中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
6期
816-818
,共3页
李兆明%董硕%刘韦成%蔡少鑫%罗学来%李小兰%陶德定%严群%龚建平%胡俊波
李兆明%董碩%劉韋成%蔡少鑫%囉學來%李小蘭%陶德定%嚴群%龔建平%鬍俊波
리조명%동석%류위성%채소흠%라학래%리소란%도덕정%엄군%공건평%호준파
hSav1%细胞增殖%人胚肾细胞
hSav1%細胞增殖%人胚腎細胞
hSav1%세포증식%인배신세포
hSav1%Cell proliferation%Human embryonic kidney cell
目的 观察高表达人Salvadorl(hSav1)蛋白对人胚肾293(HEK293)细胞增殖的影响.方法 构建含有绿色荧光蛋白(GFP)的hSav1质粒GFP-N1-hSav1;将构建好的GFP-N1-hSav1质粒在脂质体Lipofectamine 2000的介导下转染HEK293细胞,荧光显微镜下观察质粒转染效率;分别在转染后12、24、36、48 h通过噻唑蓝(MTT)比色法计算细胞增殖抑制率,抑制率=[(对照-空白)-(给药-空白)]/(对照-空白)×100%;转染后36 h加入5-溴-2'-脱氧尿苷(BrdU),通过其掺入比率来显示hSav1对细胞增殖的影响.结果 质粒GFP-N1-hSav1构建成功,转染效率为70%-80%左右,蛋白hSav1明显高表达;MTT结果显示转染质粒后12、24、36、48 h,高表达蛋白hSav1组的细胞增殖抑制率分别为2%、5%、15%、23%,而阴性对照分别为2%、3%、2%、2%;BrdU结果显示,在HEK293细胞中高表达蛋白hSav1后细胞增殖抑制率为(27.3±3.8)%,阴性对照组的细胞增殖抑制率为(12.9±5.3)%,差异有统计学意义(P<0.05).结论 在人胚肾HEK293中高表达蛋白hSav1可明显抑制细胞增殖能力.
目的 觀察高錶達人Salvadorl(hSav1)蛋白對人胚腎293(HEK293)細胞增殖的影響.方法 構建含有綠色熒光蛋白(GFP)的hSav1質粒GFP-N1-hSav1;將構建好的GFP-N1-hSav1質粒在脂質體Lipofectamine 2000的介導下轉染HEK293細胞,熒光顯微鏡下觀察質粒轉染效率;分彆在轉染後12、24、36、48 h通過噻唑藍(MTT)比色法計算細胞增殖抑製率,抑製率=[(對照-空白)-(給藥-空白)]/(對照-空白)×100%;轉染後36 h加入5-溴-2'-脫氧尿苷(BrdU),通過其摻入比率來顯示hSav1對細胞增殖的影響.結果 質粒GFP-N1-hSav1構建成功,轉染效率為70%-80%左右,蛋白hSav1明顯高錶達;MTT結果顯示轉染質粒後12、24、36、48 h,高錶達蛋白hSav1組的細胞增殖抑製率分彆為2%、5%、15%、23%,而陰性對照分彆為2%、3%、2%、2%;BrdU結果顯示,在HEK293細胞中高錶達蛋白hSav1後細胞增殖抑製率為(27.3±3.8)%,陰性對照組的細胞增殖抑製率為(12.9±5.3)%,差異有統計學意義(P<0.05).結論 在人胚腎HEK293中高錶達蛋白hSav1可明顯抑製細胞增殖能力.
목적 관찰고표체인Salvadorl(hSav1)단백대인배신293(HEK293)세포증식적영향.방법 구건함유록색형광단백(GFP)적hSav1질립GFP-N1-hSav1;장구건호적GFP-N1-hSav1질립재지질체Lipofectamine 2000적개도하전염HEK293세포,형광현미경하관찰질립전염효솔;분별재전염후12、24、36、48 h통과새서람(MTT)비색법계산세포증식억제솔,억제솔=[(대조-공백)-(급약-공백)]/(대조-공백)×100%;전염후36 h가입5-추-2'-탈양뇨감(BrdU),통과기참입비솔래현시hSav1대세포증식적영향.결과 질립GFP-N1-hSav1구건성공,전염효솔위70%-80%좌우,단백hSav1명현고표체;MTT결과현시전염질립후12、24、36、48 h,고표체단백hSav1조적세포증식억제솔분별위2%、5%、15%、23%,이음성대조분별위2%、3%、2%、2%;BrdU결과현시,재HEK293세포중고표체단백hSav1후세포증식억제솔위(27.3±3.8)%,음성대조조적세포증식억제솔위(12.9±5.3)%,차이유통계학의의(P<0.05).결론 재인배신HEK293중고표체단백hSav1가명현억제세포증식능력.
Objective To elucidate the effects of human Salvador 1 (hSav1 ) on cell proliferation of human embryonic kidney cell line HEK293. Methods The plasmid CFP-N1-hSav1 was constructed and transfected into HEK293 cells with lipofectamine 2000. The transfection efficiency was detected by fluorescent microscopy. The effects of hSav1 on cell proliferation were measured by MTT and BrdU incorporation. Results The transfection efficiency was about 70% -80%. The MTT results showed that the inhibition rate of cell proliferation in transfected group at the 12th, 24th, 36th, and 48th h was 2% , 5% , 15% , and 23% , respectively;while that in the control group was 2% , 3% , 2% , and 2% respectively. The BrdU incorporation revealed that the BrdU incorporation rate in transfected group (12. 9 ±5. 3)% was significantly lower than in control group (27.3±3.8)% (P<0.05). Conclusion hSav1 is a newly identified protein that can cause cell proliferation inhibition in HEK293 cells.
