中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2011年
11期
887-891
,共5页
结直肠肿瘤%人类皮层肌动蛋白基因%RNA干扰%慢病毒载体
結直腸腫瘤%人類皮層肌動蛋白基因%RNA榦擾%慢病毒載體
결직장종류%인류피층기동단백기인%RNA간우%만병독재체
Colorectal neoplasms%EMS1%RNA interference%Lentiviral vector
目的 探讨通过慢病毒载体介导的人类皮层肌动蛋白基因( EMSl) siRNA沉默皮层肌动蛋白( Cortactin)的表达对结肠癌细胞侵袭能力的影响.方法 构建针对Cortactin的编码基因EMS1的siRNA慢病毒载体转染结肠癌细胞HCT8,分为3组:EMS1-siRNA转染组、空载体组,以及未转染组.Western blot检测3组Cortactin蛋白的表达；采用Transwell小室,观察3组癌细胞的侵袭能力的变化.取4～6周龄的裸鼠共30只,随机分成3组,每组10只.通过尾静脉分别注射上述3种细胞,6周后处死并观察这3组动物的肺转移灶形成情况.结果 EMS1 -siRNA转染细胞的Cortactin表达以及其侵袭能力较另外两组细胞明显降低(P＜0.05).转染组、空载体组和未转染组裸鼠的肺转移率分别为20％、80％和80％,转染组肺转移率明显下降(P＜0.05).结论 Cortactin的表达减少可以降低结肠癌细胞的侵袭能力,并能降低肿瘤在动物体内器官转移的能力.
目的 探討通過慢病毒載體介導的人類皮層肌動蛋白基因( EMSl) siRNA沉默皮層肌動蛋白( Cortactin)的錶達對結腸癌細胞侵襲能力的影響.方法 構建針對Cortactin的編碼基因EMS1的siRNA慢病毒載體轉染結腸癌細胞HCT8,分為3組:EMS1-siRNA轉染組、空載體組,以及未轉染組.Western blot檢測3組Cortactin蛋白的錶達；採用Transwell小室,觀察3組癌細胞的侵襲能力的變化.取4～6週齡的裸鼠共30隻,隨機分成3組,每組10隻.通過尾靜脈分彆註射上述3種細胞,6週後處死併觀察這3組動物的肺轉移竈形成情況.結果 EMS1 -siRNA轉染細胞的Cortactin錶達以及其侵襲能力較另外兩組細胞明顯降低(P＜0.05).轉染組、空載體組和未轉染組裸鼠的肺轉移率分彆為20％、80％和80％,轉染組肺轉移率明顯下降(P＜0.05).結論 Cortactin的錶達減少可以降低結腸癌細胞的侵襲能力,併能降低腫瘤在動物體內器官轉移的能力.
목적 탐토통과만병독재체개도적인류피층기동단백기인( EMSl) siRNA침묵피층기동단백( Cortactin)적표체대결장암세포침습능력적영향.방법 구건침대Cortactin적편마기인EMS1적siRNA만병독재체전염결장암세포HCT8,분위3조:EMS1-siRNA전염조、공재체조,이급미전염조.Western blot검측3조Cortactin단백적표체；채용Transwell소실,관찰3조암세포적침습능력적변화.취4～6주령적라서공30지,수궤분성3조,매조10지.통과미정맥분별주사상술3충세포,6주후처사병관찰저3조동물적폐전이조형성정황.결과 EMS1 -siRNA전염세포적Cortactin표체이급기침습능력교령외량조세포명현강저(P＜0.05).전염조、공재체조화미전염조라서적폐전이솔분별위20％、80％화80％,전염조폐전이솔명현하강(P＜0.05).결론 Cortactin적표체감소가이강저결장암세포적침습능력,병능강저종류재동물체내기관전이적능력.
Objective To explore the effect of Cortactin on invasion and metastasis of human colorectal cancer cells through silencing the expression of Cortactin by transfecting colorectal cancer cells with EMS1-siRNA encoded by a recombinant lentiviral vector.Methods EMS1-siRNA lentiviral vector and negative control lentiviral vector were both transfected into HCT8 cells after virus packing,then the expression of Cortaction of the cells was examined by Western blot to determine the inhibited level of Cortactin.The cells were defined as three groups:EMS1-siRNA transfecting cells,negative control transfecting cells and untransfecting cells.Cell invasion was evaluated with Transwell body.In addition,cells metastasis was observed in animal models.Thirty nude mice were evenly divided into three groups and all the mice were injected through tail vein with 1×107 cells/0.2 ml per animal for each group with the three kinds of cells.The mice were sacrificed 6 weeks after injection and examined for lung metastases development.Results The expression of Cortactin and the ability of invasion of the EMS1-siRNA transfecting cells were all decreased.Lung metastasis rates of EMS1-siRNA transfeting group,negative control group and untransfecting group were 20％,80％ and 80％,respectively.Conclusion siRNA silencing of Cortactin expression can decrease the colorectal cancer cells invasion,and can also decrease the cells' implantation metastasis in animals.
