CAJ | 학술논문

目的：探讨针对FOXO3a的小干扰RNA（siRNA）对HepG2 FOXO3a的表达、细胞增殖的抑制及细胞凋亡的作用。方法利用RNA干扰（RNAi）效应设计针对FOXO3a的不同siRNA，构建表达载体质粒并转染HepG2细胞系。RT-PCR和Western Blot分别检测FOXO3a mRNA及蛋白的表达，细胞生长实验和流式细胞观察转染前后细胞增殖及细胞凋亡的变化。另设一无意义RNA载体质粒为阴性对照。结果 P2、P3实验组，细胞核绿色荧光明显减淡，有细胞核的排出。特异性siRNA对FOXO3a mRNA及蛋白的表达具有抑制作用（P<0.05），细胞凋亡减少，增殖明显增加（P<0.05）。结论抑制 FOXO3a 基因的表达能够减少HepG2细胞的凋亡，并增加细胞的增殖。FOXO3a有望成为治疗肝癌的有效的靶基因。
목적：탐토침대FOXO3a적소간우RNA（siRNA）대HepG2 FOXO3a적표체、세포증식적억제급세포조망적작용。방법이용RNA간우（RNAi）효응설계침대FOXO3a적불동siRNA，구건표체재체질립병전염HepG2세포계。RT-PCR화Western Blot분별검측FOXO3a mRNA급단백적표체，세포생장실험화류식세포관찰전염전후세포증식급세포조망적변화。령설일무의의RNA재체질립위음성대조。결과 P2、P3실험조，세포핵록색형광명현감담，유세포핵적배출。특이성siRNA대FOXO3a mRNA급단백적표체구유억제작용（P<0.05），세포조망감소，증식명현증가（P<0.05）。결론억제 FOXO3a 기인적표체능구감소HepG2세포적조망，병증가세포적증식。FOXO3a유망성위치료간암적유효적파기인。
Objective To explore the effects of FOXO3a siRNAs on FOXO3a expression,cell cirele and apoptosis of HepG2 cells.Methods Three FOXO3a specific siRNAs were selected with the suppressing effect. RNAi expression vectors were constructed and transfected into HepG2 cells.RT-PCR and Western Blot were used to detect the expression of FOXO3a.The proliferation of the cells was measured by MTT test.The apoptosis of HepG2 cells was measured by flow cytometry.A nonsense small RNA was set as negative control. Results There were the export of FoxO3a to the cytoplasm from nucleus in P2,P3 group. When compared with the control groups,the FOXO3a-specific siRNAs inhibited the expression of FOXO3a in the protein and mRNA level, and suppressed the proliferation and induced the apoptosis of the HepG2 cells (P<0.05).Conclusion Knockdown expression of FOXO3a can decrease apoptosis and increase proliferation of HepG2 tumor cell,and FOXO3a gene can be a powerful target gene in treating liver cancer.