下调自噬性相关蛋白表达对胰腺癌PANC-1细胞增殖的影响  

Influences of pancreatic cancer cell PANC-1 proliferation of downregulating Berlin1 expression

作　　者：张小薄[1] 高峰[1] 谭晓冬[1] 周磊[1] 王怀涛[1] 石刚[2] 

机构地区：[1]中国医科大学附属盛京医院外科,沈阳,110004 [2]辽宁省肿瘤医院大肠外科

出　　处：《中国综合临床》2015年第8期-,共5页Clinical Medicine of China

基　　金：国家自然科学基金资助项目

摘　　要：目的 观察下调自噬性相关蛋白(Beclin1)表达对抗凋亡基因(Bcl-2)表达的影响及胰腺癌细胞增殖变化,并探讨其机制.方法 构建靶向干扰质粒siBeclin1转染PANC-1细胞,细胞按照是否转染分为未转染组(Beclin1组)、空白对照组(siNegative control组)及转染组(siBeclin1组).以Western blot法及qRT-PCR法检测转染效率及Beclin1下调后Bcl-2变化;采用MTT检测siBeclin1转染后PANC-1细胞增殖.结果 Western Blot显示转染后Beclin1在siBeclin1组表达量明显低于siNegative Control组及Beclin1组(27.823±1.432、8.635±1.481、26.904±1.098,F=8.176,P＜0.01),qRT-PCR显示,与siNegativecontrol组及Beclin1组对比,转染siBeclin1后,PANC-1细胞Beclin1 mRNA表达量明显降低(0.421±0.157、0.194±0.104、0.399±0.123,F=5.239,P＜0.01),沉默效率为60％～70％;Bcl-2蛋白及mRNA在siBeclin1细胞的表达量明显高于Beclin1细胞(26.912±1.927、8.004±1.534,t=7.329,P＜0.01;0.582±0.297、0.217±0.186,t=6.835,P＜0.01);MTT显示,siBeclin1干扰后PANC-1细胞增殖能力增强,在72h分别为(61.54±6.81)％和(46.78±7.28)％,在96 h分别为(76.39±7.26)％和(54.27±8.17)％,差异均有统计学意义(t=3.674,P＜0.05;t=10.185,P＜0.01).结论 Beclin1表达下调后胰腺癌细胞增殖恶性行为增强,Beclin1可能通过下调Bcl-2表达抑制胰腺癌细胞的增殖,可作为胰腺癌治疗新的靶向候选基因.Objective To observe the influences of Bcl-2 expression and pancreatic cancer cell proliferation after downregulating Beclin1 expression and discuss the mechanism.Methods Target interfering plasmid siBeclin1 was constructed and transfected to PANC-1 cell,and cells were divided into non transfected group(Beclin1 group),blank control group(control siNegative group) and transfection group(siBeclin1 group).Western blot and qRT-PCR were used to detect the transfection efficiency and Bcl-2 expression after transfection.Proliferation was detected by MTT.Results Western blot and qRT-PCR results showed that expression of Beclin1 in siBeclin1 group was lower than Beclin1 and siNegative control groups (27.823± 1.432,8.635±1.481,26.904±1.098;F=8.176,P＜0.01),qRT-PCR showed that the Beclin1 mRNA level of PANC-1 cells in siBeclin1 group was lower than Belcin1 and siNegative groups (0.421 ±0.157,0.194±0.104,0.399 ±0.123;F=5.239,P＜0.01),and the silencing rate was about 60％-70％.Bcl-2 protein expression and levels of mRNA in siBeclin1 group were significantly higher than that of Beclin1 group(26.912±1.927,8.004±1.534,t =7.329,P＜0.01;0.582±0.297,0.217±0.186,t =6.835,P＜0.01);MTT results showed that the proliferation of PANC-1 cells were strengthened,(61.54±6.81)％ and (46.78±7.28)％ at 72 h,(76.39±7.26)％ and (54.27±8.17)％ at 96 h,and the difference was significant(t=3.674,P＜0.05;t =10.185,P ＜0.01).Conclusion Proliferation of pancreatic cancer cells are strengthened after Beclin1 expression downregulating,so Beclin1 may inhibit the proliferation of pancreatic cancer cells by Bcl-2 downregulating.Beclin1 can be a target candidate gene of pancreatic cancer therapy.

关 键 词：自噬相关性蛋白 胰腺癌 增殖 抗凋亡基因 

分 类 号：R5[医药卫生—内科学]