中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2013年
3期
229-233
,共5页
刘勇%谭浩蕾%李果%余长云%粟忠武%任舒灵%朱刚才%邱元正%田勇泉
劉勇%譚浩蕾%李果%餘長雲%粟忠武%任舒靈%硃剛纔%邱元正%田勇泉
류용%담호뢰%리과%여장운%속충무%임서령%주강재%구원정%전용천
受体,EphA2%头颈部肿瘤%癌,鳞状细胞%p38丝裂原活化蛋白激酶类%新生血管化,病理性%血管内皮生长因子A
受體,EphA2%頭頸部腫瘤%癌,鱗狀細胞%p38絲裂原活化蛋白激酶類%新生血管化,病理性%血管內皮生長因子A
수체,EphA2%두경부종류%암,린상세포%p38사렬원활화단백격매류%신생혈관화,병이성%혈관내피생장인자A
Receptor,EphA2%Head and neck neoplasms%Carcinoma,Squamous cell%p38mitogen-activated protein kinases%Neovascularization,pathologic%Vascular endothelial growth factor A
目的 阐明促红细胞生成素产生肝细胞受体A2(erythropoietin-producing hepatocellular receptor,EphA2)蛋白可经p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)信号通路调控头颈部鳞状细胞癌血管内皮细胞生长因子(VEGF)的表达.方法 采用脂质体2000将EphA2蛋白过表达载体pEGFP-N1-EphA2和空白载体pEGFP-N1分别转染头颈鳞状细胞癌Tu686细胞,酶联免疫吸附法检测EphA2蛋白上调后VEGF蛋白的表达,Western blot法检测p38 MAPK信号通路有无激活；进一步利用不同浓度(0、5、10 μmol/L)小分子抑制剂SB203580特异性阻断p38MAPK通路,酶联免疫吸附法检测VEGF蛋白的表达改变有无逆转.结果 Tu686细胞EphA2蛋白上调后,VEGF蛋白在空白载体组和亲本细胞组中的表达(x±s)分别为(400.99±33.50) pg/ml和(385.30±33.50) pg/ml,而在实验组中的表达显著升高为(535.31±45.71) pg/ml,差异有统计学意义(F=17.091,P＜0.01)；同时,实验组Tu686细胞磷酸化p38 MAPK的表达上调.不同浓度(0、5、10 μmol/L) SB203580作用EphA2蛋白过表达的Tu686细胞后,磷酸化p38 MAPK的表达则被梯度抑制,且VEGF蛋白的表达逐渐降低,分别为(643.75±52.00) pg/ml、(513.52±54.05) pg/ml、(302.85±13.93) pg/ml,三组间差异有统计学意义(F=45.761,P＜0.01).结论 EphA2蛋白对p38MAPK信号通路具有调控作用,并经该通路影响VEGF蛋白的表达.
目的 闡明促紅細胞生成素產生肝細胞受體A2(erythropoietin-producing hepatocellular receptor,EphA2)蛋白可經p38絲裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)信號通路調控頭頸部鱗狀細胞癌血管內皮細胞生長因子(VEGF)的錶達.方法 採用脂質體2000將EphA2蛋白過錶達載體pEGFP-N1-EphA2和空白載體pEGFP-N1分彆轉染頭頸鱗狀細胞癌Tu686細胞,酶聯免疫吸附法檢測EphA2蛋白上調後VEGF蛋白的錶達,Western blot法檢測p38 MAPK信號通路有無激活；進一步利用不同濃度(0、5、10 μmol/L)小分子抑製劑SB203580特異性阻斷p38MAPK通路,酶聯免疫吸附法檢測VEGF蛋白的錶達改變有無逆轉.結果 Tu686細胞EphA2蛋白上調後,VEGF蛋白在空白載體組和親本細胞組中的錶達(x±s)分彆為(400.99±33.50) pg/ml和(385.30±33.50) pg/ml,而在實驗組中的錶達顯著升高為(535.31±45.71) pg/ml,差異有統計學意義(F=17.091,P＜0.01)；同時,實驗組Tu686細胞燐痠化p38 MAPK的錶達上調.不同濃度(0、5、10 μmol/L) SB203580作用EphA2蛋白過錶達的Tu686細胞後,燐痠化p38 MAPK的錶達則被梯度抑製,且VEGF蛋白的錶達逐漸降低,分彆為(643.75±52.00) pg/ml、(513.52±54.05) pg/ml、(302.85±13.93) pg/ml,三組間差異有統計學意義(F=45.761,P＜0.01).結論 EphA2蛋白對p38MAPK信號通路具有調控作用,併經該通路影響VEGF蛋白的錶達.
목적 천명촉홍세포생성소산생간세포수체A2(erythropoietin-producing hepatocellular receptor,EphA2)단백가경p38사렬원활화단백격매(p38 mitogen-activated protein kinase,p38 MAPK)신호통로조공두경부린상세포암혈관내피세포생장인자(VEGF)적표체.방법 채용지질체2000장EphA2단백과표체재체pEGFP-N1-EphA2화공백재체pEGFP-N1분별전염두경린상세포암Tu686세포,매련면역흡부법검측EphA2단백상조후VEGF단백적표체,Western blot법검측p38 MAPK신호통로유무격활；진일보이용불동농도(0、5、10 μmol/L)소분자억제제SB203580특이성조단p38MAPK통로,매련면역흡부법검측VEGF단백적표체개변유무역전.결과 Tu686세포EphA2단백상조후,VEGF단백재공백재체조화친본세포조중적표체(x±s)분별위(400.99±33.50) pg/ml화(385.30±33.50) pg/ml,이재실험조중적표체현저승고위(535.31±45.71) pg/ml,차이유통계학의의(F=17.091,P＜0.01)；동시,실험조Tu686세포린산화p38 MAPK적표체상조.불동농도(0、5、10 μmol/L) SB203580작용EphA2단백과표체적Tu686세포후,린산화p38 MAPK적표체칙피제도억제,차VEGF단백적표체축점강저,분별위(643.75±52.00) pg/ml、(513.52±54.05) pg/ml、(302.85±13.93) pg/ml,삼조간차이유통계학의의(F=45.761,P＜0.01).결론 EphA2단백대p38MAPK신호통로구유조공작용,병경해통로영향VEGF단백적표체.
Objective To investigate the regulatory effect of erythropoietin-producing hepatocellular receptor (EphA2) on the expression of VEGF protein,a pro-angiogenic factor,via p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in squamous cell carcinoma of the head and neck(SCCHN)in vitro.Methods SCCHN Tu686 cells were transfected with EphA2 overexpression vector pEGFP-N1-EphA2.Western blot was used to detect the expression of p38 MAPK and enzyme-linked immunosorbent assay(ELISA) was applied to assay of VEGF.SB203580 as a inhibitor of p38 MAPK signaling pathway was used.Results The expression of VEGF protein was significantly up-regulated in Tu686 cells transfected with EphA2 overexpression vector (535.31 ±45.71) pg/ml,when compared with Tu686 cells transfected with empty vector (400.99 ± 33.50) pg/ml and Tu686 cells with no transfection (385.30 ± 33.50) pg/ml (F=17.091,P ＜0.01).The expression of phosphorylated p38 MAPK was obviously increased in Tu686 cells with EphA2 overexpression.SB203580 inhibited the expressions of VEGF and phosphorylated p38 MAPK proteins in Tu686 cells with EphA2 overexpression.Conclusion EphA2 can regulate the expression of VEGF protein and stimulate p38 MAPK signaling pathway.
