CAJ | 학술논문

目的：构建Peroxiredoxin2（PRDX2）基因RNA干扰（RNAinterference，RNAi）慢病毒表达载体，探讨PRDX2基因干扰后对结直肠癌SW480细胞增殖的影响。方法设计、合成靶向PRDX2的RNA干扰的序列，构建pGC‐EGFP‐shPRDX2慢病毒载体并进行鉴定，同时应用qRT‐PCR和Westernblot方法观察转染的SW480结肠癌细胞PRDX2mRNA和蛋白表达的抑制效果，并通过MTT、平板克隆形成实验检测细胞增殖变化。结果成功构建PRDX2基因慢病毒载体并经测序证实；pGC‐EGFP‐shPRDX2可有效抑制结直肠癌SW480细胞PRDX2的表达，感染慢病毒的SW480细胞中PRDX2mRNA和蛋白表达水平明显降低（P＜0．05）；SW480细胞经PRDX2RNA干扰后其生长和增殖能力显著降低（P＜0．05）。结论PRDX2基因RNAi慢病毒表达载体在SW480细胞中表达稳定可靠，PRDX2基因干扰后有效抑制了结直肠癌SW480细胞的增殖和生长，为进一步探讨PRDX2在结直肠癌发生、发展及转移中的作用奠定基础。
목적：구건Peroxiredoxin2（PRDX2）기인RNA간우（RNAinterference，RNAi）만병독표체재체，탐토PRDX2기인간우후대결직장암SW480세포증식적영향。방법설계、합성파향PRDX2적RNA간우적서렬，구건pGC‐EGFP‐shPRDX2만병독재체병진행감정，동시응용qRT‐PCR화Westernblot방법관찰전염적SW480결장암세포PRDX2mRNA화단백표체적억제효과，병통과MTT、평판극륭형성실험검측세포증식변화。결과성공구건PRDX2기인만병독재체병경측서증실；pGC‐EGFP‐shPRDX2가유효억제결직장암SW480세포PRDX2적표체，감염만병독적SW480세포중PRDX2mRNA화단백표체수평명현강저（P＜0．05）；SW480세포경PRDX2RNA간우후기생장화증식능력현저강저（P＜0．05）。결론PRDX2기인RNAi만병독표체재체재SW480세포중표체은정가고，PRDX2기인간우후유효억제료결직장암SW480세포적증식화생장，위진일보탐토PRDX2재결직장암발생、발전급전이중적작용전정기출。
Objective To construct a lentiviral expression vector of peroxiredoxin2(PRDX2) RNA interference (RNAi) and to investigate the effect of siRNA of PRDX2 genes on the proliferation of human colonrectal cancer SW480 cell .Methods RNAi tar‐get sequences were designed and synthesized towards the PRDX2 gene sequences .The lentiviral vector pGC‐EGFP‐shPRDX2 was constructed and identified .The vector was transformed into SW480 cells ,and the transfection efficiency was evaluated by fluores‐cence microscopy .The expression of PRDX2 was detected with Quantitative real‐time PCR (qRT‐PCR) and Western blot in the transfected cells .Cell growth and colony forming ability were detected with MTT and plate cloning technique .Results PRDX2 gene lentiviral vector was successfully established and was proved by gene sequencing .The expression of PRDX2 in mRNA and pro‐tein was significantly reduced(P<0 .05) .The PRDX2 mRNA and protein expression in SW480 transfected with lentiviral were sig‐nificantly reduced (P< 0 .05) ,and the ability of growth and proliferation were significantly reduced(P< 0 .05) .Conclusion PRDX2 gene lentiviral vector could be a stable and reliable tool .The proliferation and growth of SW480 cells transfected by pGC‐EGFP‐shPRDX2 could be effectively suppressed ,which could facilitate further investigation of the roles of PRDX2 gene in the de‐velopment and progression of colorectal cancer .