摘要
目的:探讨COX-2抑制剂和survivin反义寡核苷酸联合应用对胰腺癌BxPC-3细胞的抗肿瘤效应及其可能机制.方法:应用胰腺癌BxPC-3细胞进行研究,将BxPC-3细胞分为4组:A组(对照组),B组(Celecoxib 80μmol/L)组,C组(300 nmol/L survivin ASODN),D组(80μmol/L celecoxib+300 nmol/L survivin ASODN).采用MTT检测细胞增殖.流式细胞仪检测细胞凋亡率,caspase-3试剂盒检测caspase-3活性,并用RT-PCR检测Bcl-2、survivin和Mcl-1的mRNA国的变化.结果:将80μmol/L celecoxib和300 nmol/L survivin反义寡核苷酸单独或联合作用于胰腺癌BxPC-3细胞24h和48h,D组细胞的存活率明显低于B,C组(24h:41.0%±0.4% vs 71.0%±2.2%,63.3%±4.5%;48h:34.2%±1.1% vs 61.6%±1.7%,55.0%±3%;P<0.01).作用24h后,流式细胞仪检测细胞凋亡率显示,D组细胞的凋亡率明显高于B,C组(30.33%±3.49% vs 11.93%±1.17%,22.07%±0.93%;P<0.01).caspase-3活性在B,C,D组明显高于对照组(0.04867±0.0021,0.02967±0.0021,0.08767±0.0042 vs 0.007±0.0001;P<0.01),D组细胞的caspase-3活性明显高于B,C组(0.08767±0.0042 vs 0.04867±0.0021,0.02967±0.0021;P<0.01).用100μmol/L塞来昔布作用于胰腺癌BxPC-3细胞24h后,survivin/β-actin和Mcl-1/β-actin的mRNA的比值明显低于对照组(0.68±0.05 vs 1.05±0.06,P<0.01),而Bcl-2/β-actin的mRNA的比值无明显变化(0.99±0.02 vs 1.07±0.06,P>0.05).结论:联合应用COX-2抑制剂塞来昔布和survivin反义寡核苷酸可明显诱导胰腺癌细胞的凋亡,抑制细胞增殖,并能够明显提高caspase-3活性.COX-2抑制剂塞来昔布诱导细胞凋亡可能通过survivin和Mcl-1途径,而非Bcl-2途径.
AIM： To investigate the effects of proliferation and apoptosis in pancreatic cancer cells treated by COX-2 inhibitor celecoxib in combination with survivin antisense oligonucleotides （ASODNs） and their possible mechanism of action. METHODS： This study was carried out on the human pancreatic cancer BxPC-3 cell line. Cellswere divided into four groups： A （control）, B （celecoxib 80 μmol/L）, C （survivin ASODN300 nmol/L）, and D （80 μmol/L celecoxib ＋ 300 nmol/L survivin ASODN）. Cell proliferation was detected with the MTT colorimetric assay, the apoptosis rate in cells was measured by flow cytometry, and caspase-3 activity was evaluated using a caspase-3 assay kit. Using （Reverse Transcription Polymerase Chain Reaction） RTPCR, we examined mRNA for Bcl-2, survivin and Mcl-1. RESULTS： BxPC-3 cells were treated with celecoxib （80 μmol/L） and survivin ASODN （300 nmol/L） alone or in combination for 24 and 48 hours. Cell viability in group D was significantly lower than that in group B or C （24 hours, 41.0% ± 0.4% vs 71.0% ± 2.2% and 63.3% ± 4.5%; 48 hours, 34.2% ± 1.1% vs 61.6% ± 1.7% and 55.0% ± 3%; P 〈 0.01）. After 24 hours, apoptosis mea- sured by flow cytometry in group D was signifi- cantly higher than that in groups B or C （30.33% ± 3.49% vs 11.93% ± 1.17% and 22.07% ± 0.93%; P 〈 0.01）. The caspase-3 activity was significantly increased in group B, C and D compared with the control group （0.04867 ± 0.0021, 0.02967 ± 0.0021 and 0.08767 ± 0.0042 vs 0.007 ± 0.0001; P 〈 0.01）. Caspase-3 activity in group D was signifi- cantly higher than that in groups B and C （0.08767 ± 0.0042 vs 0.04867 ± 0.0021, 0.02967 ± 0.0021; P 〈 0.01）. BxPC-3 cells were treated with celecoxib （100 μmol/L） for 24 hours. The grey ratio of mRNA amplified products for survivin/β-actin and Mcl-1/β-actin was significantly lower in the experimental groups than in the control group （0.68 ± 0.05 vs 1.05 ± 0.06; P 〈 0.01）. However, Bcl-2/β-actin in the expe
出处
《世界华人消化杂志》
CAS
北大核心
2007年第30期3178-3183,共6页
World Chinese Journal of Digestology
