SFRP5基因沉默促进人类胰腺癌细胞系PANC-1增殖与迁移

目的探讨慢病毒介导短发夹RNA(shRNA)沉默SFRP5基因对人类胰腺癌细胞系PANC-1细胞增殖、侵袭和迁移能力的影响。方法构建靶向SFRP5基因特异性shRNA慢病毒载体并转染人胰腺癌PANC-1细胞系,以空白质粒转染阴性对照组,未处理细胞做为空白对照组。用real-time PCR及Western blot检测转染前后SFRP5 RNA以及蛋白的表达;CCK-8实验检测细胞体外增殖能力;使用Transwell小室实验分析细胞侵袭能力;细胞划痕实验分析细胞迁移能力。结果成功建立稳定转染shRNA-SFRP5胰腺癌PANC-1细胞株。SFRP5病毒转染组与阴性对照组及空白对照组相比细胞的增殖能力明显增加(P0.01);SFRP5病毒转染组的细胞侵袭、迁移能力明显高于阴性对照组及空白对照组(P0.01)。结论 SFRP5慢病毒干扰载体能有效抑制SFRP5基因在人胰腺癌PANC-1细胞中的表达,进而促进细胞的增殖、侵袭和迁移。

SFRP5 gene silencing promotes proliferation and migration of human pancreatic cancer cell line PANC-1

Objective To investigate the effect of lentivirus-mediated shRNA silencing SFRP5 on proliferation,in-vasion and migration of pancreatic cancer cell line PANC-1. Methods A SFRP5-knockdown recombinant plasmid was constructed and successfully transfected it into pancreatic cancer cell line PANC-1,blank plasmid transfection was treated as negative control and untreated cells as blank control group. The expression of SFRP5 at RNA and protein level in cell were detected by real-time PCR and Western blot, CCK-8 assay was applied to examine the effect of SFRP5 silencing on the proliferation, the cell migration of pancreatic cancer cell line PANC-1 was ana-lyzed by Transwell migration assay and cell scratch test was used to examine the cell invasion in PANC-1 cell. Results Stable transfected shRNA-SFRP5 cell of pancreatic cancer line was established successfully.The prolifera-tion capacity of SFRP5 group was significantly higher as compared to the negative control and blank control group by CCK8 assay(P<0.01).Similarly, cell invasion and migration of SFRP5 group were significantly higher compared to the negative control and blank control group(P<0.01). Conclusions SFRP5 lentiviral interference vectors can effectively decrease SFRP5 gene expression in PANC-1 cell of pancreatic cancer, thereby promoting cell proliferation,invasion and migration.