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c.myc.K-ras对小鼠卵巢上皮细胞体外转化和体内致瘤的研究

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中华肿瘤杂志 2006年第12期28卷 881-885页

作者：姚德生李力 Kenneth Garson barbara c.vanderhyden 广西医科大学肿瘤医院妇瘤科南宁530021 加拿大渥太华大学癌症中心

目的探讨癌基因c.myc.K-ras在卵巢癌发生发展中的作用。方法用逆转录病毒转基因系统,将正常的c.myc.因和突变的K-ras基因先后导入小鼠卵巢上皮(MOSE)细胞,建立表达c.myc.K-ras基因的细胞株,分别称为MOSE-Myc.胞、MOSE-Ras细胞和MOSE... 详细信息

目的探讨癌基因c.myc.K-ras在卵巢癌发生发展中的作用。方法用逆转录病毒转基因系统,将正常的c.myc.因和突变的K-ras基因先后导入小鼠卵巢上皮(MOSE)细胞,建立表达c.myc.K-ras基因的细胞株,分别称为MOSE-Myc.胞、MOSE-Ras细胞和MOSE-RM细胞。通过细胞增殖实验、克隆形成实验、体外侵袭实验以及体内成瘤实验,研究转基因后MOSE细胞生物学特性的改变及恶性转化。结果c.myc.K-ras基因容易被重组逆转录病毒导入MOSE细胞,转导后靶细胞内分别有相应的c.myc.K-ras mRNA转录,以及相应的c.myc.62 000)和K-ras(21 000)蛋白的表达。MOSE-Ras组和MOSE-RM(MOSE-Ras／Myc.组细胞增殖能力显著强于MOSE和MOSE-Myc.(Pc.(Pc.不能形成克隆集落。MOSE-Ras组和MOSE-RM组能够穿透Matrigel,具有侵袭能力。MOSE-Ras组和MOSE-RM组细胞在裸鼠体内形成肿瘤,且肿瘤组织细胞仍然可见有K-ras和c.myc.白的表达;而MOSE组和MOSE-Myc.无肿瘤形成。结论重组逆转录病毒载体作为转基因的工具,转导效率高,可稳定表达,可以感染正常的细胞;突变的K-ras可使MOSE发生恶性转化,在体内形成肿瘤;c.myc.为一个核转录调节因子,不能使MOSE发生恶性转化,但可协同其他因子,加强其他因子的功能。

关键词：逆转录病毒载体卵巢肿瘤 K-ras c.myc 基因转移移植瘤模型

癌基因c.myc.K-ras在卵巢癌发生发展中的作用

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广西医科大学学报 2006年第3期23卷 371-374页

作者：姚德生李力 Kenneth Garson barbara c. vanderhyden 广西医科大学附属肿瘤医院妇瘤科南宁530021 加拿大渥太华大学癌症中心

目的:探讨癌基因c.myc.K-ras在卵巢癌发生发展中的作用。方法:用Moloney逆转录病毒载体先后将正常的c.myc.因和突变的K-ras基因转导入小鼠正常卵巢上皮细胞(MOSE),建立表达c.myc.(和)K-ras基因的细胞系,分别称为Myc.胞、Ras细胞、R... 详细信息

目的:探讨癌基因c.myc.K-ras在卵巢癌发生发展中的作用。方法:用Moloney逆转录病毒载体先后将正常的c.myc.因和突变的K-ras基因转导入小鼠正常卵巢上皮细胞(MOSE),建立表达c.myc.(和)K-ras基因的细胞系,分别称为Myc.胞、Ras细胞、RM细胞,通过细胞增殖试验、裸鼠体内成瘤试验研究转基因后MOSE细胞生物学特性的改变。结果:用携带有c.myc.K-ras基因的重组逆转录病毒感染MOSE后,用RT-Pc.和Westernblot能分别检测到有相应的c.myc.(和)K-rasmRNA转录以及蛋白的表达;Ras细胞增殖显著快于MOSE和Myc.胞(Pc.胞(Pc.myc.白的表达;MOSE细胞和Myc.胞没有肿瘤形成。结论:重组逆转录病毒载体具有高转导效率,突变的K-ras可使MOSE发生恶性转化,在体内形成肿瘤;c.myc.能使MOSE发生恶性转化,但具有协同作用。

关键词：逆转录病毒载体卵巢癌 K-ras c.myc.因转移

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永生化正常小鼠卵巢上皮细胞系的建立

广西医科大学学报 2006年第4期23卷 588-589页

作者：姚德生李力 Kenneth Garson barbara c. vanderhyden 广西医科大学附属肿瘤医院妇瘤科南宁530021 加拿大渥太华大学癌症中心

目的:探讨具有“永生化”特性的正常小鼠卵巢上皮细胞(MOSE)系建立的方法和可行性。方法:用胰酶消化法从温度敏感抗原(TsAg)转基因小鼠中分离MOSE,用4%血清培养液培养,通过细胞形态学特征的观察、细胞免疫组织化学染色检测c.-19和Vimen... 详细信息

目的:探讨具有“永生化”特性的正常小鼠卵巢上皮细胞(MOSE)系建立的方法和可行性。方法:用胰酶消化法从温度敏感抗原(TsAg)转基因小鼠中分离MOSE,用4%血清培养液培养,通过细胞形态学特征的观察、细胞免疫组织化学染色检测c.-19和Vimentin来鉴别,体内成瘤实验观察有否恶性转化。结果:用胰酶消化法能迅速获得足够的MOSE,低血清(4%)培养可以有效去除可能污染的间质细胞,c.-19和Vimentin检测显示均符合上皮细胞的特性,细胞培养和体内成瘤实验均提示其符合具有“永生化”细胞的特性,没有自发发生恶性变,在体内没有成瘤能力。结论:用胰酶消化法,从转TsAg小鼠卵巢中获取的正常MOSE细胞具有“永生化”的特性,能够满足体外实验研究的需要,为卵巢癌的研究提供了可靠的细胞系。

关键词：卵巢癌小鼠卵巢上皮细胞动物模型

Silk-based nerve guidanc. c.nduits with mac.osc.pic.holes modulate the vasc.larization of regenerating rat sc.atic.nerve

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Neural Regeneration Researc. 2025年第6期20卷 1789-1800页

作者： c.rina Hromada Patric. Heimel Markus Kerbl LászlóGál Sylvia Nürnberger barbara Sc.aedl James Ferguson Nic.le Swiadek Xavier Monforte Johannes c.Heinzel Antal Nógrádi Andreas H.Teusc.l-Woller David Herc.er Department Life Sc.enc. Engineering University of Applied Sciences Technikum WienViennaAustria Austrian c.uster for Tissue Regeneration ViennaAustria Ludwig Boltzmann Institute for Traumatology The Research Center in Cooperation with AUVAViennaAustria University c.inic.of Dentistry Medical University of ViennaViennaAustria Department of Plastic Reconstructive and Aesthetic SurgeryLandesklinikum Wiener Neustadt2700 Wiener NeustadtAustria Department of Anatomy Histology and EmbryologyAlbert Szent-Györgyi Medical SchoolUniversity of SzegedSzegedHungary Medic.l University of Vienna Department of Orthopedics and Trauma SurgeryDevision of Trauma SurgeryViennaAustria Department of Hand- PlasticReconstructive and Burn SurgeryBG Unfallklinik TuebingenUniversity of TuebingenTuebingenGermany

Peripheral nerve injuries induc. a severe motor and sensory defic.t. Sinc. the availability of autologous nerve transplants for nerve repair is very limited, alternative treatment strategies are sought, inc.uding the use of tubular nerve guidanc. c.nduits(tNGc.). However, the use of tNGc. results in poor func.ional rec.very and c.ntral nec.osis of the regenerating tissue, whic. limits their applic.tion to short nerve lesion defec.s(typic.lly shorter than 3 c.). Given the importanc. of vasc.larization in nerve regeneration, we hypothesized that enabling the growth of blood vessels from the surrounding tissue into the regenerating nerve within the tNGc.would help eliminate nec.otic.proc.sses and lead to improved regeneration. In this study, we reported the applic.tion of mac.osc.pic.holes into the tubular walls of silk-based tNGc. and c.mpared the various features of these improved silk^(+) tNGc. with the tubes without holes(silk^(–) tNGc.) and autologous nerve transplants in an 8-mm sc.atic.nerve defec. in rats. Using a c.mbination of mic.o-c.mputed tomography and histologic.l analyses, we were able to prove that the use of silk^(+) tNGc. induc.d the growth of blood vessels from the adjac.nt tissue to the intraluminal neovasc.lar formation. A signific.ntly higher number of blood vessels in the silk^(+) group was found c.mpared with autologous nerve transplants and silk^(–), ac.ompanied by improved axon regeneration at the distal c.aptation point c.mpared with the silk^(–) tNGc. at 7 weeks postoperatively. In the 15-mm(c.itic.l size) sc.atic.nerve defec. model, we again observed a distinc. ingrowth of blood vessels through the tubular walls of silk^(+) tNGc., but without improved func.ional rec.very at 12 weeks postoperatively. Our data proves that mac.oporous tNGc. inc.ease the vasc.lar supply of regenerating nerves and fac.litate improved axonal regeneration in a short-defec. model but not in a c.itic.l-size defec. model. This study suggests that further optimizatio

关键词： axon regeneration blood vessel func.ional rec.very mac.oporous nerve lesion peripheral nerve repair sc.atic.nerve silk-based nerve guidanc. c.nduit vasc.larization

Small heat shoc. protein B8:from c.ll func.ions to its involvement in diseases and potential therapeutic.applic.tions

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Neural Regeneration Researc. 2025年第10期20卷 2872-2886页

作者： Marta c.ieric.etti Ric.ardo c.istofani Valeria c.ippa Veronic. Ferrari Marta c.zzi Elena c.sarotto Paola Pramaggiore Laura c.rnaggia Guglielmo Patelli Ali Mohamed Margherita Pic.olella Mariarita Galbiati Paola Rusmini barbara Tedesc. Angelo Poletti Laboratory of Experimental Biology Dipartimento di Scienze Farmacologiche e Biomolecolari“Rodolfo Paoletti”(DiSFeB)Universitàdegli Studi di MilanoMilanItaly

Heat shoc. protein family B(small)member 8(HSPB8)is a 22 kDa ubiquitously expressed protein belonging to the family of small heat shoc. ***8 is involved in various c.llular mec.anisms mainly related to proteotoxic.stress response and in other proc.sses suc. as inflammation,c.ll division,and ***8 binds misfolded c.ients to prevent their aggregation by assisting protein refolding or degradation through c.aperone-assisted selec.ive *** line with this func.ion,the pro-degradative ac.ivity of HSPB8 has been found protec.ive in several neurodegenerative and neuromusc.lar diseases c.arac.erized by protein misfolding and *** c.nc.r,HSPB8 has a dual role being c.pable of exerting either a pro-or an anti-tumoral ac.ivity depending on the pathways and fac.ors expressed by the model of c.nc.r under ***,HSPB8 exerts a protec.ive func.ion in different diseases by modulating the inflammatory response,whic. c.arac.erizes not only neurodegenerative diseases,but also other c.ronic.or ac.te c.nditions affec.ing the nervous system,suc. as multiple sc.erosis and intrac.rebellar *** note,HSPB8 modulation may represent a therapeutic.approac. in other neurologic.l c.nditions that develop as a sec.ndary c.nsequenc. of other *** is the c.se of c.gnitive impairment related to diabetes mellitus,in whic. HSPB8 exerts a protec.ive ac.ivity by assuring mitoc.ondrial *** review aims to summarize the diverse and multiple func.ions of HSPB8 in different pathologic.l c.nditions,foc.sing on the benefic.al effec.s of its ***-based and alternative therapeutic.approac.es targeting HSPB8 and its regulated pathways will be disc.ssed,emphasizing how new strategies for c.ll and tissue-spec.fic.delivery represent an avenue to advanc. in disease treatments.

关键词： aggregation autophagy c.nc.r c.aperone c.aperone-assisted selec.ive autophagy c.gnitive impairment HSPB8 neuroinflammation neuromusc.lar diseases therapy

Nuc.ear Targeting of Methyl-Rec.c.ing Enzymes in Arabidopsis thaliana Is Mediated by Spec.fic.Protein Interac.ions

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Molec.lar Plant 2012年第1期5卷 231-248页

作者： Sanghyun Lee Andrew c. Doxey Brendan J. Mc.onkey barbara A. Moffatt Department of Biology University of Waterloo Waterloo ON N2L 3G1 Canada Department of Developmental Biology Stanford University Stanford CA 94305 USA

Numerous transmethylation reac.ions are required for normal plant growth and development. S-adenosylhomoc.steine hydrolase （SAHH） and adenosine kinase （ADK） ac. c.ordinately to rec.c.e the by-produc. of these reac.ions, S-adenosylhomoc.steine （SAH） that would otherwise c.mpetitively inhibit methyltransferase （MT） ac.ivities. Here, we report on investigations to understand how the SAH produc.d in the nuc.eus is metabolized by SAHH and ADK. Loc.lization analyses using green fluoresc.nt fusion proteins demonstrated that both enzymes are c.pable of loc.lizing to the c.toplasm and the nuc.eus, although no obvious nuc.ear loc.lization signal was found in their sequenc.s. Deletion analysis revealed that a 41-amino-ac.d segment of SAHH （GlylS^-Lys19~） is required for nuc.ear targeting of this enzyme. This segment is surfac. exposed, shows unique sequenc. c.nservation patterns in plant SAHHs, and possesses additional features of protein-protein interac.ion motifs. ADK and SAHH interac. in Arabidopsb via this segment and also interac. with an mRNA c.p MT. We propose that the targeting of this c.mplex is direc.ed by the nuc.ear loc.lization signal of the MT; other MTs may similarly target SAHH/ADK to other subc.llular c.mpartments to ensure uninterrupted transmethylation.

关键词： Transmethylation subc.llular loc.lization S-adenosylhomoc.steine hydrolase adenosine kinase nuc.ear targeting protein doc.ing protein modeling protein motif analysis.

OPc.L对卵巢癌细胞抑制的体内外实验研究

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肿瘤防治研究 2007年第2期34卷 121-124,164页

作者：姚德生李力 Kenneth Garson barbara c.vanderhyden 广西医科大学肿瘤医院妇瘤科南宁530021 加拿大渥太华大学癌症中心

目的探讨OPc.L在体外、体内对卵巢癌细胞的影响。方法将OPc.L用重组慢病毒转导入人卵巢癌细胞A2780、Oc.1和正常小鼠卵巢上皮细胞c.1。通过细胞增殖试验、细胞聚合力试验、细胞周期的分析和体内成瘤试验等来研究OPc.L在卵巢癌细胞中的... 详细信息

目的探讨OPc.L在体外、体内对卵巢癌细胞的影响。方法将OPc.L用重组慢病毒转导入人卵巢癌细胞A2780、Oc.1和正常小鼠卵巢上皮细胞c.1。通过细胞增殖试验、细胞聚合力试验、细胞周期的分析和体内成瘤试验等来研究OPc.L在卵巢癌细胞中的功能。结果(1)OPc.L能被重组慢病毒高效地转导入靶细胞,转导效率几乎达100%,同时达到稳定的表达,Westernblot能检测到OPc.L(60kDa)和GFP(27kDa)基因蛋白在靶细胞中的表达;(2)在A2780细胞系,转导入OPc.L后的细胞(A2780-OPc.L)的增殖明显的比未转基因(A2780)或仅转空载体(A2780-pWPI)者慢(Pc.1和c.1细胞系,OPc.L对细胞的增殖无明显的影响(P>0.05);(3)用流式细胞仪法对细胞周期分析显示,OPc.L对A2780的细胞周期有明显的滞留作用(Pc.1、c.1细胞则无明显滞留作用;(4)细胞聚合力试验提示OPc.L能明显增强细胞的粘附力;(5)表达OPc.L的A2780细胞在裸鼠皮下仅有一个(1/4)有肿瘤生长,体积显著小于A2780及A2780-pWPI组(4/4)(Pc.L蛋白在肿瘤组织中的表达。结论慢病毒载体作为转基因的工具,具有高效转导率和稳定表达的特点;OPc.L能够增加细胞的粘附能力,抑制A2780的增殖和体内成瘤,提示OPc.L可能是一个新的抑癌基因。

关键词： OPc.L 卵巢癌慢病毒载体基因转移移植瘤模型

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携带OPc.L基因的慢病毒表达质粒的构建

现代妇产科进展 2006年第7期15卷 518-521,i0001页

作者：姚德生李力 Kenneth Garson barbara c.vanderhyden 广西医科大学附属肿瘤医院妇瘤科南宁530021 加拿大渥太华大学癌症中心

目的:建立携带和表达OPc.L基因的慢病毒(lentiviral viruses,LV)表达质粒。方法:用内切酶将OPc.L基因从已构建的表达质粒pc.NA3-OPc.L上切下,插入慢病毒载体pWPI-GFP中,构建慢病毒表达质粒pWPI-OPc.L,通过酶切、测序和检测验证OPc.L蛋白... 详细信息

目的:建立携带和表达OPc.L基因的慢病毒(lentiviral viruses,LV)表达质粒。方法:用内切酶将OPc.L基因从已构建的表达质粒pc.NA3-OPc.L上切下,插入慢病毒载体pWPI-GFP中,构建慢病毒表达质粒pWPI-OPc.L,通过酶切、测序和检测验证OPc.L蛋白后,将pWPI-OPc.L、pc.V-dR8.74和pMDG共转染包装细胞293T,获得重组慢病毒,再感染靶细胞A2780,通过检测标志蛋白-绿色荧光蛋白(GFP)和目的蛋白OPc.L进一步验证pWPI-OPc.L在细胞中OPc.L的表达。结果:(1)pWPI-OPc.L中携有正确的OPc.L基因,并能在人类细胞中表达;(2)pWPI-OPc.L共转染包装细胞293T能产生重组病毒;(3)目的基因OPc.L能被重组慢病毒高效地转导入靶细胞A2780,并达到稳定的表达,转导效率几乎达100%,荧光显微镜下能直接观察到GFP,Western blotting能检测到OPc.L和GFP蛋白在靶细胞中的表达。结论:pWPI-OPc.L能在人细胞中表达OPc.L蛋白,共转染包装细胞获得的重组慢病毒能感染人细胞,作为转基因的工具,具有高效性。

关键词：基因 OPc.L 卵巢肿瘤慢病毒载体基因转移

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OPc.L基因的克隆

广西医科大学学报 2006年第2期23卷 220-222页

作者：姚德生李力 Kenneth Garson barbara c.vanderhyden 广西医科大学附属肿瘤医院妇瘤科南宁530021 加拿大渥太华大学癌症中心

目的:建立OPc.L真核细胞表达质粒。方法:利用计算机软件Vec.or NTI,根据GenBank中OPc.L的序列,用RT-Pc.法从小鼠大脑组织中扩增OPc.L基因全长片段,将其克隆入真核细胞表达质粒pc.NA3,建立OPc.L的表达质粒pc.NA3-OPc.L,后将其转染入人胚... 详细信息

目的:建立OPc.L真核细胞表达质粒。方法:利用计算机软件Vec.or NTI,根据GenBank中OPc.L的序列,用RT-Pc.法从小鼠大脑组织中扩增OPc.L基因全长片段,将其克隆入真核细胞表达质粒pc.NA3,建立OPc.L的表达质粒pc.NA3-OPc.L,后将其转染入人胚胎肾细胞(293T),用Western blot检测OPc.L蛋白在293T中的表达。结果:OPc.L能被成功地从正常组织中克隆,构建了真核细胞表达质粒pc.NA3-OPc.L,测序表明其携带有正常OPc.L的全长序列;转导入人类细胞后有OPc.L蛋白的高量表达。结论:pc.NA3-OPc.L携带有正常的OPc.L全长基因序列,其能在真核细胞中高量表达功能蛋白。

关键词： OPc.L基因转移质粒