中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2013年
3期
211-214
,共4页
郑衍玲%李艳%罗红敏%高杰%田禾%牛作兴%李胜
鄭衍玲%李豔%囉紅敏%高傑%田禾%牛作興%李勝
정연령%리염%라홍민%고걸%전화%우작흥%리성
胰腺肿瘤%TNF相关凋亡诱导配体%细胞凋亡%吉西他滨
胰腺腫瘤%TNF相關凋亡誘導配體%細胞凋亡%吉西他濱
이선종류%TNF상관조망유도배체%세포조망%길서타빈
Pancreatic neoplasms%TNF related apoptosis-inducing ligand%Apoptosis%Gemcitabine
目的 探讨外源性Smac/DIABLO对人胰腺癌SW1990细胞生物学特性和对TRAIL及吉西他滨化疗敏感性的影响. 方法 利用脂质体2000介导Smac/DIABLO基因转染胰腺癌SW1990细胞获得细胞SW1990/Smac,转染空载体为对照组(SW1990/neo)；在不同浓度和时间下以肿瘤坏死因子相关凋亡诱导配体(TRAIL)和吉西他滨处理2组细胞株并分为TRAIL组、吉西他滨组和联合组.MTT法检测细胞株的生长抑制率,流式细胞仪检测细胞凋亡率及凋亡细胞形态,Western blot检测凋亡相关蛋白Smac/DIABLO、抑制凋亡蛋白XIAP、细胞色素C及细胞凋亡因子caspase-3的表达.结果 转染Smac/DIABLO的细胞生长明显落后于转染空载体的细胞.TRAIL浓度为200、500、1000、2500 ng/ml时,作用24 h对SW1990/neo和SW1990/Smac细胞的抑制率分别为11.11％、46.03％、67.08％、76.19％及22.11％、42.67％、56.63％、67.6％ (P ＜0.05).吉西他滨浓度分别为10、20、40、60μmol/L作用24 h对SW1990/neo和SW1990/Smac的抑制率分别为15.2％、34.6％、55.16％、76.4％和22.65％、36.85％、55.11％、79.99％ (P＜0.05).以TRAIL(500 ng/ml)、吉西他滨(20 μmol/L)及TRAIL(500 ng/ml)+吉西他滨(20 μmol/L)作用24 h后,SW1990/neo和SW1990/Smac细胞的凋亡率分别为5.64％、15.30％、27.27％和20.37％、23.27％、67.30％ (P ＜0.05).SW1990/Smac细胞在TRAIL及吉西他滨作用后,细胞内促凋亡蛋白Smac/DIABLO、细胞色素C及caspase-3活化片段表达均显著升高,而抑制凋亡蛋白XIAP表达显著降低(P＜0.05).结论 Smac/DIABLO可诱导SW1990细胞的凋亡、抑制细胞增殖,并增强胰腺癌细胞对TRAIL及吉西他滨的化疗敏感性,其机制可能与Smac/DIABLO、细胞色素C、XIAP及caspase-3的活性有关.
目的 探討外源性Smac/DIABLO對人胰腺癌SW1990細胞生物學特性和對TRAIL及吉西他濱化療敏感性的影響. 方法 利用脂質體2000介導Smac/DIABLO基因轉染胰腺癌SW1990細胞穫得細胞SW1990/Smac,轉染空載體為對照組(SW1990/neo)；在不同濃度和時間下以腫瘤壞死因子相關凋亡誘導配體(TRAIL)和吉西他濱處理2組細胞株併分為TRAIL組、吉西他濱組和聯閤組.MTT法檢測細胞株的生長抑製率,流式細胞儀檢測細胞凋亡率及凋亡細胞形態,Western blot檢測凋亡相關蛋白Smac/DIABLO、抑製凋亡蛋白XIAP、細胞色素C及細胞凋亡因子caspase-3的錶達.結果 轉染Smac/DIABLO的細胞生長明顯落後于轉染空載體的細胞.TRAIL濃度為200、500、1000、2500 ng/ml時,作用24 h對SW1990/neo和SW1990/Smac細胞的抑製率分彆為11.11％、46.03％、67.08％、76.19％及22.11％、42.67％、56.63％、67.6％ (P ＜0.05).吉西他濱濃度分彆為10、20、40、60μmol/L作用24 h對SW1990/neo和SW1990/Smac的抑製率分彆為15.2％、34.6％、55.16％、76.4％和22.65％、36.85％、55.11％、79.99％ (P＜0.05).以TRAIL(500 ng/ml)、吉西他濱(20 μmol/L)及TRAIL(500 ng/ml)+吉西他濱(20 μmol/L)作用24 h後,SW1990/neo和SW1990/Smac細胞的凋亡率分彆為5.64％、15.30％、27.27％和20.37％、23.27％、67.30％ (P ＜0.05).SW1990/Smac細胞在TRAIL及吉西他濱作用後,細胞內促凋亡蛋白Smac/DIABLO、細胞色素C及caspase-3活化片段錶達均顯著升高,而抑製凋亡蛋白XIAP錶達顯著降低(P＜0.05).結論 Smac/DIABLO可誘導SW1990細胞的凋亡、抑製細胞增殖,併增彊胰腺癌細胞對TRAIL及吉西他濱的化療敏感性,其機製可能與Smac/DIABLO、細胞色素C、XIAP及caspase-3的活性有關.
목적 탐토외원성Smac/DIABLO대인이선암SW1990세포생물학특성화대TRAIL급길서타빈화료민감성적영향. 방법 이용지질체2000개도Smac/DIABLO기인전염이선암SW1990세포획득세포SW1990/Smac,전염공재체위대조조(SW1990/neo)；재불동농도화시간하이종류배사인자상관조망유도배체(TRAIL)화길서타빈처리2조세포주병분위TRAIL조、길서타빈조화연합조.MTT법검측세포주적생장억제솔,류식세포의검측세포조망솔급조망세포형태,Western blot검측조망상관단백Smac/DIABLO、억제조망단백XIAP、세포색소C급세포조망인자caspase-3적표체.결과 전염Smac/DIABLO적세포생장명현락후우전염공재체적세포.TRAIL농도위200、500、1000、2500 ng/ml시,작용24 h대SW1990/neo화SW1990/Smac세포적억제솔분별위11.11％、46.03％、67.08％、76.19％급22.11％、42.67％、56.63％、67.6％ (P ＜0.05).길서타빈농도분별위10、20、40、60μmol/L작용24 h대SW1990/neo화SW1990/Smac적억제솔분별위15.2％、34.6％、55.16％、76.4％화22.65％、36.85％、55.11％、79.99％ (P＜0.05).이TRAIL(500 ng/ml)、길서타빈(20 μmol/L)급TRAIL(500 ng/ml)+길서타빈(20 μmol/L)작용24 h후,SW1990/neo화SW1990/Smac세포적조망솔분별위5.64％、15.30％、27.27％화20.37％、23.27％、67.30％ (P ＜0.05).SW1990/Smac세포재TRAIL급길서타빈작용후,세포내촉조망단백Smac/DIABLO、세포색소C급caspase-3활화편단표체균현저승고,이억제조망단백XIAP표체현저강저(P＜0.05).결론 Smac/DIABLO가유도SW1990세포적조망、억제세포증식,병증강이선암세포대TRAIL급길서타빈적화료민감성,기궤제가능여Smac/DIABLO、세포색소C、XIAP급caspase-3적활성유관.
Objective To explore the effect of ectopic overexpression of Smac/DIABLO on the proliferation of pancreatic cancer cell line SW1990,and the sensitization to TRAIL and Gemcitabine induced apoptosis.Methods The Smac/DIABLO gene was transfected into the pancreatic cancer cell line SW1990 with the participation of Lipofectamine 2000 (SW1990/Smac).The cell line transfected with empty vector served as controls (SW1990/neo).The SW1990/neo and SW1990/Smac cells were assigned into the following treatment groups:TRAIL group,Gemcitabine group,TRAIL plus Gemcitabine group,and the control group.The SW1990 cells were treated with TRAIL and Gemcitabine in different concentrations and time.The cell growth inhibition rate (CGIR) was detected by MTT,the rate of apoptosis was measured by flow eytometry,the apoptosis morphous was observed by Heochst 33342 staining.The expressions of apoptosis-associated proteins such as Smas/DIABLO,XIAP,cytochrome C and caspase-3 were detected by Western blot.Results The cell growth of SW1990/Smac was significantly lower than growth of SW1990/ neo.The concentration of TRAIL were 200,500,1000 and 2500 ng/ml respectively.After 24 hours,the CGIR of SW1990/neo and SW1990/Smac were 11.11％,46.03％,67.08％,76.19％ and 22.11％,42.67％,56.63％,67.6％ respectively (P ＜ 0.05).The concentration of Gemcitabine were 10,20,40 and 60 μmol/L respectively.After 24 hours,the CGIR of SW1990/neo and SW1990/Smac were 15.2％,34.6％,55.16％,76.4％ and 22.65％,36.85％,55.11％,79.99％ respectively (P＜0.05).The cells of SW1990/neo and SW1990/Smac were treated by TRAIL(500 ng/ml),Gemcitabine (20 μmol/L) and combination group.The apoptosis rate were 5.64％,15.30％,27.27％ and 20.37％,23.27％,67.30％ (P ＜ 0.05) respectively.In combination group,the expressions of activators of caspase such as Smas/DIABLO,cytochrome C and caspase-3 increased significantly,while the expressions of inhibitor of apoptosis protein XIAP decreased.Conclusions Ectopic expression of Smac/DIABLO could induce the apoptosis of SW1990 cell,inhibit the cell proliferation,and enhence the sensitivity of SW1990 cell to TRAIL and Gemcitabine.The mechanism of apoptosis sensitization effect by Smac/DIABLO was associated with significant up-regulation of Smac/DIABLO,cytochrome C,down-regulation of XIAP,and the activation of caspase-3.
