沉默性别决定基因相关HMG盒基因-6对调控胶质瘤细胞增殖的研究

Silencing SRY-related high-mobility-group box-6 gene regulating proliferation of glioma cells in vitro

目的 观察小分子干扰RNA（siRNA）沉默性别决定基因相关HMG盒基因-6（SOX-6）对体外胶质瘤细胞增殖能力的影响.方法 人胶质瘤母细胞细胞瘤LN229细胞株体外培养后,分为3组,实验组、对照组分别以SOX-6 siRNA、Control siRNA-A转染后培养,空白组常规培养,采用免疫组织化学染色和免疫荧光法鉴定SOX-6 siRNA转染成功率,采用细胞计数试剂盒（CCK-8）法检测细胞增殖改变.结果 实验组和对照组SiRNA转染率＞80％;逆转录-聚合酶链反应（RT-PCR）结果显示,与对照组及空白组比较,实验组SOX-6表达显著降低（0.427±0.032比0.612±0.055比0.609±0.049）,CCK-8实验检测结果显示,与对照组和空白组比较,实验组吸光度（A450）值在24、48、72 h均明显降低（P＜0.01）.结论 采用siRNA沉默SOX-6基因可调控胶质瘤细胞的体外增殖.

Objective To investigate the effects of small interfering RNA （siRNA）-induced specific silence of SRY-related high-mobility-group box-6 （SOX-6） gene on the proliferation of glioblastoma cells in vitro.Methods The LN229 cells in good conditions were selected to be transfected by SOX-6 silenced by small interfering RNA with fluorescent marker,and observed under the fluorescent microscope to check the transfection efficiency.The cells were divided into 3 groups：transfection of control siRNA-A as control group; transfection of SOX-6 silenced by small interfering RNA as experimental group,and normal culture as blank control group.The expression of SOX-6 as well as its transcriptional activity was examined using reverse transcription-polymerase chain reaction （RT-PCR）.The proliferation of cells was analyzed by using counting kit-8 （CCK-8） assay.Results After transfection,RT-PCR results showed that SOX-6 gene expression in LN229 cells was inhibited specifically in experimental group as compared with control group and blank group （0.427 ± 0.032 vs.0.612 ± 0.055 vs.0.609 ± 0.049）.CCK-8 assay results showed that cell proliferation in experimental group was decreased significantly as compared with the control group and blank group at 24,48 and 72 h （P 〈 0.01）.Conclusion SOX-6 gene plays an important role in regulating the proliferation of LN229 cells in vitro.