CAJ | 학술논문

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.
목적 관찰소분자간우RNA(siRNA)침묵성별결정기인상관HMG합기인-6(SOX-6)대체외효질류세포증식능력적영향.방법 인효질류모세포세포류LN229세포주체외배양후,분위3조,실험조、대조조분별이SOX-6 siRNA、Control siRNA-A전염후배양,공백조상규배양,채용면역조직화학염색화면역형광법감정SOX-6 siRNA전염성공솔,채용세포계수시제합(CCK-8)법검측세포증식개변.결과 실험조화대조조SiRNA전염솔＞80％;역전록-취합매련반응(RT-PCR)결과현시,여대조조급공백조비교,실험조SOX-6표체현저강저(0.427±0.032비0.612±0.055비0.609±0.049),CCK-8실험검측결과현시,여대조조화공백조비교,실험조흡광도(A450)치재24、48、72 h균명현강저(P＜0.01).결론 채용siRNA침묵SOX-6기인가조공효질류세포적체외증식.
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.