抑制X染色体连锁的凋亡抑制蛋白(XIAP)和Survivin表达对胰腺癌Panc-1细胞增殖及化疗敏感性的影响  

X-linked inhibitor of apoptosis protein (XIAP) and Survivin suppression on human pancreatic cancer cells Panc-1 proliferation and chemosensitivety

作　　者：宰红艳[1] 易小平[2] 李宜雄[1] 龙学颖[2] 曹丽平[1] 刘慧[2] 

机构地区：[1]中南大学湘雅医院普外科,长沙410008 [2]中南大学湘雅医院放射科,长沙410008

出　　处：《北京大学学报（医学版）》2013年第2期242-249,共8页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(30872492);湖南省自然科学基金(08JJ3042);中南大学自由探索计划(2011QNZT153)资助~~

摘　　要：目的:探讨同时抑制X染色体连锁的凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)和Sur-vivin表达,对胰腺癌Panc-1细胞增殖及吉西他滨(Gem)化疗敏感性的影响,并与单独抑制XIAP或Survivin表达的策略进行对比。方法:运用前期实验构建的XIAP-shRNA慢病毒(LV-X)和Survivin-shRNA慢病毒(LV-S),分别建立XIAP和/或Survivin表达稳定抑制的胰腺癌Panc-1细胞株,即Panc-1-X、Panc-1-S和Panc-1-XS。运用Real-timePCR和半定量Western blot分别检测XIAP和Survivin的mRNA和蛋白的表达情况,以细胞计数法及克隆形成实验检测细胞增殖能力,Caspase-3/7试剂盒及流式细胞仪检测细胞凋亡,MTT法检测细胞对Gem的化疗敏感性。结果:成功建立了XIAP和/或Survivin表达稳定抑制的胰腺癌细胞株Panc-1。XIAP和Survivin同时稳定抑制后,Panc-1的增殖能力显著受到抑制,Panc-1-XS组的克隆形成率为10.12%±1.33%,显著低于对照组Panc-1-XncSnc组(96.61%±7.89%)和Panc-1组(100.28%±8.97%,P<0.05)。用0.5 mg/L Gem处理24 h后,Panc-1-XS组的Caspase-3/7相对活性明显升高至15.02±0.57,显著高于Panc-1组与Panc-1-XncSnc组(分别为8.87±0.19和9.05±0.23,P<0.05);Panc-1-XS组的细胞凋亡率为24.09%±2.75%,显著高于对照组Panc-1-XncSnc及Panc-1组(分别为12.09%±1.97%和12.06%±1.22%,P<0.05)。Panc-1-XS组的IC50值为(0.47±0.04)mg/L,显著低于对照组Panc-1-XncSnc的(2.18±0.13)mg/L及Panc-1组的(2.13±0.18)mg/L(P<0.05),对Gem的化疗敏感性显著增强。进一步检测显示,Panc-1-XS组的IC50值为(0.47±0.04)mg/L,均显著低于Panc-1-X组的(0.76±0.07)mg/L和Panc-1-S组的(0.87±0.09)mg/L(P<0.05)。结论:同时稳定抑制胰腺癌细胞株Panc-1中XIAP和Survivin的表达,能显著抑制Panc-1细胞增殖能力,增强其Gem化疗敏感性,并明显优于XIAP或Survivin表达单独抑制的策略。Objective：To investigate the effect on cell proliferation and chemosensitivity of human pan-creatic cancer cells Panc-1 after X-linked inhibitor of apoptosis protein （XIAP） and Survivin are inhibi-ted simultaneously, and to compare it with the separate gene suppression strategy by which expression of XIAP or Survivin is inhibited respectively. Methods： Panc-1 （Panc-l-X,Panc-l-S and Panc-l-XS） in which expression of XIAP and/or Survivin was inhibited, was established by using XIAP-shRNA lentiviral and Survivin-shRNA lentiviral we had buih. The expressions of XIAP and Survivin mRNA and protein were evaluated by Real-time PCR and Semi-quantitatively Western blot analysis; cell proliferation was in-vestigated by cell counting and colony formation assay; cell apoptosis was investigated by Caspase-3/7 ac-tivity assay kit and flow cytometry; gemcitabine （Gem） chemosensitivity was investigated by MTT assay. Results： The pancreatic cell line Panc-1 in which the expression of XIAP and/or Survivin was stablely inhibited was successfully established. The cell proliferation of Panc-l-XS cells decreased significantly. The colony formation rate of Panc-l-XS cells （ 10.12% ± 1.33% ）, was significantly lower than that of Panc-l-XncSnc cells （96.61% ±7.89% ） and Panc-1 cells （ 100.28% ± 8.97% ） respectively （P 〈 0, 05 ）. After being treated by 0.5 mg/L Gem for 24 h, the Caspase-3/7 relative activity of Panc-l-XS cells （ 15.02 ± 0.57 ） was significantly higher than that of Panc-1 cells and Panc-l-XncSnc cells （8.87 ±0.19 and 9.05 ±0.23, respectively; P 〈 0.05）; and the rate of apoptosis of Panc-l-XS cells （24.09% ± 2. 75% ） was significantly higher than that of Panc-l-XncSnc cells and Panc-1 ceils （12,09% ±1.97% and 12.06% ± 1, 22%, respectively; P 〈0.05）. The IC50 value of Panc-l-XS cells [ （0.47 ± 0.04） mg/L ] was significantly lower than that of Panc-1-XncSnc cells [ （2.18 -± 0.13 ） mg/L] and Panc-1 cells [ （2.13 ±0.18） rag/L, P 〈0.05]. Further tes

关 键 词：胰腺肿瘤 凋亡抑制蛋白质类 细胞增殖 药物疗法 抗药性 肿瘤 

分 类 号：R736.7[医药卫生—肿瘤]