POLD1基因反义RNA对人肝癌细胞SMMC-7721的抑制作用  

Inhibitory effects of POLD1 gene antisense RNA on human hepatoma cells SMMC-7721

作　　者：吴琼[1] 安娜[1] 廖柳凤[1] 黄文涛[2] WU Qiong;AN Na;LIAO Liu-feng;HUANG Wen-tao(Department of Clinical Pharmacy,Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530000,Guangxi Zhuang Autonomous Region,China;Department of Pharmacy,Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学附属肿瘤医院临床药学科,广西南宁530000 [2]广西医科大学药学系,广西南宁530021

出　　处：《中国临床药理学杂志》2021年第16期2148-2151,共4页The Chinese Journal of Clinical Pharmacology

基　　金：广西医科大学教育厅2016年中青年教师基础能力提升基金资助项目(KY2016LX037);广西医科大学青年科学基金资助项目(GXMUYSF201631)。

摘　　要：目的研究DNA聚合酶δ催化亚基基因1(POLD1)反义RNA对人肝癌细胞SMMC-7721的抑制作用及相应机制。方法将SMMC-7721细胞分为3组:实验组、阴性对照组、空白组。实验组及阴性对照组分别将POLD1基因反义RNA表达质粒及空质粒分别转染至SMMC-7721细胞,未经处理的SMMC-7721细胞为空白组。用聚合酶链反应法检测各组细胞POLD1基因表达情况;用细胞计数-8(CCK-8)法检测细胞增殖情况;用流式细胞术分析细胞凋亡率;用蛋白质印迹(Western blot)法检测p53、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)等凋亡相关蛋白表达量。结果实验组、阴性对照组和空白组肝癌细胞POLD1基因相对表达量分别为0.18±0.03,1.03±0.18和1.00±0.00;转染72 h增殖情况(OD_(450)值)分别为0.57±0.06,0.73±0.09和0.77±0.12;凋亡率分别为(23.43±4.12)%,(9.12±1.03)%,(1.24±0.14)%,实验组的上述指标与阴性对照组和空白组比较,差异均有统计学意义(均P<0.05)。结论POLD1基因反义RNA可显著抑制人肝癌细胞SMMC-7721的增殖,且能诱导人肝癌细胞SMMC-7721凋亡。Objective To study the inhibitory effects and corresponding mechanism of DNA polymerase δ catalytic subunit gene 1(POLD1) antisense RNA on human hepatoma cells SMMC-7721. Methods SMMC-7721 cells were divided into experimental group, negative control group and blank group. Experimental group and negative control group were transfected the expression plasmid and empty plasmid of POLD1 gene antisense RNA into SMMC-7721 cells. The untreated SMMC-7721 cells were selected as blank group. The expression of POLD1 gene in each group was detected by polymerase chain reaction. The cells proliferation was detected by cell counting kit-8(CCK-8). The apoptosis rate was analyzed by flow cytometry. The expression levels of apoptosis-related proteins [P53, B-cell lymphoma-2(Bcl-2), Bcl-2 associated X protein(Bax)]were detected by Western blot. Results The relative expression levels of POLD1 gene in experimental group, negative control group and blank group were 0.18±0.03, 1.03±0.18 and 1.00±0.00, the 72 h proliferation rate(OD450) were 0.57±0.06, 0.73±0.09 and 0.77±0.12, the apoptosis rates were ( 23. 43 ± 4. 12) %,( 9. 12 ± 1. 03) % and( 1. 24 ± 0. 14) %. The differences in the above indexes between experimental group and negative control group,blank group were statistically significant( all P < 0. 05). Conclusion POLD1 gene antisense RNA can significantly inhibit the proliferation of human hepatoma cells SMMC-7721,and induce their apoptosis.

关 键 词：肝癌细胞 DNA聚合酶δ催化亚基基因1 反义RNA 细胞抑制 

分 类 号：R979.1[医药卫生—药品]