CAJ | 학술논문

背景:通过测序进行序列分析是发现基因突变的直接手段,转化生长因子β1(TGF-β1)作为细胞最主要的生长调控因子,参与多项生理和病理过程.该基因的突变对于所在细胞的生物学行为改变可能具有重要意义.目的:分析人成骨肉瘤细胞系中TGF-β1成熟肽基因序列,探讨TGF-β1基因突变存在的可能性及其机制,为肿瘤及骨软骨损伤的基因治疗提供新思路.设计:非随机非对照实验研究.地点和对象:实验在第四军医大学唐都医院解放军骨科中心骨肿瘤研究所完成,对象为人成骨肉瘤细胞系SOSP-9607(本室建立).干预:提取人成骨肉瘤细胞系SOSP-9607中的总RNA,参照Genebank数据库中的已知序列设计合成引物,采用RT-PCR法扩增TGF-β1成熟肽基因,克隆入质粒pcDNA 3.0进行序列测定和分析.主要观察指标:将多次测序所得的TGF-β1成熟肽基因序列与Genebank数据库中的已知序列进行比较,分析恒定差异.结果:发现人成骨肉瘤细胞系SOSP-9607中TGF-β1基因存在两处点突变,其中第1 788位突变引起相应多肽的结构改变.结论:人成骨肉瘤细胞系SOSP-9607中存在TGF-β1基因突变,其可能在病理机制中起重要作用,具有强功能活性突变子的获得对于骨软骨损伤的修复有重要意义.
배경:통과측서진행서렬분석시발현기인돌변적직접수단,전화생장인자β1(TGF-β1)작위세포최주요적생장조공인자,삼여다항생리화병리과정.해기인적돌변대우소재세포적생물학행위개변가능구유중요의의.목적:분석인성골육류세포계중TGF-β1성숙태기인서렬,탐토TGF-β1기인돌변존재적가능성급기궤제,위종류급골연골손상적기인치료제공신사로.설계:비수궤비대조실험연구.지점화대상:실험재제사군의대학당도의원해방군골과중심골종류연구소완성,대상위인성골육류세포계SOSP-9607(본실건립).간예:제취인성골육류세포계SOSP-9607중적총RNA,삼조Genebank수거고중적이지서렬설계합성인물,채용RT-PCR법확증TGF-β1성숙태기인,극륭입질립pcDNA 3.0진행서렬측정화분석.주요관찰지표:장다차측서소득적TGF-β1성숙태기인서렬여Genebank수거고중적이지서렬진행비교,분석항정차이.결과:발현인성골육류세포계SOSP-9607중TGF-β1기인존재량처점돌변,기중제1 788위돌변인기상응다태적결구개변.결론:인성골육류세포계SOSP-9607중존재TGF-β1기인돌변,기가능재병리궤제중기중요작용,구유강공능활성돌변자적획득대우골연골손상적수복유중요의의.
BACKGROUND: It is a direct means in the detection of gene mutation to analyze the gene sequences. Transforning growth factor β1 (TGF-β1), as the most important celhdar growth control and regulation factor, participates in many physiological and pathological processes. The mutation of this gene might have significances in the changes of cellular biological behaviors.OBJECTIVE: To analyze the gene sequence of mature peptide of TGF-β1 in human osteogenic sarcoma in order to discuss the possibility and mechanism of the existence of TGF-β1 mutation to provide a new approach to gene therapy of tumors and bone and cartilage injuries.DESIGN: A non-randomized, non-controlled study was conducted.SETTING and PARTICIPANTS: The experiment was completed in the Center of Orthopaedic Surgery, Orthopaedic Oncology Institute of PLA,Tangdu Hospital, Fourth Military Medical University. The subjects were human osteogenic sarcoma serials SOSP-9607 established by our department.INTERVENTIONS: Total RNA in SOSP-9607 was extracted. Primer was designed and synthesized according to the known sequence from Genebank database. RT-PCR was introduced to amplify and cloned into plasmid pcDNA 3.0 for sequence assay and analysis.MAIN OUTCOME MEASURES: Mature peptide gene TGF-31 obtained from many times of sequencing was compared with the known sequence in Genebank database and the constant difference was analyzed.RESULTS: There were two point mutations were discovered in TGF-β1 of SOSP-9607, in which the mutation in the 1788th would induce structure changes in corresponding polypeptide.CONCLUSION: There were gene mutations of TGF-1 in SOSP-9607,which might play an important role in pathological mechanism. The gain of muton with strong functional activity is very important for the repair of bone and cartilage injuries.