重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2015年
11期
1452-1456
,共5页
肖高春%童仕伦%郑勇斌%郝志楠%李盛波
肖高春%童仕倫%鄭勇斌%郝誌楠%李盛波
초고춘%동사륜%정용빈%학지남%리성파
1-磷脂酰肌醇3-激酶%肿瘤%血管%内皮细胞%细胞运动%信号通路
1-燐脂酰肌醇3-激酶%腫瘤%血管%內皮細胞%細胞運動%信號通路
1-린지선기순3-격매%종류%혈관%내피세포%세포운동%신호통로
1-phosphatidylinositol 3-kinase%neoplasms%blood vessels%endothlial cells%cell movement%signaling pathway
目的:研究磷脂酰肌醇3激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(AKT)及丝裂原细胞外信号调节激酶(MEK)/细胞外信号调节激酶(ERK)信号通路在肿瘤血管内皮细胞迁移中的作用及机制。方法用不同浓度的PI3K/AKT信号通路抑制剂LY294002(2.50、7.50、15.00μmol/L);MEK/ERK信号通路抑制剂PD98059(2.50、7.50、15.00μmol/L)分别处理肿瘤血管内皮细胞,以DMEM‐F12培养基,加入0.10%二甲基亚砜(DMSO)的培养基分别作为对照,通过细胞划痕试验,定向迁移试验和Transwell试验来检测不同的信号通路抑制剂对肿瘤血管内皮细胞的水平、垂直和定向迁移能力的影响。结果0.1%DMSO对内皮细胞的迁移无明显影响,其作用后内皮细胞的迁移能力与DMEM‐F12单独作用无明显区别,说明小剂量DMSO作为LY294002、PD98059的溶剂不影响肿瘤血管内皮细胞的迁移功能;应用信号通路抑制剂LY294002、PD98059处理后,内皮细胞迁移能力受抑制,且随着抑制剂浓度增高而增大;LY294002与PD98059组比较,前者迁移距离更小,细胞迁移数较后者少,二者比较差异有统计学意义(P<0.05)。结论抑制PI3K/AKT和MEK/ERK信号通路均能抑制肿瘤血管内皮细胞的水平、垂直和定向迁移,且与抑制剂浓度呈正相关;PI3K/AKT信号通路对内皮细胞迁移的影响比MEK/ERK信号通路明显。
目的:研究燐脂酰肌醇3激酶(PI3K)/絲氨痠囌氨痠蛋白激酶(AKT)及絲裂原細胞外信號調節激酶(MEK)/細胞外信號調節激酶(ERK)信號通路在腫瘤血管內皮細胞遷移中的作用及機製。方法用不同濃度的PI3K/AKT信號通路抑製劑LY294002(2.50、7.50、15.00μmol/L);MEK/ERK信號通路抑製劑PD98059(2.50、7.50、15.00μmol/L)分彆處理腫瘤血管內皮細胞,以DMEM‐F12培養基,加入0.10%二甲基亞砜(DMSO)的培養基分彆作為對照,通過細胞劃痕試驗,定嚮遷移試驗和Transwell試驗來檢測不同的信號通路抑製劑對腫瘤血管內皮細胞的水平、垂直和定嚮遷移能力的影響。結果0.1%DMSO對內皮細胞的遷移無明顯影響,其作用後內皮細胞的遷移能力與DMEM‐F12單獨作用無明顯區彆,說明小劑量DMSO作為LY294002、PD98059的溶劑不影響腫瘤血管內皮細胞的遷移功能;應用信號通路抑製劑LY294002、PD98059處理後,內皮細胞遷移能力受抑製,且隨著抑製劑濃度增高而增大;LY294002與PD98059組比較,前者遷移距離更小,細胞遷移數較後者少,二者比較差異有統計學意義(P<0.05)。結論抑製PI3K/AKT和MEK/ERK信號通路均能抑製腫瘤血管內皮細胞的水平、垂直和定嚮遷移,且與抑製劑濃度呈正相關;PI3K/AKT信號通路對內皮細胞遷移的影響比MEK/ERK信號通路明顯。
목적:연구린지선기순3격매(PI3K)/사안산소안산단백격매(AKT)급사렬원세포외신호조절격매(MEK)/세포외신호조절격매(ERK)신호통로재종류혈관내피세포천이중적작용급궤제。방법용불동농도적PI3K/AKT신호통로억제제LY294002(2.50、7.50、15.00μmol/L);MEK/ERK신호통로억제제PD98059(2.50、7.50、15.00μmol/L)분별처리종류혈관내피세포,이DMEM‐F12배양기,가입0.10%이갑기아풍(DMSO)적배양기분별작위대조,통과세포화흔시험,정향천이시험화Transwell시험래검측불동적신호통로억제제대종류혈관내피세포적수평、수직화정향천이능력적영향。결과0.1%DMSO대내피세포적천이무명현영향,기작용후내피세포적천이능력여DMEM‐F12단독작용무명현구별,설명소제량DMSO작위LY294002、PD98059적용제불영향종류혈관내피세포적천이공능;응용신호통로억제제LY294002、PD98059처리후,내피세포천이능력수억제,차수착억제제농도증고이증대;LY294002여PD98059조비교,전자천이거리경소,세포천이수교후자소,이자비교차이유통계학의의(P<0.05)。결론억제PI3K/AKT화MEK/ERK신호통로균능억제종류혈관내피세포적수평、수직화정향천이,차여억제제농도정정상관;PI3K/AKT신호통로대내피세포천이적영향비MEK/ERK신호통로명현。
Objective To research the effect and mechanism of phosphatidylinositol 3‐kinase(PI3K)/serine threonine protein kinase(AKT) and mitogen extracellular signal‐regulated kinase(MEK)/extracellular signal‐regulated kinase(ERK) signaling path‐way in tumor vascular endothelial cell migration .Methods The different concentrations of PI3K/AKT signaling pathway inhibitor LY294002 (2 .50 ,7 .50 ,15 .00 μmol/L) and the MEK/ERK signaling pathway inhibitor PD98059 (2 .50 ,7 .50 ,15 .00 μmol/L) were used to treat the tumor‐derived endothelial cells respectively .The DMEM‐F12 culture medium was added into 0 .10% DMSO culture medium as the control .The cell scratch test ,cell directional migration test and Transwell test were adopted to detect the effects of different signaling pathway inhibitors on the tumor vascular endothelial cells level ,horizontal ,vertical and directional mi‐gration .Results 0 .10% DMSO had no significant effect on endothelial cell migration and the role of endothelial cells′migration a‐bility after its action had no obvious difference compared with the single DMEM‐F12 medium action ,indicating that small doses of DMSO as the solvent of LY294002 ,PD98059 did not affect the tumor vascular endothelial cell migration function ;after treatment by signaling pathway inhibitor LY294002 and PD98059 ,the endothelial cell migration ability was suppressed and increased with the in‐hibitor concentration increase ;compared with the PD98059 group ,the migration distance in the LY294002 group was smaller and the number of migrating cells was less ,the differences had statistical significance (P<0 .05) .Conclusion The inhibition of PI3K/AKT and MEK/ERK signaling pathway can inhibit the level of tumor vascular endothelial cells ,vertical and directional migration , and which is positively correlated with the concentration of inhibitor ;the effect of PI3K/AKT signal pathway on the migration of endothelial cells is more significant than that of the MEK/ERK signal pathway .