中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
11期
2345-2348
,共4页
笪良山%张帅%尹荣%许磊%钱亦淳%蒋峰%许林
笪良山%張帥%尹榮%許磊%錢亦淳%蔣峰%許林
달량산%장수%윤영%허뢰%전역순%장봉%허림
塞来昔布%手术应激%前列腺素E2%肺转移%机制
塞來昔佈%手術應激%前列腺素E2%肺轉移%機製
새래석포%수술응격%전렬선소E2%폐전이%궤제
Celecoxib%Surgery stress%Prostaglandin E2%Lung metastasis%Mechanism
目的 探讨塞来昔布(Celecoxib)对肺腺癌手术应激所致肺转移的影响及其分子作用机制.方法 利用BALB/c裸鼠,通过开胸手术同时尾静脉注射肺腺癌细胞株(A549)建立手术应激所致肺转移动物模型.将75只裸鼠随机分成3组:对照组、手术组、手术+Celecoxib组.3组小鼠均尾静脉注射A549细胞1×106个,4周后处死所有小鼠,5％苦味酸染色比较各组小鼠肺表面结节数；苏木素-伊红(HE)染色明确各组肺结节病理性质,酶联免疫吸附试验(ELISA)试剂盒检测不同时间点小鼠血清前列腺素E2(PGE2)水平.利用A549细胞进行划痕实验、Transwell小室实验检测体外PGE2和Celecoxib对A549迁移和侵袭能力的影响,实时定量聚合酶链反应(Real-time PCR)和Western blot检测PGE2和Celecoxib对细胞基质金属蛋白酶(MMP)-2/-9、E-钙黏蛋白(E-cadherin)、糖原合成激酶-3β(GSK-3β)和细胞核β-连环蛋白(β-catenin)表达的影响.结果 HE染色确定小鼠肺表面结节系A549细胞肺转移灶.与对照组比较,手术组小鼠肺表面肺结节数明显增多[(30.2±8.8)个比(17.2±5.2)个,P＜0.05],术后第3天血清平均PGE2水平较术前明显升高达到(212.8±52.9)ng/L,而手术组给予Celecoxib则能明显抑制肺结节数增多[(9.1±4.2)个]和血清PGE2升高(102.7±11.2) ng/L,差异有统计学意义(P＜0.05).细胞划痕和Transwell实验也显示Celecoxib能明显抑制PGE2对A549的促迁移和侵袭作用[(79.8±9.9)％比(177.8±13.2)％,P＜0.01].Western blot结果与Real-time PCR结果相似:PGE2组细胞MMP-9 mRNA和蛋白表达增加,E-cadherin表达明显下降,而经Celecoxib处理后PGE2对上述基因表达的影响被逆转.Western blot结果显示,Celecoxib能抑制PGE2刺激引起的GSK-3β活化和细胞核β-catenin表达量升高.结论 Celecoxib能明显抑制肺腺癌手术应激所致肺转移,其分子机制可能与抑制β-catenin核转位有关.
目的 探討塞來昔佈(Celecoxib)對肺腺癌手術應激所緻肺轉移的影響及其分子作用機製.方法 利用BALB/c裸鼠,通過開胸手術同時尾靜脈註射肺腺癌細胞株(A549)建立手術應激所緻肺轉移動物模型.將75隻裸鼠隨機分成3組:對照組、手術組、手術+Celecoxib組.3組小鼠均尾靜脈註射A549細胞1×106箇,4週後處死所有小鼠,5％苦味痠染色比較各組小鼠肺錶麵結節數；囌木素-伊紅(HE)染色明確各組肺結節病理性質,酶聯免疫吸附試驗(ELISA)試劑盒檢測不同時間點小鼠血清前列腺素E2(PGE2)水平.利用A549細胞進行劃痕實驗、Transwell小室實驗檢測體外PGE2和Celecoxib對A549遷移和侵襲能力的影響,實時定量聚閤酶鏈反應(Real-time PCR)和Western blot檢測PGE2和Celecoxib對細胞基質金屬蛋白酶(MMP)-2/-9、E-鈣黏蛋白(E-cadherin)、糖原閤成激酶-3β(GSK-3β)和細胞覈β-連環蛋白(β-catenin)錶達的影響.結果 HE染色確定小鼠肺錶麵結節繫A549細胞肺轉移竈.與對照組比較,手術組小鼠肺錶麵肺結節數明顯增多[(30.2±8.8)箇比(17.2±5.2)箇,P＜0.05],術後第3天血清平均PGE2水平較術前明顯升高達到(212.8±52.9)ng/L,而手術組給予Celecoxib則能明顯抑製肺結節數增多[(9.1±4.2)箇]和血清PGE2升高(102.7±11.2) ng/L,差異有統計學意義(P＜0.05).細胞劃痕和Transwell實驗也顯示Celecoxib能明顯抑製PGE2對A549的促遷移和侵襲作用[(79.8±9.9)％比(177.8±13.2)％,P＜0.01].Western blot結果與Real-time PCR結果相似:PGE2組細胞MMP-9 mRNA和蛋白錶達增加,E-cadherin錶達明顯下降,而經Celecoxib處理後PGE2對上述基因錶達的影響被逆轉.Western blot結果顯示,Celecoxib能抑製PGE2刺激引起的GSK-3β活化和細胞覈β-catenin錶達量升高.結論 Celecoxib能明顯抑製肺腺癌手術應激所緻肺轉移,其分子機製可能與抑製β-catenin覈轉位有關.
목적 탐토새래석포(Celecoxib)대폐선암수술응격소치폐전이적영향급기분자작용궤제.방법 이용BALB/c라서,통과개흉수술동시미정맥주사폐선암세포주(A549)건립수술응격소치폐전이동물모형.장75지라서수궤분성3조:대조조、수술조、수술+Celecoxib조.3조소서균미정맥주사A549세포1×106개,4주후처사소유소서,5％고미산염색비교각조소서폐표면결절수；소목소-이홍(HE)염색명학각조폐결절병이성질,매련면역흡부시험(ELISA)시제합검측불동시간점소서혈청전렬선소E2(PGE2)수평.이용A549세포진행화흔실험、Transwell소실실험검측체외PGE2화Celecoxib대A549천이화침습능력적영향,실시정량취합매련반응(Real-time PCR)화Western blot검측PGE2화Celecoxib대세포기질금속단백매(MMP)-2/-9、E-개점단백(E-cadherin)、당원합성격매-3β(GSK-3β)화세포핵β-련배단백(β-catenin)표체적영향.결과 HE염색학정소서폐표면결절계A549세포폐전이조.여대조조비교,수술조소서폐표면폐결절수명현증다[(30.2±8.8)개비(17.2±5.2)개,P＜0.05],술후제3천혈청평균PGE2수평교술전명현승고체도(212.8±52.9)ng/L,이수술조급여Celecoxib칙능명현억제폐결절수증다[(9.1±4.2)개]화혈청PGE2승고(102.7±11.2) ng/L,차이유통계학의의(P＜0.05).세포화흔화Transwell실험야현시Celecoxib능명현억제PGE2대A549적촉천이화침습작용[(79.8±9.9)％비(177.8±13.2)％,P＜0.01].Western blot결과여Real-time PCR결과상사:PGE2조세포MMP-9 mRNA화단백표체증가,E-cadherin표체명현하강,이경Celecoxib처리후PGE2대상술기인표체적영향피역전.Western blot결과현시,Celecoxib능억제PGE2자격인기적GSK-3β활화화세포핵β-catenin표체량승고.결론 Celecoxib능명현억제폐선암수술응격소치폐전이,기분자궤제가능여억제β-catenin핵전위유관.
Objective To explore the effects of Celecoxib on lung metastasis of lung adeno-carcinoma initiated by thoracotomy stress and the underlying mechanism.Methods Animal models of metastasis of lung adeno-carcinoma initiated by thoracotomy stress were created by thoracotomy and injection of lung adeno-carcinoma cell line (A549) via the lateral tail vein into BALB/c nude mice.Seventy-five nude mices were randomly divided into 3 groups:control,mice receiving thoracotomy; mice receiving thoracotomy and Celecoxib.All nude mices were injected with a suspension of 1 x 106 A549 cells via the lateral tail vein and killed at 4th week.The nodules on the lung surface were stained with 5％ picric acid and the number of nodules in each group was analyzed.In addition,the nodules were also cut into sections for hematoxylin and eosin (HE) staining to identify their pathological features.The serum prostaglandin E2 (PGE2) level was determined using the PGE2 enzyme linked immunosorbent assay (ELISA) kit at different time points.The effects of Celecoxib and PGE2 on cell migration and invasion were studied by wound healing and Transwell assay.Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were applied to detect the expression of matrix metalloproteinase (MMP)-2/-9,E-cadherin,GSK-3β and β-catenin in A549 cells.Results HE staining showed that the metastatic nodules of A549 cells on the lung surface were identified.The number of metastasis in mice receiving thoracotomy was greater than that in control mice (30.2 ±8.8 vs.17.2 ±5.2,P ＜0.05) and serum PGE2 level in mice receiving thoraeotomy was increased to (212.8 ±52.9) ng/L at 31st day after surgery.However,Celecoxib inhibited the increased nodules of metastases (9.1 ± 4.2) and serum PGE2 level [(102.7 ± 11.2) ng/L] respectively (P ＜0.05).PGE2 increased migration and invasion of A549 cells in the wound healing and Transwell assays,which could be reversed by Celecoxib [(177.8 ± 13.2) ％ vs.(79.8 ± 9.9) ％,P ＜ 0.01].Real-time PCR showed that mRNA expression of MMP-9 was increased by PGE. and reversed by Celecoxib,while E-cadherin was markedly decreased,and Celecoxib increased its expression.The results of Western blotting showed similar effects in the protein expression of MMP-9 and E-cadherin.Western blotting also showed that the effects of PGE2-induced increase in the protein level of β-catenin in the nucleus of A549 cells and activation of GSK-3β were significantly inhibited by Celecoxib.Conclusion Celecoxib markedly inhibits lung metastasis of lung adeno-carcinoma initiated by thoracotomy stress and the underlying mechanism may associate with inhibiting translocation of β-catenin into nucleus.
