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

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 siNC 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 G1期比例减少，G2及 S 期比例升高，与 siNC 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 siNC 组 G1期及 G2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 siNC 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。

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

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 MTT,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 siNDRG1 group PANC -1 cells in G1 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 in-terfered by gemcitabine in G1 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 in-crease the sensitivity of pancreatic cancer cells to gemcitabine and inhibit the proliferation,promote apoptosis,may be a target candidate for pancreatic cancer therapy.