CAJ | 학술논문

目的：应用RNA干扰技术抑制人肝癌细胞SMMC-7721中POLD1基因的表达，探索该技术干预肝癌的可行性。方法制备4个针对人POLD1基因的shRNA表达质粒，转染SMMC-7721细胞48 h后，荧光定量PCR法检测POLD1基因的表达，CCK-8检测细胞生长情况。结果测序鉴定证实4个shRNA表达质粒构建成功，将其转染SMMC-7721细胞后，NEO-POLD1-1和NEO-POLD1-4表达质粒抑制效果明显，使SMMC-7721细胞的增殖受到抑制，并且使POLD1基因表达在mRNA水平抑制，NEO-POLD1-1相对表达量为（0．1425±0．0205）， NEO-POLD1-4相对表达量为（0．209±0．009）。结论针对POLD1基因设计的shRNA在体外有效地抑制了POLD1基因mRNA的表达和肝癌细胞的增殖，且实现RNA干扰具有序列选择性。
목적：응용RNA간우기술억제인간암세포SMMC-7721중POLD1기인적표체，탐색해기술간예간암적가행성。방법제비4개침대인POLD1기인적shRNA표체질립，전염SMMC-7721세포48 h후，형광정량PCR법검측POLD1기인적표체，CCK-8검측세포생장정황。결과측서감정증실4개shRNA표체질립구건성공，장기전염SMMC-7721세포후，NEO-POLD1-1화NEO-POLD1-4표체질립억제효과명현，사SMMC-7721세포적증식수도억제，병차사POLD1기인표체재mRNA수평억제，NEO-POLD1-1상대표체량위（0．1425±0．0205）， NEO-POLD1-4상대표체량위（0．209±0．009）。결론침대POLD1기인설계적shRNA재체외유효지억제료POLD1기인mRNA적표체화간암세포적증식，차실현RNA간우구유서렬선택성。
Objective To investigate the feasibility of RNA interfering ( RNAi ) on liver cancer by using this technique to inhibit the expression of POLD 1 gene in human liver cancer cells SMMC-7721.Methods Four expression plasmids with shRNAs targeting POLD 1 gene were constructed ,and then the plasmids were transfected to SMMC-7721 for 48 hours.The expression of POLD1 gene was detected by real-time RT-PCR.CCK-8 assay was used to analyze the change of cell proliferation .Results The four expression plasmids were constructed successfully , confirmed by sequencing .After the plasmids were transfected to SMMC-7721 , the cell proliferation was significantly inhibited in NEO-POLD1-1 and NEO-POLD1-4.The mRNA expression of POLD1 gene in NEO-POLD1-1 and NEO-POLD1-4 were significantly inhibited as (0.1425 ±0.0205) and (0.209 ±0.009).Conclusion shRNAs targeting the POLD1 gene can inhibit the mRNA expression of POLD1 gene and liver cancer cell proliferation in vitro ,and the RNA interfering is sequencing alternative .