中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
2期
86-89
,共4页
李伟%龚斐然%陈政%宗洋%陈凯%李道明%陶敏%徐泽宽
李偉%龔斐然%陳政%宗洋%陳凱%李道明%陶敏%徐澤寬
리위%공비연%진정%종양%진개%리도명%도민%서택관
蛋白质磷酸酶2%胰腺肿瘤%JNK信号通路%c-Jun%AP-1
蛋白質燐痠酶2%胰腺腫瘤%JNK信號通路%c-Jun%AP-1
단백질린산매2%이선종류%JNK신호통로%c-Jun%AP-1
Protein phosphatase 2%Pancreatic neoplasms%JNK%c-Jun%AP-1
目的 探讨蛋白磷酸酶2A催化性C亚基(PP2 Ac)在胰腺癌组织和细胞系中的表达水平,及其对JNK/c-Jun/AP-1信号通路的调控作用和对癌细胞生长的影响.方法 采用实时PCR检测PP2Ac表达水平,MTT法检测细胞活力,Western印迹检测蛋白表达水平和磷酸化水平,流式细胞术检测细胞周期,荧光素酶报告基因检测转录活性.结果 胰腺癌组织中PP2Ac表达水平显著低于癌旁组织,胰腺癌细胞系中PP2Ac也呈低水平表达,在胰腺癌细胞中过表达PP2Ac可抑制癌细胞生长(转染PP2Acα、PP2Acβ 48 h后可使细胞活力分别下降(24.85±4.85)％、(11.31±4.62)％;72 h后细胞活力分别下降(33.89±2.05)％、(16.66±2.81)％;PP2Ac低水平表达可上调JNK/c-Jun/AP-1通路的活性,在胰腺癌细胞中过表达PP2Ac可下调JNK、c-Jun的磷酸化水平并抑制AP-1的转录活性;采用抑制剂阻断JNK通路,可阻滞细胞周期于G2/M期,进而抑制细胞生长.结论 胰腺癌细胞中PP2Ac呈低表达,通过上调JNK/c-Jun/AP-1通路的活性促进胰腺癌细胞生长.
目的 探討蛋白燐痠酶2A催化性C亞基(PP2 Ac)在胰腺癌組織和細胞繫中的錶達水平,及其對JNK/c-Jun/AP-1信號通路的調控作用和對癌細胞生長的影響.方法 採用實時PCR檢測PP2Ac錶達水平,MTT法檢測細胞活力,Western印跡檢測蛋白錶達水平和燐痠化水平,流式細胞術檢測細胞週期,熒光素酶報告基因檢測轉錄活性.結果 胰腺癌組織中PP2Ac錶達水平顯著低于癌徬組織,胰腺癌細胞繫中PP2Ac也呈低水平錶達,在胰腺癌細胞中過錶達PP2Ac可抑製癌細胞生長(轉染PP2Acα、PP2Acβ 48 h後可使細胞活力分彆下降(24.85±4.85)％、(11.31±4.62)％;72 h後細胞活力分彆下降(33.89±2.05)％、(16.66±2.81)％;PP2Ac低水平錶達可上調JNK/c-Jun/AP-1通路的活性,在胰腺癌細胞中過錶達PP2Ac可下調JNK、c-Jun的燐痠化水平併抑製AP-1的轉錄活性;採用抑製劑阻斷JNK通路,可阻滯細胞週期于G2/M期,進而抑製細胞生長.結論 胰腺癌細胞中PP2Ac呈低錶達,通過上調JNK/c-Jun/AP-1通路的活性促進胰腺癌細胞生長.
목적 탐토단백린산매2A최화성C아기(PP2 Ac)재이선암조직화세포계중적표체수평,급기대JNK/c-Jun/AP-1신호통로적조공작용화대암세포생장적영향.방법 채용실시PCR검측PP2Ac표체수평,MTT법검측세포활력,Western인적검측단백표체수평화린산화수평,류식세포술검측세포주기,형광소매보고기인검측전록활성.결과 이선암조직중PP2Ac표체수평현저저우암방조직,이선암세포계중PP2Ac야정저수평표체,재이선암세포중과표체PP2Ac가억제암세포생장(전염PP2Acα、PP2Acβ 48 h후가사세포활력분별하강(24.85±4.85)％、(11.31±4.62)％;72 h후세포활력분별하강(33.89±2.05)％、(16.66±2.81)％;PP2Ac저수평표체가상조JNK/c-Jun/AP-1통로적활성,재이선암세포중과표체PP2Ac가하조JNK、c-Jun적린산화수평병억제AP-1적전록활성;채용억제제조단JNK통로,가조체세포주기우G2/M기,진이억제세포생장.결론 이선암세포중PP2Ac정저표체,통과상조JNK/c-Jun/AP-1통로적활성촉진이선암세포생장.
Objective To investigate the expression of protein phosphatase 2A catalytic subunit (PP2Ac) in pancreatic cancer and the regulation of this gene on JNK/c-Jun/AP-1 pathway and cell growth.Methods Expression of PP2Ac was determined by real-time PCR.Cell viability was tested by MTT.Expression and phosphorylation levels of proteins were detected by Western blotting.Cell cycle was assayed by flow cytometry.Transcription activity was measured by luciferase reporter gene assay.Results Suppressed PP2Ac expression was detected in pancreatic cancer tissues.PP2Ac expression in the pancreatic cancer cell lines was also at a low level.Overexpression of the two isoforms of PP2Ac,PP2Acα and PP2Acβ,in pancreatic cancer cells repressed cell growth.Cell viability decreased (33.89 ± 2.05) ％ (t =28.607,P＜0.001) and (16.66 ± 2.81)％ (t =10.257,P =0.001) respectively 72 hours after transfection.Overexpression of PP2Acα and PP2Acβ down-regulated the phosphorylation levels of JNK and cJun,and made the transcriptional activity of AP-1 decrease (47.18 ±2.28) ％ (t =11.230,P ＜0.001) and (30.89 ± 8.09) ％ (t =6.612,P =0.003) respectively,indicating the down-regulation of PP2Ac upregulated the activity of JNK/c-Jun/AP-1 pathway.Blocking the JNK pathway using a selective inhibitor,SP600125,induced G2/M cell cycle arrest and repressed cell proliferation.Cell viability decreased (31.38 ±1.33) ％ (t =40.930,P ＜ 0.001) after treatment with JNK inhibitor for 72 hours.Conclusion Suppressed expression of PP2Ac in pancreatic cancer facilitated cell growth through up-regulating the activity of JNK/cJun/AP-1 pathway.
