CAJ | 학술논문

目的观察过氧化物酶体增殖活化受体β/δ(PPARβ/δ)表达在非小细胞肺癌中的作用.方法逆转录聚合酶链反应(RT-PCR)、荧光素酶报告基因检测非小细胞肺癌(NSCLC)细胞株及肿瘤组织、正常组组织PPARβ/δ、PPARγ、磷脂酶A2(PLA2)、环氧合酶-2(Cox-2)、前列腺素合酶(PGES)、前列环素合酶(PGIS)的表达水平,基因敲除技术及流式细胞仪分析PPARβ/γ对非小细胞癌细胞增殖与存活的影响,药物综合分析PPARβ/γ对Cox-2与血管内皮生长因子(VEGF)表达的影响.结果 H441细胞PPARβ/δ、PPARc、cPLA2、Cox-2、PGES及PPARγ表达水平很高,H358、H23细胞,PPARβ/δ表达水平居中,肿瘤组织PPARβ/δ mRNA水平比正常肺组织要高,PPARβ/δ在A549细胞中活性仅为2.3,而在H441细胞中高达6.1,两者差异有统计学意义(P＜0.01),PPARβ/δ具有促进NSCLC细胞增殖与存活的功能,cPGI2(低浓度,能激活PPARβ/δ)能促进细胞增殖、增加细胞稳定性、延长细胞S期、缩短G1期,但对A549细胞几乎没有影响,当cPGI2高达10 mg/L时,对H441细胞及H358细胞稳定性有很大的促进作用,稳定性分别高达145和151;siRNA敲除PPARβ/δ影响细胞稳定性,促进细胞凋亡,细胞活力与对照组比较,下降到90％;PPARβ/δ影响Cox-2及VEGF的表达,GW501516(PPARβ/δ的配体)处理NSCLC细胞,Cox-2及VEGF mRNA水平有所上升,但是A549细胞变化不明显.结论 PPARβ/δ在NSCLC细胞株及肿瘤组织表达量较高,具有促进NSCLC细胞增殖和稳定性、调节NSCLC细胞周期等功能,并影响Cox-2/VEGF的表达.
목적 관찰과양화물매체증식활화수체β/δ(PPARβ/δ)표체재비소세포폐암중적작용.방법 역전록취합매련반응(RT-PCR)、형광소매보고기인검측비소세포폐암(NSCLC)세포주급종류조직、정상조조직PPARβ/δ、PPARγ、린지매A2(PLA2)、배양합매-2(Cox-2)、전렬선소합매(PGES)、전렬배소합매(PGIS)적표체수평,기인고제기술급류식세포의분석PPARβ/γ대비소세포암세포증식여존활적영향,약물종합분석PPARβ/γ대Cox-2여혈관내피생장인자(VEGF)표체적영향.결과 H441세포PPARβ/δ、PPARc、cPLA2、Cox-2、PGES급PPARγ표체수평흔고,H358、H23세포,PPARβ/δ표체수평거중,종류조직PPARβ/δ mRNA수평비정상폐조직요고,PPARβ/δ재A549세포중활성부위2.3,이재H441세포중고체6.1,량자차이유통계학의의(P＜0.01),PPARβ/δ구유촉진NSCLC세포증식여존활적공능,cPGI2(저농도,능격활PPARβ/δ)능촉진세포증식、증가세포은정성、연장세포S기、축단G1기,단대A549세포궤호몰유영향,당cPGI2고체10 mg/L시,대H441세포급H358세포은정성유흔대적촉진작용,은정성분별고체145화151;siRNA고제PPARβ/δ영향세포은정성,촉진세포조망,세포활력여대조조비교,하강도90％;PPARβ/δ영향Cox-2급VEGF적표체,GW501516(PPARβ/δ적배체)처리NSCLC세포,Cox-2급VEGF mRNA수평유소상승,단시A549세포변화불명현.결론 PPARβ/δ재NSCLC세포주급종류조직표체량교고,구유촉진NSCLC세포증식화은정성、조절NSCLC세포주기등공능,병영향Cox-2/VEGF적표체.
Objective To investigate the function of peroxisome proliferator-activated receptor β/δ (PPARβ/δ) expression in non-small cell lung cancer (NSCLC).Methods Reverse transcription-poly-merase chain reaction (RT-PCR) and PPARβ/δ luciferase activity experiment to detect PPARβ/δ,PPARγ,phospholipase A2 (PLA2),cytochrome C oxidase 2 (Cox-2),prostaglandin E synthase (PGES),prostacyclin synthase (PGIS) expression level in NSCLC cell lines,NSCLC tissue and normal tissue sam-ple from NSCLC patients.The effect of promoting NSCLC cell lines proliferation and survival mediated by PPARβ/δ was analyzed by siRNA and flow cytometer.The function of PPARβ/δ in regulating the express of Cox-2 and vascular endothelial growth factor (VEGF) gene was confirmed by medicine experiment.Results H441 cells had high levels of PPARβ/δ,Cox-2,cPLA2,PGES and PPARc.H358 and H23 cells had intermediate levels of PPARβ/δ.The PPARβ/δ mRNA level was higher in tumor tissues compared to normal tissues.PPARβ/δ had the function of increasing NSCLC proliferation and survival and low level cPGI2 (activate the PPARβ/δ) could improving NSCLC proliferation and viability,prolonging the S cell cycle and shortening the G1 cell cycle.But there nearly was no impact in A549 cells.Decreasing the cell proliferation and viability and promoting apoptosis when knock-down the PPARβ/δ by siRNA and regulating the Cox-2 and vascular endothelial growth factor (VEGF) expression by PPARβ/δ.There was increasing of Cox-2 and VEGF mRNA during the GW501516 (the ligand of PPARβ/δ) treating NSCLC.But,no change was detected in A549 cells.Conclusion Up-expression in NSCLC compared to normal tissues.There are many function of PPARβ/δ in NSCLC including promoting NSCLC proliferation,survival and viability and affecting the Cox-2,VEGF expression.Together,these data suggest that PPARβ/δ might local the central of multiple signaling pathways and control the NSCLC proliferation.