CAJ | 학술논문

目的观察受体酪氨酸激酶(Axl)在肝癌细胞转移过程中的作用及其对磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)-p21激活激酶-1(PAK1)信号通路的调控作用.方法体外培养肝癌细胞株MHCC97-H和MHCC97-L细胞.运用Western blot方法研究Axl在肝癌细胞MHCC97-H和MHCC97-L细胞株中的蛋白表达水平.通过短发夹RNA(shRNA)转染肝癌细胞MHCC97-H细胞株抑制Axl的表达,探究Axl对PI3 K/Akt-PAK1信号通路的调控作用机制,并分析Axl蛋白表达与肝癌细胞侵袭与转移的关系.结果与MHCC97-L细胞株比较,Axl在MHCC97-H细胞株中具有更高的表达水平.MHCC97-H细胞中Axl表达下调降低了PI3K、Akt、PAK1信号通路分子的磷酸化水平,并明显抑制了肝癌细胞的侵袭性;侵袭实验结果示:Axl-短发卡RNA(shRNA)转染组穿过基质胶的细胞数为(23±3)个,较质控组的(37±5)个和正常细胞MHCC97-H组的(38±5)个明显减少(P＜0.05).用PI3K特异性抑制剂LY294002或Akt-小干扰RNA (siRNA)阻断PI3K/Akt信号通路明显抑制PAK1的激活及细胞的侵袭性;侵袭实验结果显示:Akt-siRNA转染组和PI3K特异性性抑制剂LY294002抑制组穿过基质胶的细胞数为(23±3)、(18±3)个,较质控组的(37±5)个,二甲基亚砜(DMSO)组的(38±5)个明显减少(P＜0.05).结论 Axl能够通过PI3 K/Akt-PAK1调节肝癌细胞的转移过程.
목적 관찰수체락안산격매(Axl)재간암세포전이과정중적작용급기대린산기순3격매(PI3K)/단백격매B(Akt)-p21격활격매-1(PAK1)신호통로적조공작용.방법 체외배양간암세포주MHCC97-H화MHCC97-L세포.운용Western blot방법연구Axl재간암세포MHCC97-H화MHCC97-L세포주중적단백표체수평.통과단발협RNA(shRNA)전염간암세포MHCC97-H세포주억제Axl적표체,탐구Axl대PI3 K/Akt-PAK1신호통로적조공작용궤제,병분석Axl단백표체여간암세포침습여전이적관계.결과 여MHCC97-L세포주비교,Axl재MHCC97-H세포주중구유경고적표체수평.MHCC97-H세포중Axl표체하조강저료PI3K、Akt、PAK1신호통로분자적린산화수평,병명현억제료간암세포적침습성;침습실험결과시:Axl-단발잡RNA(shRNA)전염조천과기질효적세포수위(23±3)개,교질공조적(37±5)개화정상세포MHCC97-H조적(38±5)개명현감소(P＜0.05).용PI3K특이성억제제LY294002혹Akt-소간우RNA (siRNA)조단PI3K/Akt신호통로명현억제PAK1적격활급세포적침습성;침습실험결과현시:Akt-siRNA전염조화PI3K특이성성억제제LY294002억제조천과기질효적세포수위(23±3)、(18±3)개,교질공조적(37±5)개,이갑기아풍(DMSO)조적(38±5)개명현감소(P＜0.05).결론 Axl능구통과PI3 K/Akt-PAK1조절간암세포적전이과정.
Objective To investigate the possible role of receptotyrosine kinase (Axl) in the metastasiprocesof hepatocellulacarcinomcelland clarify itmodulating mechanism in regulating phosphatidylinositol-3-kinase (PI3K)/protein kinase (Akt)-p21-activated kinases-1 (PAK1) signal pathway.MethodThe hepatocellulacarcinomcell lineMHCC97-H and MHCC97-L cellwere cultured.To evaluate the protein expression levelof Axl in MHCC97-H and MHCC97-L celllineby western bloanalysiand to observe theidifference.Axl shorhairpin RN(shRNA) watransfected into cellto knock down Axl, and then we studied the modulating mechanism of Axl regulating PI3K/Akt-PAK1 signaling pathway.The relationship between the expression role of Axl protein and the invasivenesof hepatocellulacarcinomcellwaclarified.ResultThe resultshowed thathe expression of Axl wahighein MHCC97-H cell linecompared with MHCC97-L cell lines.The invasion ability of MHCC97-H cellwareduced by down regulating the expression of Axl in MHCC97-H cell lines.The experimental resultshowd thatransfected with Axl-shRNafteeach perspective transfecell(23 ± 3), compared with the control group (37 ± 5) and MHCC97-H group (38 ± 5) had significantly decreased (P ＜ 0.05).We also observed thathe PAK1 activation and cell invasion were inhibited by blocking PI3K/Aksignaling pathway by LY294002 oAkt-small interfering RN(siRNA).Invasion and experimental resultshowd that, the celltreated with Akt-siRNand LY294002 through the Matrigel cell numbe(23 ±3, 18 ±3), compared with the control group (37 ±5) and DMSO group (38 ±5), which significantly decreased (P ＜ 0.05).Conclusion These resultprovide insightinto the function of Axl.Axl acting atumometastasis-associated gene can regulate the metastasiprocesof hepatocellulacarcinomcellviPI3K/Akt-PAK1 signal pathway.