CAJ | 학술논문

目的：获得敲低效果较好的CCCTC结合因子（CTCF)的RNA干扰腺病毒载体，以便于研究其在肿瘤发生发展中的作用。方法：从已发表文献中获得CTCF敲低靶序列，合成2对含有小发卡结构的寡核苷酸序列，将其进行退火磷酸化后，分别克隆到腺病毒包装载体上；将重组质粒转染人胚肾293A细胞，收获腺病毒；将收获的腺病毒分别感染人胚肾HEK293细胞和靶细胞人肺腺癌细胞A549，通过RT-PCR和Western印迹鉴定相关基因的表达变化。结果与结论：RT-PCR和Western印迹鉴定显示构建的表达CTCF短发夹RNA（shRNA）的腺病毒载体能够有效抑制CTCF转录和蛋白水平，为后续CTCF的生物学功能和机制研究奠定了基础。
목적：획득고저효과교호적CCCTC결합인자（CTCF)적RNA간우선병독재체，이편우연구기재종류발생발전중적작용。방법：종이발표문헌중획득CTCF고저파서렬，합성2대함유소발잡결구적과핵감산서렬，장기진행퇴화린산화후，분별극륭도선병독포장재체상；장중조질립전염인배신293A세포，수획선병독；장수획적선병독분별감염인배신HEK293세포화파세포인폐선암세포A549，통과RT-PCR화Western인적감정상관기인적표체변화。결과여결론：RT-PCR화Western인적감정현시구건적표체CTCF단발협RNA（shRNA）적선병독재체능구유효억제CTCF전록화단백수평，위후속CTCF적생물학공능화궤제연구전정료기출。
Objective: To obtain high effective small interfering RNA(siRNA) interference vector of CCCTC bind-ing factor(CTCF) gene for studying the biological function of CTCF in the development and progression of tumor. Methods: CTCF siRNA was designed and constructed based on published papers and the synthesized sequences were annealed to form double-strand oligonucleotide and cloned into interference vector. Then the interference vec-tors were transfected into 293A cells. The knockdown effect was detected by RT-PCR and Western blotting. Re-sults & Conclusion: The adenoviruses containing short hairpin RNA(shRNA) targeting the CTCF gene have been successfully constructed, which laid the foundation for further study of CTCF function.