摘要
目的运用RNA干扰(RNAinteference,RNAi)技术阻断胰腺癌细胞株PANC1中OCT4基因表达,并研究该基因沉默后对细胞增殖与凋亡的影响。方法利用阳离子脂质体LipofectamineTM2000将化学合成的人Octamer binding factor4(OCT4)的小干扰RNA转染入胰腺癌细胞株PANC1中。RT-PCR法测定胰腺癌细胞内OCT4mRNA的表达。WesternBlot测定OCT4、PARP。CCK8法测定细胞生长曲线观察细胞增殖的抑制情况。流式细胞仪测定细胞凋亡率的变化。结果化学合成的人OCT4siRNA能有效地抑制PANC1细胞中OCT4的表达(P<0.05)。OCT4siRNA组中细胞增殖减慢,凋亡率较对照组明显增加(P<0.05)。干扰OCT4能够激活PARP的表达。结论体外实验初步证明OCT4基因在胰腺癌细胞株增殖分化与凋亡方面扮演重要角色。通过沉默其表达可抑制胰腺癌细胞株PANC1增殖并诱导其凋亡。
Objective To observe the effects of small interfering RNA（siRNA） -mediated silencing of octamer binding factor 4（OCT4） gene on the proliferation and apoptosis of pancreatic carcinoma cell line PANC1 in vitro. Methods Chemically synthesized siRNA against human OCT4 was transfected into PANC1 cells via LipofectamineTM2000. The expression of OCT4 mRNA in the cells was detected using RT-PCR,and the protein expressions of OCT4 and PARP were assayed using Western blotting. The changes in the cell proliferation were evaluated using CCK8 method. Flow cytometry was used to detect the apoptosis of the transfected cells. Results siRNA transfection significantly suppressed the expression of OCT4 gene,which activated the expression of PARP in PANC1 cells（P0.05） . CCK8 method demonstrated a significant inhibition of cell pro1iferation by OCt4 siRNA transfection,which also resulted in significantly increased apoptotic rate of the cells（P0.05） . Conclusion siRNA-mediated OCT4 gene silencing can inhibit the proliferation and induce apoptosis of pancreatic carcinoma PANC1 cells,and this study provides a basis for further functional study of OCT4 gene.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2011年第5期860-863,共4页
Journal of Southern Medical University
基金
广州市科技局项目(2008Z3-E011)
