生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2013年
2期
193-196
,共4页
罗国兰%王洪涛%伍飞马%李海量%张晓清%薛萌%王健%李山虎%赵亚丽%刘萱%周建光
囉國蘭%王洪濤%伍飛馬%李海量%張曉清%薛萌%王健%李山虎%趙亞麗%劉萱%週建光
라국란%왕홍도%오비마%리해량%장효청%설맹%왕건%리산호%조아려%류훤%주건광
4E-BP1%4E-BP1-4A%慢病毒载体%胃癌%细胞生长
4E-BP1%4E-BP1-4A%慢病毒載體%胃癌%細胞生長
4E-BP1%4E-BP1-4A%만병독재체%위암%세포생장
4E-BP1%4E-BP1-4A%lentivirus vector%gastric cancer%cell growth
目的:构建4E-BP1及其 T37A、T46A、S65A、T70A 突变体4E-BP1-4A 基表达的重组慢病毒载体,研究其对胃癌 HGC27细胞生长的影响.方法:PCR 扩增4E-BP1基及其突变体4E-BP1-4A 基并克隆到 pCDH 载体,构建成 pCDH-4E-BP1、pCDH-4E-BP1-4A,将其与包装载体共转染293T 细胞,包装成 Lenti-4E-BP1及 Lenti-4E-BP1-4A重组慢病毒载体,将此慢病毒感染胃癌 HGC27细胞,Western 印迹鉴定病毒载体介导的4E-BP1、4E-BP1-4A 蛋白的表达,MTT、克隆形成和软琼脂方法研究过量表达4E-BP1、4E-BP1-4A 对胃癌 HGC27细胞生长的影响.结果:包装成 Lenti-4E-BP1及 Lenti-4E-BP1-4A 重组慢病毒载体,并将此慢病毒载体感染胃癌 HGC27细胞;MTT、克隆形成、软琼脂实验表明过量表达4E-BP1可抑制胃癌 HGC27细胞的生长,过量表达4E-BP1-4A 时抑制效果更明显.结论:构建了4E-BP1、4E-BP1-4A 的重组慢病毒表达载体,在胃癌 HGC27细胞中过量表达4E-BP1可抑制细胞生长,过量表达4E-BP1-4A 的抑制效果更明显.
目的:構建4E-BP1及其 T37A、T46A、S65A、T70A 突變體4E-BP1-4A 基錶達的重組慢病毒載體,研究其對胃癌 HGC27細胞生長的影響.方法:PCR 擴增4E-BP1基及其突變體4E-BP1-4A 基併剋隆到 pCDH 載體,構建成 pCDH-4E-BP1、pCDH-4E-BP1-4A,將其與包裝載體共轉染293T 細胞,包裝成 Lenti-4E-BP1及 Lenti-4E-BP1-4A重組慢病毒載體,將此慢病毒感染胃癌 HGC27細胞,Western 印跡鑒定病毒載體介導的4E-BP1、4E-BP1-4A 蛋白的錶達,MTT、剋隆形成和軟瓊脂方法研究過量錶達4E-BP1、4E-BP1-4A 對胃癌 HGC27細胞生長的影響.結果:包裝成 Lenti-4E-BP1及 Lenti-4E-BP1-4A 重組慢病毒載體,併將此慢病毒載體感染胃癌 HGC27細胞;MTT、剋隆形成、軟瓊脂實驗錶明過量錶達4E-BP1可抑製胃癌 HGC27細胞的生長,過量錶達4E-BP1-4A 時抑製效果更明顯.結論:構建瞭4E-BP1、4E-BP1-4A 的重組慢病毒錶達載體,在胃癌 HGC27細胞中過量錶達4E-BP1可抑製細胞生長,過量錶達4E-BP1-4A 的抑製效果更明顯.
목적:구건4E-BP1급기 T37A、T46A、S65A、T70A 돌변체4E-BP1-4A 기표체적중조만병독재체,연구기대위암 HGC27세포생장적영향.방법:PCR 확증4E-BP1기급기돌변체4E-BP1-4A 기병극륭도 pCDH 재체,구건성 pCDH-4E-BP1、pCDH-4E-BP1-4A,장기여포장재체공전염293T 세포,포장성 Lenti-4E-BP1급 Lenti-4E-BP1-4A중조만병독재체,장차만병독감염위암 HGC27세포,Western 인적감정병독재체개도적4E-BP1、4E-BP1-4A 단백적표체,MTT、극륭형성화연경지방법연구과량표체4E-BP1、4E-BP1-4A 대위암 HGC27세포생장적영향.결과:포장성 Lenti-4E-BP1급 Lenti-4E-BP1-4A 중조만병독재체,병장차만병독재체감염위암 HGC27세포;MTT、극륭형성、연경지실험표명과량표체4E-BP1가억제위암 HGC27세포적생장,과량표체4E-BP1-4A 시억제효과경명현.결론:구건료4E-BP1、4E-BP1-4A 적중조만병독표체재체,재위암 HGC27세포중과량표체4E-BP1가억제세포생장,과량표체4E-BP1-4A 적억제효과경명현.
Objective: To construct and identify combinant lentiviral vectors for overexpression of 4E-BP1 and the T37A, T46A, S65A, T70A mutant of 4E-BP1-4A, and to study their effects on the grown of the gastric can?cer cell HGC27. Methods: 4E-BP1 and 4E-BP1-4A genes were amplified using PCR and cloned into the pCDH vector. The resulting plasmid pCDH-4E-BP1 and pCDH-4E-BP1-4A with the packing carrier mix were co-trans?fected into 293T cells to obtain packaged lentivirusparticles, and then the packaged lentivirusparticles were used to infect HGC27 cells. Western blot was performed to identify lentivirus-mediated expression of 4E-BP1 and 4E-BP1-4A in HGC27 cells. MTT and colony-formation assays were performed to study the effect of overexpres?sion of 4E-BP1 and 4E-BP1-4A on the grown of HGC27 cells. Results: The recombinant lentivirus expressed 4E-BP1 and 4E-BP1-4A efficiently in HGC27 cells. The MTT and colony-formation assays indicated that overex?pression of 4E-BP1 and 4E-BP1-4A inhibited the growth of HGC27 cells. Conclusion: Recombinant lentiviruses overexpressing 4E-BP1 and 4E-BP1-4A were successfully constructed. Overexpression of 4E-BP1 and 4E-BP1-4A suppressed the growth of HGC27 cells.
