中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
2期
316-318
,共3页
陆建华%李茂岚%江林%吴向嵩%吴文广%董平%顾钧%毕建威
陸建華%李茂嵐%江林%吳嚮嵩%吳文廣%董平%顧鈞%畢建威
륙건화%리무람%강림%오향숭%오문엄%동평%고균%필건위
胃肿瘤%小干扰RNA%Kruppel样转录因子8
胃腫瘤%小榦擾RNA%Kruppel樣轉錄因子8
위종류%소간우RNA%Kruppel양전록인자8
Gastric neoplasm%Interference RNA%Kruppel like factor 8
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.
目的 觀察小榦擾RNA (siRNA)沉默Kruppel樣轉錄因子8(KLF8)基因對胃癌細胞MKN-28增殖及侵襲能力的影響.方法 構建針對KLF8的小榦擾RNA(si-KLF8),應用實時定量聚閤酶鏈反應(Real-time PCR)和Westem blot技術檢測轉染後胃癌細胞中KLF8 mRNA和蛋白的錶達,應用噻唑藍(MTT)比色法檢測轉染24、48、72 h的細胞增殖能力的變化,應用Transwell小室實驗檢測轉染48 h後胃癌細胞侵襲能力的變化.結果 si-KLF8和陰性對照siRNA (si-CTRL)成功轉染至胃癌細胞;si-KLF8組KLF8 mRNA和蛋白的錶達量明顯低于陰性對照組和正常對照組(P＜0.05);si-KLF8組增殖能力下降,24、48、72 h的吸光度值分彆為0.125±0.003、0.193±0.005、0.223±0.005、0.267 ±0.003、0.315±0.006,穿膜細胞數為(42.0±3.6)箇,均明顯低于陰性對照組和正常對照組(P＜0.05);陰性對照組和正常對照組比較,上述指標差異無統計學意義(P＞0.05).結論 si-KLF8可抑製人胃癌細胞的增殖和侵襲能力,提示KLF8在胃癌的髮生髮展過程中髮揮著重要的作用.
목적 관찰소간우RNA (siRNA)침묵Kruppel양전록인자8(KLF8)기인대위암세포MKN-28증식급침습능력적영향.방법 구건침대KLF8적소간우RNA(si-KLF8),응용실시정량취합매련반응(Real-time PCR)화Westem blot기술검측전염후위암세포중KLF8 mRNA화단백적표체,응용새서람(MTT)비색법검측전염24、48、72 h적세포증식능력적변화,응용Transwell소실실험검측전염48 h후위암세포침습능력적변화.결과 si-KLF8화음성대조siRNA (si-CTRL)성공전염지위암세포;si-KLF8조KLF8 mRNA화단백적표체량명현저우음성대조조화정상대조조(P＜0.05);si-KLF8조증식능력하강,24、48、72 h적흡광도치분별위0.125±0.003、0.193±0.005、0.223±0.005、0.267 ±0.003、0.315±0.006,천막세포수위(42.0±3.6)개,균명현저우음성대조조화정상대조조(P＜0.05);음성대조조화정상대조조비교,상술지표차이무통계학의의(P＞0.05).결론 si-KLF8가억제인위암세포적증식화침습능력,제시KLF8재위암적발생발전과정중발휘착중요적작용.
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.
