CAJ | 학술논문

目的构建pEGFP-金属调理素-1(MPS-1)真核表达载体,体外转染至SGC-7901胃癌细胞,观察MPS-1转染后MPS-1蛋白表达,并检测SGC-7901细胞侵袭能力的变化.方法聚合酶链反应(PCR)扩增MPS-1全长255 bp的编码序列,将PCR扩增产物连接入pEGFP-C1载体,经测序确认序列无误后成功构建pEGFP-MPS-1真核表达载体.采用Lipofectamine 2000转染SGC-7901胃癌细胞株,用浓度为2.0 g/L的G418筛选2周后得到外源性MPS-1稳定表达的SGC-7901细胞并扩增.Western blot法检测MPS-1的表达,Transwell实验检测pEGFP-MPS-1过表达后对胃癌细胞侵袭能力的影响.结果得到外源性MPS-1稳定表达的胃癌细胞株,Western blot显示外源性MPS-1在胃癌SGC-7901细胞中表达成功,而且,pEGFP-MPS-1过表达后胃癌细胞的Transwell实验显示pEGFPMPS-1过表达细胞组相对于空载组和空白组的侵袭指数分别为2.10 ±0.25比1.00 ±0.00和2.10±0.25比0.85±0.22(P＜0.05),提示MPS-1稳定表达后胃癌细胞的侵袭能力增加.结论 pEGFPMPS-1的稳定表达增加了胃癌细胞SGC-7901的侵袭能力.
목적 구건pEGFP-금속조리소-1(MPS-1)진핵표체재체,체외전염지SGC-7901위암세포,관찰MPS-1전염후MPS-1단백표체,병검측SGC-7901세포침습능력적변화.방법 취합매련반응(PCR)확증MPS-1전장255 bp적편마서렬,장PCR확증산물련접입pEGFP-C1재체,경측서학인서렬무오후성공구건pEGFP-MPS-1진핵표체재체.채용Lipofectamine 2000전염SGC-7901위암세포주,용농도위2.0 g/L적G418사선2주후득도외원성MPS-1은정표체적SGC-7901세포병확증.Western blot법검측MPS-1적표체,Transwell실험검측pEGFP-MPS-1과표체후대위암세포침습능력적영향.결과 득도외원성MPS-1은정표체적위암세포주,Western blot현시외원성MPS-1재위암SGC-7901세포중표체성공,이차,pEGFP-MPS-1과표체후위암세포적Transwell실험현시pEGFPMPS-1과표체세포조상대우공재조화공백조적침습지수분별위2.10 ±0.25비1.00 ±0.00화2.10±0.25비0.85±0.22(P＜0.05),제시MPS-1은정표체후위암세포적침습능력증가.결론 pEGFPMPS-1적은정표체증가료위암세포SGC-7901적침습능력.
Objective To construct the expression vector of pEGFP-metallopanstimulin-1 (MPS-1),observe the ectopic MPS-1 protein expression in SGC-7901 cells,and examine the change in invasive capability of SGC-7901 cells.Methods The coding sequence of MPS-1 255 bp was obtained by polymerase chain reaction (PCR) amplification,then the PCR product was inserted into the pEGFP-C1 vector.After confirming the DNA sequence the MPS-1 gene eukaryotic expression vector was obtained.The pEGFP-MPS-1 expression vector was transfected into SGC-7901 gastric cancer cells by Lipofectamine 2000,G418 2.0 g/L was used to screen the cells and stable cells were obtained after two weeks.The expression of pEGFP-MPS-1 was detected by using Western blotting.The changes in invasive capability of SGC-7901 cells was examined by using Transwell.Results The recombinant vector was highly expressed in SGC-7901 cells.Over-expression of pEGFP-MPS-1 enhanced the invasive ability of SGC-7901 cells (P ＜ 0.05).The invasion index in pEGFP-MPS-1 overexpression group was 2.10 ± 0.25,significantly higher than in vector group (1.00 ± 0.00) and blank group (0.85 ± 0.22) (P ＜ 0.05).Conclusion The stable expression of pEGFP-MPS-1 enhanced the invasive capability of SGC-7901 cells.