摘要
目的:检测人胃癌细胞系中FHIT基因mRNA的表达状况及构建pcDNA3.1-FHIT真核表达载体。方法:RT-PCR法检测三种不同类型人胃癌细胞系中FHIT基因mRNA的表达,构建真核表达质粒pcDNA3.1-FHIT,通过酶切法、PCR扩增法和DNA序列分析鉴定重组质粒后,用脂质体转染至FHIT基因mRNA阴性表达人胃癌细胞系MKN-45,经G418筛选后RT-PCR鉴定。结果:FHIT基因在人胃癌细胞系BGC-823中呈阳性表达,在MGC-803、MKN-45细胞系中呈阴性表达。FHIT基因cDNA正确克隆到真核细胞表达载体pcDNA3.1中,并成功转染FHIT基因mRNA阴性表达人胃癌细胞系MKN-45。结论:FHIT基因在不同类型人胃癌细胞系中表达各异。成功构建pcDNA3.1-FHIT,并转染到FHIT基因mRNA阴性表达人胃癌细胞系MKN-45,使其FHIT基因阳性表达。
Objective：To investigate the expression of FHIT in gastric cancer cell lines and construct recombinant plasmid pcDNA3.1-FHIT and transfect it.Methods：Investigate the expression of FHIT in three gastric cancer cell lines by RT-PCR,clone human FHIT cDNA from a gastric cancer cell line with plasmid pcDNA3.1,to construct eukaryotic expression carrier of recombinant pcDNA3. 1-FHIT,which was transfected into human gastric cancer cell line MKN-45 by liposome after identification by PCR,restriction enzyme digesting and DNA sequencing.The stomach cancer cells were selected by G418,identified by RT-PCR.Results：There was a high expression of FHIT in BGC-823,no expression in MGC-803 and MKN-45.The recombinant plasmid pcDNA3.1 was successfully constructed and transfected intoMKN-45.Conclusions：The different expression levels of FHIT in three stomach cancer cells were conformed and the pcDNA3.1-FHIT has been constructed and transfected successfully,which makes FHIT highly expressed in FHIT-regenerating cell line MKN-45.
出处
《现代生物医学进展》
CAS
2008年第8期1419-1421,1448,共4页
Progress in Modern Biomedicine
基金
右江民族医学院科研基金资助。
关键词
FHIT基因
胃癌
基因表达
构建
转染
FHIT gene
stomach cancer
gene expression
construction
transfection
