现代医药卫生
現代醫藥衛生
현대의약위생
MODERN MEDICINE HEALTH
2014年
7期
963-965
,共3页
孙晓东%王珺%王燕%田宗文
孫曉東%王珺%王燕%田宗文
손효동%왕군%왕연%전종문
血管内皮生长因子A%酶联免疫吸附测定%RNA干扰%DLL4/Notch信号途径%A549细胞
血管內皮生長因子A%酶聯免疫吸附測定%RNA榦擾%DLL4/Notch信號途徑%A549細胞
혈관내피생장인자A%매련면역흡부측정%RNA간우%DLL4/Notch신호도경%A549세포
Vascular endothelial growth factor A%Enzyme-linked immunosorbent assay%RNA interference%DLL4/notch signal pathway%A549 cells
目的：研究Delta-like ligand 4（DLL4）与血管内皮生长因子（VEGF）在人肺腺癌A549细胞中的表达，以及DLL4对VEGF表达的影响。方法（1）用免疫组化检测DLL4与VEGF在A549细胞中的表达；（2）设计和化学合成靶向DLL4基因的小干扰RNA（siRNA）序列，用lipofectamineTM2000介导DLL4 siRNA转染人A549细胞；（3）提取细胞总RNA，进行反转录-聚合酶链反应（RT-PCR）扩增检测DLL4、VEGF mRNA；（4）用酶联免疫吸附试验（ELISA）检测A549细胞培养上清液中VEGF质量浓度。结果（1）DLL4、VEGF表达于A549细胞的细胞质内；（2）DLL4 siRNA可使DLL4沉默，同时，明显上调VEGF-A、VEGF-B、VEGF-C mRNA表达，差异有统计学意义（P<0.05）；（3）DLL4转染后，细胞培养上清液中VEGF质量浓度明显增加，差异有统计学意义（P<0.01）。结论阻断A549细胞中Notch通路可刺激VEGF过表达。
目的：研究Delta-like ligand 4（DLL4）與血管內皮生長因子（VEGF）在人肺腺癌A549細胞中的錶達，以及DLL4對VEGF錶達的影響。方法（1）用免疫組化檢測DLL4與VEGF在A549細胞中的錶達；（2）設計和化學閤成靶嚮DLL4基因的小榦擾RNA（siRNA）序列，用lipofectamineTM2000介導DLL4 siRNA轉染人A549細胞；（3）提取細胞總RNA，進行反轉錄-聚閤酶鏈反應（RT-PCR）擴增檢測DLL4、VEGF mRNA；（4）用酶聯免疫吸附試驗（ELISA）檢測A549細胞培養上清液中VEGF質量濃度。結果（1）DLL4、VEGF錶達于A549細胞的細胞質內；（2）DLL4 siRNA可使DLL4沉默，同時，明顯上調VEGF-A、VEGF-B、VEGF-C mRNA錶達，差異有統計學意義（P<0.05）；（3）DLL4轉染後，細胞培養上清液中VEGF質量濃度明顯增加，差異有統計學意義（P<0.01）。結論阻斷A549細胞中Notch通路可刺激VEGF過錶達。
목적：연구Delta-like ligand 4（DLL4）여혈관내피생장인자（VEGF）재인폐선암A549세포중적표체，이급DLL4대VEGF표체적영향。방법（1）용면역조화검측DLL4여VEGF재A549세포중적표체；（2）설계화화학합성파향DLL4기인적소간우RNA（siRNA）서렬，용lipofectamineTM2000개도DLL4 siRNA전염인A549세포；（3）제취세포총RNA，진행반전록-취합매련반응（RT-PCR）확증검측DLL4、VEGF mRNA；（4）용매련면역흡부시험（ELISA）검측A549세포배양상청액중VEGF질량농도。결과（1）DLL4、VEGF표체우A549세포적세포질내；（2）DLL4 siRNA가사DLL4침묵，동시，명현상조VEGF-A、VEGF-B、VEGF-C mRNA표체，차이유통계학의의（P<0.05）；（3）DLL4전염후，세포배양상청액중VEGF질량농도명현증가，차이유통계학의의（P<0.01）。결론조단A549세포중Notch통로가자격VEGF과표체。
Objective To study the expression of Delta-like ligand 4 ( DLL4 ) and vascular endothelial growth factor (VEGF) in A549 cells of human lung adenocarcinoma,and the influence of DLL4 on the expression of VEGF. Methods (1) The immunohistochemical method was adopted to detect the expression of DLL4 and VEGF in A549 cells;(2)the small interferon RNA(siRNA) squences of targeted DLL4 gene were designed and chemically synthesized,and the DLL4 siRNAs were transfect-ed into A549 cells by lipofectamineTM2000;(3)total RNA was extracted from A549 cells to conduct reversal transcription poly merase chain reaction(RT-PCR) for amplifying and detecting DLL4 and VEGF mRNA;(4)the enzyme linked immunosorbent assay(ELISA) were applied to determine the mass concentration of VEGF in culture supernatant fluid of A549 cells. Results (1)DLL4 and VEGF were expressed in cytoplasm of A549 cells;(2)the siRAN of DLL4 gene could make DLL4 gene silence,and up-regulate the expression of VEGF-A mRNA,VEGF-B mRNA amd VEGF-C mRNA with statistically significant difference (P<0.05);(3)after transfection of DLL4 cells,the mass concentration of VEGF in culture supernatant fluid increased obviously with statistically sig-nificant difference(P<0.01). Conclusion Blocking Notch signal pathway in A549 cells can stimulate the expression of VEGF.
