中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2009年
4期
237-241
,共5页
贺钰磊%曹励之%杨明华%赵明一%俞燕%许望琼
賀鈺磊%曹勵之%楊明華%趙明一%俞燕%許望瓊
하옥뢰%조려지%양명화%조명일%유연%허망경
WASP家族富含脯氨酸同源蛋白1%基质金属蛋白酶%K562细胞系%RNA干扰%白血病浸润
WASP傢族富含脯氨痠同源蛋白1%基質金屬蛋白酶%K562細胞繫%RNA榦擾%白血病浸潤
WASP가족부함포안산동원단백1%기질금속단백매%K562세포계%RNA간우%백혈병침윤
WAVE1%MMP-2%K562 cell line%RNA interference%Invasion
目的 研究WASP家族富含脯氨酸同源蛋白1(WAVE1)对K562细胞侵袭的影响及其机制.方法 免疫荧光观察WAVE1与基质金属蛋白-2(MMP-2)在细胞中的分布.利用PeDNA3.1WAVE1重组真核表达质粒转染K562细胞,将WAVE1基因特异性小片段干扰RNA(WAVEI siRNA)转染K562细胞,Transwell法检测细胞侵袭能力;实时PCR和Western blot检测转染前后WAVE1及MMP-2的表达.结果 ①WAVEI与MMP-2主要表达于K562细胞的细胞膜上且两者有共定位.②与对照组K562细胞相比,转染pcDNA3.1-WAVE1 24 h及48 h后K562细胞MMP-2 mRNA表达分别增加295%和198%,蛋白表达水平分别增加80%和23%;转染特异性WAVE1 siRNA 24 h及48 h后K562细胞MMP-2 mRNA表达分别下降81%和28%,蛋白表达分别下调36%和53%.③与对照组K562细胞相比转染pcDNA3.1-WAVE1后细胞侵袭能力增强,而干扰WAVE1表达后K562细胞侵袭能力减弱.结论 WAVE1与MMP-2在K562细胞可能具有协同作用;WAVE1参与了K562细胞的侵袭转移过程,其机制可能与调控MMP-2的表达相关.
目的 研究WASP傢族富含脯氨痠同源蛋白1(WAVE1)對K562細胞侵襲的影響及其機製.方法 免疫熒光觀察WAVE1與基質金屬蛋白-2(MMP-2)在細胞中的分佈.利用PeDNA3.1WAVE1重組真覈錶達質粒轉染K562細胞,將WAVE1基因特異性小片段榦擾RNA(WAVEI siRNA)轉染K562細胞,Transwell法檢測細胞侵襲能力;實時PCR和Western blot檢測轉染前後WAVE1及MMP-2的錶達.結果 ①WAVEI與MMP-2主要錶達于K562細胞的細胞膜上且兩者有共定位.②與對照組K562細胞相比,轉染pcDNA3.1-WAVE1 24 h及48 h後K562細胞MMP-2 mRNA錶達分彆增加295%和198%,蛋白錶達水平分彆增加80%和23%;轉染特異性WAVE1 siRNA 24 h及48 h後K562細胞MMP-2 mRNA錶達分彆下降81%和28%,蛋白錶達分彆下調36%和53%.③與對照組K562細胞相比轉染pcDNA3.1-WAVE1後細胞侵襲能力增彊,而榦擾WAVE1錶達後K562細胞侵襲能力減弱.結論 WAVE1與MMP-2在K562細胞可能具有協同作用;WAVE1參與瞭K562細胞的侵襲轉移過程,其機製可能與調控MMP-2的錶達相關.
목적 연구WASP가족부함포안산동원단백1(WAVE1)대K562세포침습적영향급기궤제.방법 면역형광관찰WAVE1여기질금속단백-2(MMP-2)재세포중적분포.이용PeDNA3.1WAVE1중조진핵표체질립전염K562세포,장WAVE1기인특이성소편단간우RNA(WAVEI siRNA)전염K562세포,Transwell법검측세포침습능력;실시PCR화Western blot검측전염전후WAVE1급MMP-2적표체.결과 ①WAVEI여MMP-2주요표체우K562세포적세포막상차량자유공정위.②여대조조K562세포상비,전염pcDNA3.1-WAVE1 24 h급48 h후K562세포MMP-2 mRNA표체분별증가295%화198%,단백표체수평분별증가80%화23%;전염특이성WAVE1 siRNA 24 h급48 h후K562세포MMP-2 mRNA표체분별하강81%화28%,단백표체분별하조36%화53%.③여대조조K562세포상비전염pcDNA3.1-WAVE1후세포침습능력증강,이간우WAVE1표체후K562세포침습능력감약.결론 WAVE1여MMP-2재K562세포가능구유협동작용;WAVE1삼여료K562세포적침습전이과정,기궤제가능여조공MMP-2적표체상관.
Objective To investigate role of WASP family verprolin homologous protein 1(WAVE1)in K562 leukemia cell invasion and its mechanism.Methods Immunofluoresence methed was used to detect the distribution of WAVE1 and MMP-2 in the cells.K562 cells were transfected with pcDNA3.1-WAVE1 reconstructed plasmid or with specific siRNA to WAVE1 gene.The invasion ability of K562 cells Was examined by Transwell assay.The expression level of WAVE1 and MMP-2 in K562 cells was assayed by real-time PCR by 295%and 198%while its protein increased by 80%and 23%respectively as compared with control K562cells.At the same time point ofter transfected with specific siRNA,the MMP-2 mRNA level decreased by vasion ability of K562 cells was enhanced after transfected with pcDNA3.1-WAVE1 and depressed after transfected with the specific siRNA.Conclusion The co-localization of WAVE1 and MMP-2 in K562 cells suggests they coordinate in functions:WAVE1 may involve in the migration and invasion of K562 cells through regulating the expression level of MMP-2.
