NDRG1下调对吉西他滨干预人胰腺癌细胞PANC-1增殖及凋亡的影响  

NDRG1 down- regulating influence proliferation and apoptosis of PANC- 1 cells interfered with gemcitabine

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

机构地区：[1]中国医科大学附属盛京医院普通外科,辽宁沈阳110004 [2]辽宁省肿瘤医院大肠外科,辽宁沈阳110042

出　　处：《现代肿瘤医学》2016年第1期26-30,共5页Journal of Modern Oncology

基　　金：辽宁省自然科学基金资助项目(编号:2014021006)

摘　　要：目的:观察siNDRG1干扰对吉西他滨干预的胰腺癌细胞增殖、细胞周期及凋亡的影响并探讨其机制。方法:以Western blot法对PANC-1、BXPC-3、CAPAN-2、SW1990细胞中NDRG1的表达进行检测;构建靶向NDRG1的干扰质粒siNDRG1,转染PANC-1细胞,以Western blot法及(qRT)-PCR法检测NDRG1干扰效率;对siRNA干扰细胞给予吉西他滨干预,采用MTT法检测细胞增殖,PI法检测细胞周期,流式细胞术检测细胞调亡。结果:Western blot显示,NDRG1在四组细胞株中均有表达,低分化细胞(PANC-1)中表达量较中分化(BXPC-3)及高分化细胞株(CAPAN-2、SW1990)中高(P<0.05);MTT显示,吉西他滨干预后PANC-1细胞在siNDRG1转染组细胞生长抑制率高于si NC组,差异有显著性(P<0.01)。PI法显示,转染siNDRG1的PANC-1细胞停留在G_1期比例减少,G_2及S期比例升高,与si NC组对比,差异具有显著性(t=4.21,P<0.05;t=3.54,P<0.05;t=3.29,P<0.05)。经吉西他滨处理后,siNDRG1较si NC组G_1期及G_2期细胞比例明显减少,S期细胞比例升高,差异具有显著性(t=14.65,P<0.01;t=13.28,P<0.01;t=15.17,P<0.01)。流式细胞仪检测经吉西他滨处理后siNDRG1组凋亡率高于si NC组,差异有显著性(t=22.42,P<0.001)。结论:对胰腺癌细胞NDRG1表达进行下调可以增强吉西他滨化疗敏感性,抑制细胞增殖,促进凋亡,可作为胰腺癌治疗新的靶向候选基因。Objective：To observe the effect of NDRG1 down -regulating on proliferation, cell cycle and apoptosis of pancreatic cells interfered by gemcitabine and discuss the mechanism. Methods ： Western blot was used to detect the NDRG1 expressions in PANC - 1, BXPC - 3, CAPAN - 2, SW1990 cells. Target siNDRG1 was constructed and transfected into PANC - 1 cells to downregulate the expression of NDRG1. Interfering effect was detected by Western blot and （qRT） -PCR. siRNA interfering cells was dealt with gemcitabine. Proliferation, cell cycle and apoptosis were detected by M3T, PI method and flow cytometry. Results：Western blot showed that NDRG1 was expressed in PANC - 1 ,BXPC -3 ,CAPAN -2 and SW1990 cells ,the expression was higher in low differentiation cell（ PANC -1 ） than in middle differentiation （ BXPC - 3 ） and high differentiation （ CAPAN - 2, SW1990 ） cells （ P 〈 0.05 ）. MMT showed that the inhibition rate of PANC - 1 cell interfered by gemcitabine in siNDRG1 group was higher than in siNC group （ P 〈 0.01 ）. PI method showed that in siNDRGI group PANC - 1 cells in Gt phase was decreased,in G2 and S phases were increased （ t = 4. 21 ,P 〈 0. 05, t = 3.54, P 〈 0. 05, t = 3.29, P 〈 0.05 ）. In siNDRG1 group, PANC - 1 cell interfered by gemcitabine in Gl and G2 phases were decreased and in S phase was increased（t = 14. 65,P〈0. 01, t = 13.28,P 〈 0. 01, t = 15. 17, P 〈 0.01 ）. Flow cytometry showed that the apoptosis rate in siNDRG1 group deal with gemcitabine were higher than in siNC group（ t = 22. 42 ,P 〈 0. 001 ）. Conclusion： NDRG1 down - regulating can increase the sensitivity of pancreatic cancer ceils to gemcitabine and inhibit the proliferation, promote apoptosis, may be a target candidate for pancreatic cancer therapy.

关 键 词：NDRG1 胰腺癌 吉西他滨 调亡 细胞周期 

分 类 号：R735.9[医药卫生—肿瘤]