摘要
目的探讨微小RNA-9(miR-9)调节同源异形盒1(HMBOX1)基因对胃癌细胞生物学功能影响的作用。方法将对数生长期的胃癌细胞株BGC-823细胞分为空白组、对照组与miR-9组,对照组与miR-9组分别转染50 nmol/L的miR-9 NC、miR-9 mimics,空白组不进行转染(加入等体积的磷酸盐缓冲液)。采用实时荧光定量聚合酶链反应法检测miR-9 mRNA的表达,噻唑蓝法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,蛋白质印迹法检测HMBOX1蛋白表达情况,上述实验均重复3次,计算其平均值。结果细胞转染后24 h与48 h,miR-9组的miR-9 mRNA相对表达量高于空白组和对照组[(46. 45±11. 82)比(1. 00±0. 12)、(1. 03±0. 13),(55. 45±10. 21)比(1. 02±0. 22)、(1. 11±0. 32)],细胞凋亡指数高于空白组和对照组[(42. 6±1. 5)%比(11. 6±2. 1)%、(13. 2±1. 2)%,(48. 3±1. 5)%比(17. 4±2. 4)%、(15. 3±1. 8)%](均P <0. 05)。细胞转染后24 h与48 h,miR-9组的细胞增殖指数低于空白组和对照组[(0. 42±0. 13)比(1. 11±0. 78)、(1. 07±0. 51),(0. 53±0. 13)比(1. 17±0. 22)、(1. 12±0. 12)],HMBOX1蛋白相对表达水平低于空白组和对照组,差异均有统计学意义(均P <0. 05)。结论过表达miR-9能靶向抑制HMBOX1的表达,从而抑制胃癌细胞增殖,促进细胞凋亡,发挥抑癌作用。
Objective To investigate the effect of microRNA-9( miR-9) on the biological function of gastric cancer cells by regulating HMBOX1. Methods Gastric cancer cell line BGC-823 was divided into blank group,control group and miR-9 group. The control group and miR-9 group were transfected with 50 nmol/L miR-9 NC and miR-9 mimics,respectively;the blank group was treated with equal volume of phosphate buffer. Expression of miR-9 mRNA was detected by real-time fluorescence quantitative polymerase chain reaction. Cell proliferation was detected by methylthiazoletetrazolium assay. Cell apoptosis was detected by flow cytometry. HMBOX1 expression was detected by western blotting. Above experiments were repeated three times to obtain the average value.Results At 24 h and 48 h after transfection,expression of miR-9 and cell apoptosis index in the miR-9 group were significantly higher than those in the blank group and control group[( 46. 45 ± 11. 82) vs( 1. 00 ± 0. 12),( 1. 03 ± 0. 13);( 55. 45 ± 10. 21) vs( 1. 02 ± 0. 22),( 1. 11 ± 0. 32);( 42. 6 ± 1. 5) % vs( 11. 6 ± 2. 1) %,( 13. 2 ± 1. 2) %;( 48. 3 ± 1. 5) % vs( 17. 4 ± 2. 4) %,( 15. 3 ± 1. 8) % ]( all P < 0. 05);cell proliferation index in the miR-9 group was significantly lower than that in the blank group and control group [( 0. 42 ± 0. 13) vs( 1. 11 ± 0. 78),( 1. 07 ± 0. 51);( 0. 53 ± 0. 13) vs( 1. 17 ± 0. 22),( 1. 12 ± 0. 12) ]( all P < 0. 05);expression of HMBOX1 in the miR-9 group was significantly lower than that in the blank group and control group( all P <0. 05). Conclusion Over-expression of miR-9 can inhibit the expression of HMBOX1,thereby inhibiting the proliferation of gastric cancer cells and promote apoptosis.
作者
朱隽
余华良
高山
丁祥武
Zhu Jun;Yu Hualiang;Gao Shan;Ding Xiangwu(Department of Digestive Medicine,North District of Xiangyang Central Hospital,Hubei Province,Xiangyang 441000,China;Department of Digestive Medicine,Ping An Hao Yi Xiangyang Comprehensive Clinic,Xiangyang 441000,China;Department of Digestive Medicine,Xiangyang Central Hospital,the Affiliated Hospital of Hubei University of Arts and Science,Hubei Province,Xiangyang 441021,China)
出处
《中国医药》
2020年第1期83-86,共4页
China Medicine
基金
湖北省卫生和计划生育委员会科研立项项目(WJ2015MB184)~~
