生物学杂志
生物學雜誌
생물학잡지
JOURNAL OF BIOLOGY
2015年
3期
4-8
,共5页
Kallistatin%HUVEC%NF-κB%侵袭%信号通路
Kallistatin%HUVEC%NF-κB%侵襲%信號通路
Kallistatin%HUVEC%NF-κB%침습%신호통로
kallistatin%HUVEC%NF-κB%invasion%signaling pathway
通过构建NF-κB-p65真核表达质粒,研究NF-κB-p65在重组Kallistatin (rhKal)抑制内皮细胞侵袭中的作用及其分子机制。利用基因重组技术构建NF-κB-p65真核表达质粒载体,转染入HUVEC细胞,检测NF-κB-p65蛋白表达变化;Transwell小室观察重组Kallistatin对转染NF-κB-p65真核表达质粒HUVEC细胞侵袭的影响;Western blotting检测过表达p65对重组Kallistatin对内皮细胞侵袭相关蛋白抑制作用的影响。人脐静脉内皮细胞转染NF-κB-p65真核表达质粒载体后,细胞内p65蛋白表达明显升高;p65过表达可以逆转rhKal对HUVEC细胞体外侵袭的抑制作用;过表达p65可以明显抵抗rhKal对NF-κB信号通路下游相关靶基因的抑制作用。研究表明重组Kallistatin明显抑制HUVEC细胞体外侵袭,其分子机制可能与NF-κB信号通路相关。
通過構建NF-κB-p65真覈錶達質粒,研究NF-κB-p65在重組Kallistatin (rhKal)抑製內皮細胞侵襲中的作用及其分子機製。利用基因重組技術構建NF-κB-p65真覈錶達質粒載體,轉染入HUVEC細胞,檢測NF-κB-p65蛋白錶達變化;Transwell小室觀察重組Kallistatin對轉染NF-κB-p65真覈錶達質粒HUVEC細胞侵襲的影響;Western blotting檢測過錶達p65對重組Kallistatin對內皮細胞侵襲相關蛋白抑製作用的影響。人臍靜脈內皮細胞轉染NF-κB-p65真覈錶達質粒載體後,細胞內p65蛋白錶達明顯升高;p65過錶達可以逆轉rhKal對HUVEC細胞體外侵襲的抑製作用;過錶達p65可以明顯牴抗rhKal對NF-κB信號通路下遊相關靶基因的抑製作用。研究錶明重組Kallistatin明顯抑製HUVEC細胞體外侵襲,其分子機製可能與NF-κB信號通路相關。
통과구건NF-κB-p65진핵표체질립,연구NF-κB-p65재중조Kallistatin (rhKal)억제내피세포침습중적작용급기분자궤제。이용기인중조기술구건NF-κB-p65진핵표체질립재체,전염입HUVEC세포,검측NF-κB-p65단백표체변화;Transwell소실관찰중조Kallistatin대전염NF-κB-p65진핵표체질립HUVEC세포침습적영향;Western blotting검측과표체p65대중조Kallistatin대내피세포침습상관단백억제작용적영향。인제정맥내피세포전염NF-κB-p65진핵표체질립재체후,세포내p65단백표체명현승고;p65과표체가이역전rhKal대HUVEC세포체외침습적억제작용;과표체p65가이명현저항rhKal대NF-κB신호통로하유상관파기인적억제작용。연구표명중조Kallistatin명현억제HUVEC세포체외침습,기분자궤제가능여NF-κB신호통로상관。
The effection and the molecular mechanism of recombinant Kallistatin (rhKal) on the anti-invasion of endothelial cell were studied by transfected NF-κB-p65 eukaryotic expression plasmid. HUVEC cells were transfected by NF-κB-p65 eukaryotic expression vector which was constructed by genetic recombination technology. The expression of p65 protein was also detected by Western blotting. The invasion of transfected HUVEC cells was detected by Transwell chamber after treated with recombinant Kallistatin. The expression of invasion associated protein was detected by Western blotting in endothelial cells which were transfected by NF-κB-p65 expression vector. The results showed that the expression of p65 was significantly increased after transfected with NF-κB-p65 eukaryotic expression vector. Overexpression of p65 gene can reverse the inhibitory of invasion on HUVEC cells by rhKal. Overexpression of p65 gene can significantly increase downstream target genes of NF-κB signaling pathway after treated with rhKal. In conclusion, This study showed that the recombinant Kallistatin could inhibit HUVEC cell invasion in vitro, and its molecular mechanism may be related to the NF-κB signaling pathway.