POLD1基因反义RNA对人正常肝细胞和肝癌细胞增殖的影响

Effect of POLD1 gene antisense RNA on proliferation of human normal liver cell and liver cancer cell

目的分析POLD1基因的反义RNA对人肝癌细胞SMMC-7721和人正常肝细胞HL-7702增殖的影响,以探讨利用POLD1基因反义RNA干预治疗肝癌的可行性。方法构建人POLD1基因的反义RNA表达质粒;将实验分为阴性对照组(转染空质粒的肝癌细胞SMMC-7721和肝细胞HL-7702)、空白对照组(未转染质粒的肝癌细胞SMMC-7721和肝细胞HL-7702)和实验组(转染人POLD1基因反义RNA表达质粒的肝癌细胞SMMC-7721和肝细胞HL-7702);采用CCK-8法分析细胞增殖情况。结果在转染0 h、24 h、48 h、72 h后,3组间肝细胞HL-7702的450 nm处吸光度值(A450值)差异无统计学意义(P>0.05)。实验组肝癌细胞SMMC-7721在转染24 h、48 h、72 h后,A450值均低于空白对照组和阴性对照组(P<0.05)。结论 POLD1基因的反义RNA可在对正常肝细胞生长无明显影响下抑制肝癌细胞的增殖。

Objective To analyze the effect of POLD1 gene antisense RNA on the proliferation of human liver cancer cell SMMC-7721 and human normal liver cell HL-7702 so as to explore the feasibility of POLD1 gene antisense RNA for the treatment of human liver cancer. Methods Expression plasmid with human POLD1 gene antisense RNA was constructed.The experiment consisted of negative control group（ HL-7702 cells and SMMC-7721 cells transfected with empty plasmid ） , blank control group （ untransfected HL-7702 cells and SMMC-7721 cells） and experimental group（ HL-7702 cells and SMMC-7721 cells transfected with human POLD1 gene antisense RNA） . Cell counting kit-8（CCK-8） assay was used to analyze the cell growth conditions.Results There was no significant difference in the absorbance value（A450 nm） of liver HL-7702 cells 0,24,48 or 72 hours after transfection among three groups（P〉0.05）.Compared with blank control group and negative control group, experimental group obtained lower absorbance values （ A450 nm ） of liver cancer cells SMMC-7721 24,48 and 72 hours after transfection（P〈0.05）.Conclusion POLD1 gene antisense RNA can inhibit the proliferation of liver cancer cells while the growth of normal hepatocytes is not affected.