中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1911-1914
,共4页
肖高春%郑勇斌%童仕伦%郝志楠%李盛波
肖高春%鄭勇斌%童仕倫%郝誌楠%李盛波
초고춘%정용빈%동사륜%학지남%리성파
结肠癌相关血管内皮细胞%增殖%迁移%侵袭
結腸癌相關血管內皮細胞%增殖%遷移%侵襲
결장암상관혈관내피세포%증식%천이%침습
Colon cancer-related vascular endothelial cells%Proliferation%Migartion%Invasion
目的 观察磷脂酰肌醇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)信号通路在结肠癌血管形成过程中对结肠癌相关血管内皮细胞(CCRVEC)功能的影响.方法 用不同浓度(2.5、7.5、15.0 μmol/L)的信号通路抑制剂分别干预CCRVEC的PI3K/Akt和MEK/ERK信号通路,通过细胞计数法、细胞划痕、定向迁移、Transwell迁移、侵袭和管道形成实验检测不同的信号通路对CCRVEC的增殖、迁移、侵袭与管道形成能力的影响.结果 抑制CCRVEC的PI3 K/Akt和MEK/ERK信号通路后,其增殖、迁移、侵袭及管道形成能力均受抑制,且该效应随着抑制剂浓度增高而呈增强趋势;15 μmol/L的PI3 K/Akt和MEK/ERK信号通路抑制剂分别处理CCRVEC 6 h后,其管道形成长度为(1.28±0.13) mm和(1.87 ±0.13)mm,明显短于对照组[(4.07±0.26) mm,P＜0.05];24h后,侵袭细胞数为(207.38±24.29)、(264.02±15.54)个,少于对照组[(475.81±16.66)个,P＜0.05];细胞迁移数分别为(229.50±20.82)、(271.12±16.10)、(472.33±11.83)个(P ＜0.05);30 h后,各组间细胞迁移距离差异有统计学意义(P＜0.05);抑制剂处理6d后细胞增殖数分别为(56.29±3.85)、(46.23±3.31)和(91.23±4.82)个/孔(P＜0.05);相同的抑制剂浓度抑制PI3K/Akt对细胞迁移、侵袭和管道形成抑制作用较MEK/ERK通路抑制效应更明显,但MEK/ERK信号通路对CCRVEC增殖的影响较PI3 K/Akt信号通路明显(P＜0.05).结论 PI3K/Akt和MEK/ERK信号通路均参与结肠癌血管形成中的增殖、迁移、侵袭及管道形成过程;与PI3K/Akt信号通路主要影响CCRVEC迁移、侵袭与管道形成等过程不同,MEK/ERK信号通路尽管也影响CCRVEC迁移、侵袭及管道形成,但对增殖的影响则更明显.
目的 觀察燐脂酰肌醇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)信號通路在結腸癌血管形成過程中對結腸癌相關血管內皮細胞(CCRVEC)功能的影響.方法 用不同濃度(2.5、7.5、15.0 μmol/L)的信號通路抑製劑分彆榦預CCRVEC的PI3K/Akt和MEK/ERK信號通路,通過細胞計數法、細胞劃痕、定嚮遷移、Transwell遷移、侵襲和管道形成實驗檢測不同的信號通路對CCRVEC的增殖、遷移、侵襲與管道形成能力的影響.結果 抑製CCRVEC的PI3 K/Akt和MEK/ERK信號通路後,其增殖、遷移、侵襲及管道形成能力均受抑製,且該效應隨著抑製劑濃度增高而呈增彊趨勢;15 μmol/L的PI3 K/Akt和MEK/ERK信號通路抑製劑分彆處理CCRVEC 6 h後,其管道形成長度為(1.28±0.13) mm和(1.87 ±0.13)mm,明顯短于對照組[(4.07±0.26) mm,P＜0.05];24h後,侵襲細胞數為(207.38±24.29)、(264.02±15.54)箇,少于對照組[(475.81±16.66)箇,P＜0.05];細胞遷移數分彆為(229.50±20.82)、(271.12±16.10)、(472.33±11.83)箇(P ＜0.05);30 h後,各組間細胞遷移距離差異有統計學意義(P＜0.05);抑製劑處理6d後細胞增殖數分彆為(56.29±3.85)、(46.23±3.31)和(91.23±4.82)箇/孔(P＜0.05);相同的抑製劑濃度抑製PI3K/Akt對細胞遷移、侵襲和管道形成抑製作用較MEK/ERK通路抑製效應更明顯,但MEK/ERK信號通路對CCRVEC增殖的影響較PI3 K/Akt信號通路明顯(P＜0.05).結論 PI3K/Akt和MEK/ERK信號通路均參與結腸癌血管形成中的增殖、遷移、侵襲及管道形成過程;與PI3K/Akt信號通路主要影響CCRVEC遷移、侵襲與管道形成等過程不同,MEK/ERK信號通路儘管也影響CCRVEC遷移、侵襲及管道形成,但對增殖的影響則更明顯.
목적 관찰린지선기순3격매(PI3K)/사안산소안산단백격매(Akt)급사렬원세포외신호조절격매(MEK)/세포외신호조절격매(ERK)신호통로재결장암혈관형성과정중대결장암상관혈관내피세포(CCRVEC)공능적영향.방법 용불동농도(2.5、7.5、15.0 μmol/L)적신호통로억제제분별간예CCRVEC적PI3K/Akt화MEK/ERK신호통로,통과세포계수법、세포화흔、정향천이、Transwell천이、침습화관도형성실험검측불동적신호통로대CCRVEC적증식、천이、침습여관도형성능력적영향.결과 억제CCRVEC적PI3 K/Akt화MEK/ERK신호통로후,기증식、천이、침습급관도형성능력균수억제,차해효응수착억제제농도증고이정증강추세;15 μmol/L적PI3 K/Akt화MEK/ERK신호통로억제제분별처리CCRVEC 6 h후,기관도형성장도위(1.28±0.13) mm화(1.87 ±0.13)mm,명현단우대조조[(4.07±0.26) mm,P＜0.05];24h후,침습세포수위(207.38±24.29)、(264.02±15.54)개,소우대조조[(475.81±16.66)개,P＜0.05];세포천이수분별위(229.50±20.82)、(271.12±16.10)、(472.33±11.83)개(P ＜0.05);30 h후,각조간세포천이거리차이유통계학의의(P＜0.05);억제제처리6d후세포증식수분별위(56.29±3.85)、(46.23±3.31)화(91.23±4.82)개/공(P＜0.05);상동적억제제농도억제PI3K/Akt대세포천이、침습화관도형성억제작용교MEK/ERK통로억제효응경명현,단MEK/ERK신호통로대CCRVEC증식적영향교PI3 K/Akt신호통로명현(P＜0.05).결론 PI3K/Akt화MEK/ERK신호통로균삼여결장암혈관형성중적증식、천이、침습급관도형성과정;여PI3K/Akt신호통로주요영향CCRVEC천이、침습여관도형성등과정불동,MEK/ERK신호통로진관야영향CCRVEC천이、침습급관도형성,단대증식적영향칙경명현.
Objective To observe the effect of phosphatidylinositol 3-kinase (PI3K)/serine/ threonine protein kinase (Akt) and mitogen extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway on colon cancer-related vascular endothelial cells (CCRVECs) function in the process of the tumor angiogenesis in colon cancer.Methods CCRVECs were treated with different concentrations (2.5,7.5,15.0 μmol/L) inhibitors of PI3K/Akt and MEK/ERK signal pathway.The proliferation of CCRVECs was measured by cell counting.The cell migration and invasion ability was tested by cell scratch,orientation migration,Transwell assays,respectively.simultaneously,tube formation technique was also performed to detect the tube formation on CCRVECs.Results In CCRVECs,following blocking PI3K/Akt and MEK/ERK signal pathway with special inhibitor,the ability of proliferation,migration and invasion was inhibited.Moreover,the inhibiting effect was increased with the increase of the concentration.After CCVECs were exposed to 15 μmol/L of PI3K/Akt and MEK/ERK inhibitors for 6 h,the length of tube formation in three groups was (1.28 ± 0.13) mm,(1.87 ± 0.13) mm and (4.07 ± 0.26) mm respectively,and the length in experiment groups was decreased notably as compared with control group (P ＜ 0.05).At 24 h after treatment with special inhibitor,the number of invasive cells in control group was 475.81 ± 16.66,and that in treated groups was 207.38 ± 24.29 and 264.02 ± 15.54 respectively,with the difference being significant between experiment groups and the control group (P ＜ 0.05).After treatment for 30 h,the cell migration distance showed significant difference among the groups (P ＜ 0.05).After CCVECs were exposed to inhibitor for 6 days,the number of proliferation cells in each well in treated groups and control group was 56.29 ±3.85,46.23 ±3.31 and 91.23 ± 4.82 respectively with the difference being significant among the groups (P ＜ 0.05).The inhibition effect of PI3 K/Akt signal inhibitor was superior to that of MEK/ERK inhibitor in cell migration,invasion,tube formation ability at the same concentration,except cell proliferation ability.Conclusion PI3K/Akt and MEK/ERK signaling pathway participate in the process of the cell proliferation,migration,invasion and tube formation of CCVECs in colon cancer.Unlike PI3K/Akt signaling pathway having main effects on migration,invasion and tune formation of CCVECs,MEK/ERK signaling pathway has more significant influence on cell proliferation of CCVECs.
