Construction of Formin-like 2 specific small hairpin RNA expressing plasmid and its effect on proliferation in SW620
- VernacularTitle:FMNL2 shRNA表达质粒的构建及其对SW620增殖活性的影响
- Author:
Xiling ZHU
;
Li LIANG
;
Yanqing DING
- Publication Type:Journal Article
- Keywords:
Short hairpin RNA;
Formin-like 2;
Proliferation
- From:
Clinical Medicine of China
2008;24(6):513-517
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the specific small hairpin RNA(shRNA)plasmids for Formin-like 2(FMNL2)gene,and to observe its effect on proliferation of SW620.Methods Plasmids FMNL21 shRNA and plasmids FMNL22 shRNA complimentary to the sequence responsible for the function domain of human FMNL2 were constructed and transfected into SW620,respectively.The inhibition rates of FMNL2 expression were detected by re-al-time PCR after transfection with PGenesil-FMNL21 and PGenesil-FMNL22,respectively.The expression of FMNL2 was detected by real-time PCR at 12,24,48,72 and 96h after transfection.At 48 h and 72 h after transfection,the proliferation of SW620 was detected using MTT assay.Resuits The transfection rate in SW620 was(60.8±2.8)%for PGenesil-FMNL21,(58.7±2.9)%for PGenesil-FMNL22,and(62.6±1.7)%for PGenesil-HK(P＞0.05).The inhibition rate of FMNL2 expression after transfection with PGenesil-FMNL21 was 77.1%,which was the highest.The inhibition rates of FMNL2 expression were 13.5%,57.3%,80.3%,62.4%,and 33.2%at 12,24,48,72and 96h after transfection respectively.The proliferat capability of SW620 was obviously lower than that of control cells(P＜0.05).Conclusion Both plasmids FMNL21 shRNA and plasmids FMNL22 shRNA can inhibit the expression of FMNL2,of which Pgenesil-FMNL21 execs the greatest role.FMNL2 could be related to the proliferation of SW620.
