中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2011年
5期
356-361
,共6页
连渊娥%刘景丰%汪晓军%臧盛兵%黄爱民
連淵娥%劉景豐%汪曉軍%臧盛兵%黃愛民
련연아%류경봉%왕효군%장성병%황애민
癌,肝细胞%肿瘤侵润%肿瘤转移%微粒体前列腺素E合成酶-1
癌,肝細胞%腫瘤侵潤%腫瘤轉移%微粒體前列腺素E閤成酶-1
암,간세포%종류침윤%종류전이%미립체전렬선소E합성매-1
Carcinoma,hepatocellular%Neoplasm invasiveness%Neoplasm metastasis%Microsomal prostaglandin E synthase-1
目的 检测微粒体前列腺素E合成酶(mPGES)-1在肝细胞癌(HCC)组织中的表达,观察其抑制剂MK886下调mPGES-1表达后对肝癌细胞株HepG2生物学行为的影响,探讨mPGES-1在HCC发生、发展及侵袭转移中的作用.方法 收集HCC,癌旁、远癌及正常肝组织标本,用RT-PCR及Western blot检测mPGES-1 mRNA及蛋白的相对表达量.分别采用四甲基偶氮唑盐法、Transwell法检测MK886下调mPGES-1表达后对HepG2细胞增殖,黏附,迁移、侵袭能力的影响.计量资料若数据呈正态分布且方差齐,多组间均数比较用单因素方差分析,组间两两均数比较用LSD-t检验;若数据非正态分布或方差不齐,用多个样本比较的Kruskal-Wallis H秩和检验,两两比较用Mann-Whitney U秩和检验.结果 mPGES-1在HCC中表达水平高于正常肝组织(P<0.01);随病理学分级的升高,mPGES-1表达量逐渐增高;包膜侵犯组中mPGES-1表达量高于无包膜侵犯组(P<0.01);转移组高于无转移组(P<0.01).MK886作用于HepG2细胞48 h后,mPGES-1 mRNA 及蛋白表达水平明显下降(F=140.402,P<0.01; α'=0.00714,P<0.01),并呈剂量依赖性.与对照组比较,MK886作用后HepG2细胞增殖抑制率呈明显的时间和剂量依赖性.20、40,75 μmol/LMK886实验组细胞黏附率分别为85.3%±1.3%、70.5%±1.5%、45.8%±2.4%,明显低于对照组的100.0%±0(F=626.313,P<0.01).迁移实验显示20、30、40 vmol/L MK886实验组细胞24h的迁移细胞数分别为每高倍视野(92.47±1.90)个、(62.63±1.96)个、(37.33±0.83)个,低于对照组迁移细胞数[(128.93±2.60)个,F=1253.805,P<0.01].侵袭实验显示20,30,40μmol/L MK886实验组细胞24 h的侵袭细胞数分别为每高倍视野(41.67±1.30)个、(25.47±1.30)个,(13.93±1.66)个,对照组侵袭细胞数为(55.67±2.08)个,各组间差异均有统计学意义(F=372.615,P<0.01).MK886作用后HepG2细胞的黏附率、迁移及侵袭细胞数均呈剂量依赖性降低.结论 mPGEs-1与HCC的发生、发展密切相关,其表达下调预示可降低HCC的侵袭、转移潜能.
目的 檢測微粒體前列腺素E閤成酶(mPGES)-1在肝細胞癌(HCC)組織中的錶達,觀察其抑製劑MK886下調mPGES-1錶達後對肝癌細胞株HepG2生物學行為的影響,探討mPGES-1在HCC髮生、髮展及侵襲轉移中的作用.方法 收集HCC,癌徬、遠癌及正常肝組織標本,用RT-PCR及Western blot檢測mPGES-1 mRNA及蛋白的相對錶達量.分彆採用四甲基偶氮唑鹽法、Transwell法檢測MK886下調mPGES-1錶達後對HepG2細胞增殖,黏附,遷移、侵襲能力的影響.計量資料若數據呈正態分佈且方差齊,多組間均數比較用單因素方差分析,組間兩兩均數比較用LSD-t檢驗;若數據非正態分佈或方差不齊,用多箇樣本比較的Kruskal-Wallis H秩和檢驗,兩兩比較用Mann-Whitney U秩和檢驗.結果 mPGES-1在HCC中錶達水平高于正常肝組織(P<0.01);隨病理學分級的升高,mPGES-1錶達量逐漸增高;包膜侵犯組中mPGES-1錶達量高于無包膜侵犯組(P<0.01);轉移組高于無轉移組(P<0.01).MK886作用于HepG2細胞48 h後,mPGES-1 mRNA 及蛋白錶達水平明顯下降(F=140.402,P<0.01; α'=0.00714,P<0.01),併呈劑量依賴性.與對照組比較,MK886作用後HepG2細胞增殖抑製率呈明顯的時間和劑量依賴性.20、40,75 μmol/LMK886實驗組細胞黏附率分彆為85.3%±1.3%、70.5%±1.5%、45.8%±2.4%,明顯低于對照組的100.0%±0(F=626.313,P<0.01).遷移實驗顯示20、30、40 vmol/L MK886實驗組細胞24h的遷移細胞數分彆為每高倍視野(92.47±1.90)箇、(62.63±1.96)箇、(37.33±0.83)箇,低于對照組遷移細胞數[(128.93±2.60)箇,F=1253.805,P<0.01].侵襲實驗顯示20,30,40μmol/L MK886實驗組細胞24 h的侵襲細胞數分彆為每高倍視野(41.67±1.30)箇、(25.47±1.30)箇,(13.93±1.66)箇,對照組侵襲細胞數為(55.67±2.08)箇,各組間差異均有統計學意義(F=372.615,P<0.01).MK886作用後HepG2細胞的黏附率、遷移及侵襲細胞數均呈劑量依賴性降低.結論 mPGEs-1與HCC的髮生、髮展密切相關,其錶達下調預示可降低HCC的侵襲、轉移潛能.
목적 검측미립체전렬선소E합성매(mPGES)-1재간세포암(HCC)조직중적표체,관찰기억제제MK886하조mPGES-1표체후대간암세포주HepG2생물학행위적영향,탐토mPGES-1재HCC발생、발전급침습전이중적작용.방법 수집HCC,암방、원암급정상간조직표본,용RT-PCR급Western blot검측mPGES-1 mRNA급단백적상대표체량.분별채용사갑기우담서염법、Transwell법검측MK886하조mPGES-1표체후대HepG2세포증식,점부,천이、침습능력적영향.계량자료약수거정정태분포차방차제,다조간균수비교용단인소방차분석,조간량량균수비교용LSD-t검험;약수거비정태분포혹방차불제,용다개양본비교적Kruskal-Wallis H질화검험,량량비교용Mann-Whitney U질화검험.결과 mPGES-1재HCC중표체수평고우정상간조직(P<0.01);수병이학분급적승고,mPGES-1표체량축점증고;포막침범조중mPGES-1표체량고우무포막침범조(P<0.01);전이조고우무전이조(P<0.01).MK886작용우HepG2세포48 h후,mPGES-1 mRNA 급단백표체수평명현하강(F=140.402,P<0.01; α'=0.00714,P<0.01),병정제량의뢰성.여대조조비교,MK886작용후HepG2세포증식억제솔정명현적시간화제량의뢰성.20、40,75 μmol/LMK886실험조세포점부솔분별위85.3%±1.3%、70.5%±1.5%、45.8%±2.4%,명현저우대조조적100.0%±0(F=626.313,P<0.01).천이실험현시20、30、40 vmol/L MK886실험조세포24h적천이세포수분별위매고배시야(92.47±1.90)개、(62.63±1.96)개、(37.33±0.83)개,저우대조조천이세포수[(128.93±2.60)개,F=1253.805,P<0.01].침습실험현시20,30,40μmol/L MK886실험조세포24 h적침습세포수분별위매고배시야(41.67±1.30)개、(25.47±1.30)개,(13.93±1.66)개,대조조침습세포수위(55.67±2.08)개,각조간차이균유통계학의의(F=372.615,P<0.01).MK886작용후HepG2세포적점부솔、천이급침습세포수균정제량의뢰성강저.결론 mPGEs-1여HCC적발생、발전밀절상관,기표체하조예시가강저HCC적침습、전이잠능.
Objective To study the expression of mPGES-1 in hepatocellular carcinoma(HCC),observe the effect of MK886 on down-regulation of mPGES-1 gene expression on the biology of human hepatocarcinoma cell line HepG2 and to investigate its significance in the occurrence, progression, metastasis and invasion. Methods HCC tissues, para-carcinoma tissues, far-carcinoma tissues and normal liver tissues were collected. The expressions of mPGES-1 were determined by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot. The proliferation, adherence, migration and invasion abilities of HepG2 cells interfered by MK886 were assessed by MTT and transwell technique respectively. Results The expression of mPGES-1 in HCCs was higher than that in normal liver tissues (P < 0.01), which increased following histological grade. Furthermore, mPGES-1 expression level was higher in the capsule invasion and metastasis tumor than in primary locus. A significant dose-dependent down-regulation of expressions of mPGES-1 gene mRNA and protein were observed in HepG2 cells when MK886 was given for 48 h (F = 140.402, P < 0.01;α' = 0.00714, P < 0.01). Compared with the control group, the growth inhibitory rate of HepG2 cell was observed significantly time and dose-dependent when MK886 was given. The rate of adhesion cells in experimental groups were 85.3% ± 1.3%, 70.5% ± 1.5% and 45.8% ± 2.4%, respectively, less than that in control group 100.0% ± 0 (F = 626.313, P < 0.01). The migration cells was 92.47 ± 1.90, 62.63 ± 1.96 and 37.33 + 0.83 respectively in the experimental groups after 24h, lower than that in the control group 128.93 ± 2.60 (F = 1253.805, P < 0.01). The invasion assay revealed that the invading cells were 41.67 ± 1.30,25.47 ± 1.30 and 13.93 ± 1.66 in the experimental groups, in contrast to 55.67 ± 2.08 in control group after 24 h. The differerce between these groups was significant (F = 372.615, P < 0.01). The numbers of adhesion, migration and invasion of HepG2 cells were dose-dependent in MK886 groups. Conclusion Overexpression of mPGES-1 was associated with the tumorigenesis and progression of HCC. The down-regulation of mPGES-1 gene expression might indicated the decrease of the invasion and metastasis of HCC.
