摘要
目的 观察小干扰RNA （siRNA）沉默Kruppel样转录因子8（KLF8）基因对胃癌细胞MKN-28增殖及侵袭能力的影响.方法 构建针对KLF8的小干扰RNA（si-KLF8）,应用实时定量聚合酶链反应（Real-time PCR）和Westem blot技术检测转染后胃癌细胞中KLF8 mRNA和蛋白的表达,应用噻唑蓝（MTT）比色法检测转染24、48、72 h的细胞增殖能力的变化,应用Transwell小室实验检测转染48 h后胃癌细胞侵袭能力的变化.结果 si-KLF8和阴性对照siRNA （si-CTRL）成功转染至胃癌细胞;si-KLF8组KLF8 mRNA和蛋白的表达量明显低于阴性对照组和正常对照组（P＜0.05）;si-KLF8组增殖能力下降,24、48、72 h的吸光度值分别为0.125±0.003、0.193±0.005、0.223±0.005、0.267 ±0.003、0.315±0.006,穿膜细胞数为（42.0±3.6）个,均明显低于阴性对照组和正常对照组（P＜0.05）;阴性对照组和正常对照组比较,上述指标差异无统计学意义（P＞0.05）.结论 si-KLF8可抑制人胃癌细胞的增殖和侵袭能力,提示KLF8在胃癌的发生发展过程中发挥着重要的作用.
Objective To investigate the effects of small interference RNA （siRNA） targeting Kruppel like factor 8 （KLF8） gene （si-KLF8） on proliferation and invasion of gastric cancer cell MKN-28.Methods Synthesized siRNA targeting KLF8 was transfected into MKN-28,at the same time,gastric cancer cells transfected with negative siRNA were used as control.KLF8 mRNA and protein were detected by using real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting respectively at 24 h.The viability of cells was examined by using methyl thiazol tetrazolium （MTT） assay at 24,48 and 72 h.The cell invasion were assessed by Transwell assay.Results Si-KLF8 and negative control siRNA （si-CTRL） were transfected into gastric cancer cells successfully.KLF8 mRNA and protein in si-KLF8 group were lower than in negative control group and normal control group.The proliferations of cells were 0.125 ±0.003,0.193 ± 0.005,0.223 ±0.005,0.267 ± 0.003 and 0.315 ± 0.006 at 24,48 and 72 h,which were significantly lower than those in negative control group and normal control group.Transwell assay showed that the cell numbers migrated to Matrigel in si-KLF8 group （42.0 ± 3.6） were significantly lower than in negative control group and normal control group.However,no significant difference was found between negative control group and normal control group.Conclusion si-KLF8 can inhibit proliferation and invasion capability of human gastric cancer cells,which indicate KLF8 plays an important role in the generation and development of gastric cancer.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第2期316-318,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81172029、81172026、81402403）
国家重大科技专项（863）资助项目（2012AA022606）
上海交通大学医学院附属新华医院院基金资助项目（12YJ01）
