫鑫;高效重組穿膜肽-vp3的篩選及其對Hela細(xì)胞的抑制作用[D];佳木斯大學(xué);2015年
2 劉暢;穿膜肽介導(dǎo)的眼內(nèi)基因遞送系統(tǒng)[D];復(fù)旦大學(xué);2014年
3 馬德云;細(xì)胞穿膜肽Pep-1和人表皮生長因子在大腸桿菌中的重組表達(dá)[D];天津科技大學(xué);2014年
4 莊煒東;新型人源性穿膜肽機(jī)理研究及性質(zhì)比較[D];華東理工大學(xué);2013年
5 楊冬梅;富含胍基的細(xì)胞穿膜肽合成[D];華中科技大學(xué);2012年
6 張宏達(dá);穿膜肽介導(dǎo)的新型基因轉(zhuǎn)運(yùn)載體的研究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2008年
7 韋一凡;細(xì)胞穿膜肽在藻類細(xì)胞外源物質(zhì)轉(zhuǎn)導(dǎo)中的應(yīng)用[D];中國科學(xué)院研究生院（海洋研究所）;2014年
8 劉云龍;穿膜肽-β-葡萄糖苷酶—西妥昔單抗偶聯(lián)物的制備與鑒定[D];廣西醫(yī)科大學(xué);2015年
9 吳強(qiáng);穿膜肽/NF-κB拮抗多肽原核表達(dá)載體的構(gòu)建、表達(dá)、純化與功能檢測[D];安徽農(nóng)業(yè)大學(xué);2006年
10 王光文;穿膜肽-β-葡萄糖苷酶偶聯(lián)物制備鑒定及激活苦杏仁苷殺傷膀胱癌細(xì)胞的初步研究[D];廣西醫(yī)科大學(xué);2014年
，本文編號：2153626
本文鏈接：http://sikaile.net/yixuelunwen/yiyaoxuelunwen/2153626.html
最近更新
教材專著
