肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2011年
7期
433-437
,共5页
李金凤%刘春英%邓卫巍%刘春晖
李金鳳%劉春英%鄧衛巍%劉春暉
리금봉%류춘영%산위외%류춘휘
肿瘤,实验性%结肠肿瘤%药物疗法,联合%肿瘤转移
腫瘤,實驗性%結腸腫瘤%藥物療法,聯閤%腫瘤轉移
종류,실험성%결장종류%약물요법,연합%종류전이
Neoplasms,experimental%Colonic neoplasms%Drug therapy,combination%Neoplasms metastasis
目的 对比研究单独及联合应用参与花生四烯酸代谢的环氧合酶-2(COX-2)及5-脂氧合酶(5-LOX)抑制剂塞来昔布与多两苯醌(AA861)代谢途径对结肠癌细胞转移的抑制作用.方法 体外分组培养具有高转移能力的人类结肠癌HT-29细胞,药物分别作用48 h后,采用四甲基偶氮唑盐比色(MTT)法榆测AA861及塞来昔布对结肠癌细胞增殖的影响,Transwell法检测不同药物分组对结肠癌HT-29细胞迁移能力的影响,免疫荧光染色法检测细胞内细胞问黏附分子-1(ICAM-1)蛋白的变化,反转录-聚合酶链(RT-PCR)法检测细胞内ICAM-1及血管内皮生长因子(VEGF)基因表达变化.结果 AA861与塞来昔布单独应用都能以时间浓度依赖性抑制结肠癌细胞增殖与迁移,其中AA861与塞莱昔布100μmol/L分别单独应用时对结肠癌细胞增殖的抑制率分别为43.2%、42.8%,两者100μmol/L联合应用时对结肠癌细胞增殖的抑制率为53.8%,与分别单独应用时比较差异有统计学意义(均P＜0.001);AA861与塞莱昔布100 μmol/L分别单独应用时对结肠癌细胞迁移的抑制率为32.0%、29.3%,两者100μmol/L联合应用时对结肠癌细胞增殖的抑制率为57.8%,与分别单独应用时比较差异有统计学意义(均P＜0.001).免疫荧光染色及RT-PCR提示两种药物都能抑制结肠癌细胞内VEGF与ICAM-1蛋白及基因表达,小剂量联合应用都能抑制结肠癌细胞内VEGF与ICAM-1蛋白及基因表达,且两者联合应用比单独应用抑制作用强(P＜0.001).结论 小剂量塞来昔布及AA861联合应用能以协同作用抑制结肠癌细胞的增殖与迁移,其机制可能与抑制细胞内肿瘤转移相关因子VEGF及ICAM-1表达有关.
目的 對比研究單獨及聯閤應用參與花生四烯痠代謝的環氧閤酶-2(COX-2)及5-脂氧閤酶(5-LOX)抑製劑塞來昔佈與多兩苯醌(AA861)代謝途徑對結腸癌細胞轉移的抑製作用.方法 體外分組培養具有高轉移能力的人類結腸癌HT-29細胞,藥物分彆作用48 h後,採用四甲基偶氮唑鹽比色(MTT)法榆測AA861及塞來昔佈對結腸癌細胞增殖的影響,Transwell法檢測不同藥物分組對結腸癌HT-29細胞遷移能力的影響,免疫熒光染色法檢測細胞內細胞問黏附分子-1(ICAM-1)蛋白的變化,反轉錄-聚閤酶鏈(RT-PCR)法檢測細胞內ICAM-1及血管內皮生長因子(VEGF)基因錶達變化.結果 AA861與塞來昔佈單獨應用都能以時間濃度依賴性抑製結腸癌細胞增殖與遷移,其中AA861與塞萊昔佈100μmol/L分彆單獨應用時對結腸癌細胞增殖的抑製率分彆為43.2%、42.8%,兩者100μmol/L聯閤應用時對結腸癌細胞增殖的抑製率為53.8%,與分彆單獨應用時比較差異有統計學意義(均P＜0.001);AA861與塞萊昔佈100 μmol/L分彆單獨應用時對結腸癌細胞遷移的抑製率為32.0%、29.3%,兩者100μmol/L聯閤應用時對結腸癌細胞增殖的抑製率為57.8%,與分彆單獨應用時比較差異有統計學意義(均P＜0.001).免疫熒光染色及RT-PCR提示兩種藥物都能抑製結腸癌細胞內VEGF與ICAM-1蛋白及基因錶達,小劑量聯閤應用都能抑製結腸癌細胞內VEGF與ICAM-1蛋白及基因錶達,且兩者聯閤應用比單獨應用抑製作用彊(P＜0.001).結論 小劑量塞來昔佈及AA861聯閤應用能以協同作用抑製結腸癌細胞的增殖與遷移,其機製可能與抑製細胞內腫瘤轉移相關因子VEGF及ICAM-1錶達有關.
목적 대비연구단독급연합응용삼여화생사희산대사적배양합매-2(COX-2)급5-지양합매(5-LOX)억제제새래석포여다량분곤(AA861)대사도경대결장암세포전이적억제작용.방법 체외분조배양구유고전이능력적인류결장암HT-29세포,약물분별작용48 h후,채용사갑기우담서염비색(MTT)법유측AA861급새래석포대결장암세포증식적영향,Transwell법검측불동약물분조대결장암HT-29세포천이능력적영향,면역형광염색법검측세포내세포문점부분자-1(ICAM-1)단백적변화,반전록-취합매련(RT-PCR)법검측세포내ICAM-1급혈관내피생장인자(VEGF)기인표체변화.결과 AA861여새래석포단독응용도능이시간농도의뢰성억제결장암세포증식여천이,기중AA861여새래석포100μmol/L분별단독응용시대결장암세포증식적억제솔분별위43.2%、42.8%,량자100μmol/L연합응용시대결장암세포증식적억제솔위53.8%,여분별단독응용시비교차이유통계학의의(균P＜0.001);AA861여새래석포100 μmol/L분별단독응용시대결장암세포천이적억제솔위32.0%、29.3%,량자100μmol/L연합응용시대결장암세포증식적억제솔위57.8%,여분별단독응용시비교차이유통계학의의(균P＜0.001).면역형광염색급RT-PCR제시량충약물도능억제결장암세포내VEGF여ICAM-1단백급기인표체,소제량연합응용도능억제결장암세포내VEGF여ICAM-1단백급기인표체,차량자연합응용비단독응용억제작용강(P＜0.001).결론 소제량새래석포급AA861연합응용능이협동작용억제결장암세포적증식여천이,기궤제가능여억제세포내종류전이상관인자VEGF급ICAM-1표체유관.
Objective To investigate the effects of cyclooxygenase-2 (COX-2) selective inhibitor combined with 5-lipoxygenase (5-LOX) inhibitor docebenone (AA861) on cell proliferation and migration of human colon cancer cell line HT-29. Methods Cultured in vitro of HT-29 cells in high metastatic colon cancer, A A861 and celecoxib on colon cancer cell drugs for 48 h, the cell proliferation was assayed by MTT; the migration of cell was detected by Transwell; immunofluorescence staining of intracellular changes in ICAM-1 protein, RT-PCR detect the gene expression of ICAM-1 and VEGF. Results MTT and Transwell experiments showed that the inhibition on celecoxib and AA861 on colon cancer was dose-dependent and time-dependent. And the inhibition of AA861 and celecoxib 100 μmol/L alone on colon cancer cells proliferation were 43.2 % and 42.8%, and when the two drugs 100 μmol/L combined on colon cancer cells the inhibition was 53.8%, and the difference between alone group and combined group was statistical (P ＜0.001). The inhibition of AA861 and celecoxib 100 μmol/L alone on colon cancer cells migration were 32.0 % and 29.3%, and when the two drugs 100 μ,mol/L combined on colon cancer cells the inhibition is 57.8 %, and the difference of between alone group and combined group was statistical (P ＜0.001). Immunefluorescence staining and RT-PCR results suggested that the two drugs can inhibit ICAM-1 and VEGF protein and gene expression, and when the two drugs combined, a stronger inhibition effect appeared than used alone (P＜0.001). Conclusion Low-dose celecoxib and AA861 combined has a synergistic inhibited effect on colon cancer cells invasion and metastasis, and the mechanism relates with the VEGF and ICAM-1 expression.
