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SOX4基因抑制肝癌细胞增殖与侵袭能力
张岩,马丽辉,邓黎黎,张壮苗
0
(三亚市人民医院血液肿瘤内科)
摘要:
【摘要】目的 探讨小干扰RNA技术(siRNA)沉默SOX4基因对肝癌细胞增殖、侵袭能力的影响。方法 构建siRNA转染人肝癌细胞株SMMC7721,按转染质粒区别分为siRNASOX4组、siRNANC组,并设立空白对照组。实时荧光定量聚合酶联反应(RTPCR)检测各组细胞SOX4 mRNA,蛋白印迹法(WB)测定SOX4蛋白水平,噻唑蓝法(MTT)测定转染不同时间肝癌细胞增殖能力,流式细胞术测定肝癌细胞凋亡率,Transwell小室实验测定肝癌细胞侵袭能力。结果 转染后siRNASOX4组SOX4 mRNA相对表达量低于空白对照组和siRNANC组(P<005);转染后siRNASOX4组SOX4蛋白表达量为低于空白对照组和siRNANC组(P<005);转染不同时间siRNASOX4组细胞增殖活性均低于空白对照组和siRNANC组(P<005);转染后siRNASOX4组细胞凋亡率高于空白对照组和siRNANC组(P<005);转染后siRNASOX4组穿膜细胞比例低于空白对照组和siRNANC组(P<005)。结论 siRNA技术沉默SOX4基因可降低肝癌细胞增殖及侵袭能力,促进癌细胞凋亡。
关键词:  肝癌细胞  SOX基因  增殖  侵袭  凋亡  小干扰RNA技术
DOI:
基金项目:海南省三亚市科技局创新资助项目(2011YD88)
Experimental study on siRNA inhibiting proliferation and invasion ability of hepatoma cells by silencing SOX4 gene
ZHANG Yan,MA Lihui,DENG Lili,ZHANG Zhuangmiao
(Department of Hematology, Sanya People's Hospital)
Abstract:
【Abstract】Objective To explore effects of small interfering RNA (siRNA) silencing SOX4 gene on proliferation and invasion ability of hepatoma cells. Methods The siRNA was constructed to transfect into human hepatoma cell line SMMC7721. According to different transfection plasmid, they were divided into siRNASOX4 group and siRNANC group. And blank control group was set up. Realtime quantitative polymerase chain reaction (RTPCR) was performed to detect SOX4 mRNA in each group. Western blotting (WB) was performed to detect the expression of SOX4 protein. The proliferation ability of hepatoma cells with different transfection time was detected by methyl thiazolyl tetrazolium (MTT) assay. The apoptosis rate of hepatoma cells was detected by flow cytometry. The invasion ability of hepatoma cells was detected by Transwell chamber assay. Results After transfection, relative expression quantity of SOX4 mRNA in siRNASOX4 group was lower than that in blank control group and siRNANC group [(2045±0056) vs (8961±0452), (8321±0512)] (P<005). After transfection, expression quantity of SOX4 protein in SOX4 group was lower than that of blank control group and siRNANC group [(0263±0024) vs (1006±0086), (0999±0079)] (P<005). At different transfection time, cell proliferation activity in siRNASOX4 group was lower than that in blank control group and siRNANC group (P<005). After transfection, apoptosis rate of siRNASOX4 group was higher than that of blank control group and siRNANC group [(2612±354)% vs (1031±121)%, (997±133)%] (P<005). After transfection, ratio of transmembrane cells in siRNASOX4 group was lower than that in blank control group and siRNANC group [(2563±673)% vs (6012±1054)%, (5997±1161)%] (P<005). Conclusion Silencing SOX4 gene by siRNA can reduce proliferation and invasion ability of hepatoma cells, and promote apoptosis of cancer cells.
Key words:  Hepatoma cell  SOX gene  Proliferation  Invasion  Apoptosis  Small interfering RNA technology

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