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作 者:傅春江 毛廷森 曾泉[2] 陈琳[2] 周军年[2] 贾雅丽[2] 岳文[2]
机构地区:[1]房山区中医院,北京102400 [2]军事医学科学院野战输血研究所全军干细胞与再生医学重点研究室,北京100850
出 处:《生物技术通讯》2015年第5期600-604,共5页Letters in Biotechnology
基 金:国家重点基础研究计划(2011CB64804)
摘 要:目的:在肝癌细胞系SK-HEP-1及MHCC-97H中过表达mi R-155,研究其对肝癌细胞增殖能力的影响。方法:将pc DNA3.0-mi R-155表达载体瞬时转染SK-HEP-1及MHCC-97H肝癌细胞系,通过实时定量PCR技术检测mi R-155在转录水平的表达;采用CCK8法及克隆形成实验检测mi R-155过表达后对SK-HEP-1及MHCC-97H肝癌细胞系增殖的影响;采用Transwell实验探讨mi R-155过表达后对肿瘤细胞浸润能力的影响。结果:实时定量PCR检测结果显示,转染SK-HEP-1及MHCC-97H细胞后72 h,成熟mi R-155表达分别上调约424±48.5倍及63.69±7.8倍,表明mi R-155在肿瘤细胞内得到高表达;CCK8法及克隆形成实验结果显示,mi R-155能够明显促进肝癌细胞增殖(P<0.01);Transwell实验证明mi R-155明显促进MHCC-97H肿瘤细胞的浸润能力。结论:mi R-155在肝癌细胞中的高表达能够明显促进肝癌细胞增殖与浸润,提示其有可能在肝癌的发生发展中发挥重要作用。Objective: To study whether miRNA-155 influence hepatocellular carcinoma cell biology, we over ex- pressed miRNA-155 in SK-HEP-1 and MHCC-97H cells and checked the proliferation and invasion activity of cells. Methods: SK-HEP-1 and MHCC-97H cells with ectopic miRNA-155 expression were constructed and iden- tified by real-time PCR. CCK8 analysis and colony formation assay were used to detect cancer cell proliferation. The invasion activity of cancer cells were detected by Matrigel-transwell assay. Results: Our results suggested that ectopic expression of miRNA-155 in SK-HEP-I and MHCC-97H cell could be highly expressed after trans- fection, and the expression level was accordingly upregulated 424±48.5 and 63.69±7.8 folds respectively. CCK8 as- say and colony-forming unit assay also showed that miR-155 could obviously promote hepatoeellular carcinoma cell proliferation(P〈0.01). Matrigel-transwell assay result suggested that miR-155 could enhance the cell invasion activity significantly. Conclusion: Taken together, our results reveal that miRNA-155 is involved in hepatoeellular carcinoma cell progression and may be a potential therapeutic target of cancer cells.
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