复旦学报(医学版)
複旦學報(醫學版)
복단학보(의학판)
JOURNAL OF FUDAN UNIVERSITY
2010年
2期
202-206
,共5页
钟锡明%范钰%周永静%陈坚%林庚金
鐘錫明%範鈺%週永靜%陳堅%林庚金
종석명%범옥%주영정%진견%림경금
大肠肿瘤%肝转移%畸胎瘤衍生生长因子-1%小干扰RNA
大腸腫瘤%肝轉移%畸胎瘤衍生生長因子-1%小榦擾RNA
대장종류%간전이%기태류연생생장인자-1%소간우RNA
colorectal carcinoma%liver metastasis%teratocarcinoma-derived growth factor-1%small interfering RNA
目的 研究cripto基因对人大肠癌细胞迁移、侵袭和肝转移的影响.方法 以cripto基因mRNA小干扰RNA(small interfering RNA,siRNA)转染人大肠癌SW480细胞后,分别采用荧光实时定量RT-PCR和Western blot检测cripto基因mRNA和蛋白水平,以划痕试验方法评测癌细胞迁移能力;用Boyden模型观察癌细胞侵袭能力.以脾切除法建立裸鼠大肠癌肝转移模型,以cripto siRNA转染48 h后的癌细胞接种裸鼠,观察对大肠癌细胞体内肝转移的影响.结果 siRNA转染组cripto基因mRNA水平明显降低,且呈浓度和时间依赖性(P<0.001;P<0.001).体外实验发现,cripto siRNA转染组癌细胞迁移距离和穿膜细胞数目明显低于对照组,且呈浓度依赖性(P<0.05).体内实验发现,cripto基因siRNA转染组SW480细胞转移率和肿瘤结节数均明显低于对照组(P<0.05;P<0.05).结论 cripto基因在大肠癌肝转移中起着重要的作用;以siRNA下调cripto基因表达,可抑制大肠癌细胞肝转移.
目的 研究cripto基因對人大腸癌細胞遷移、侵襲和肝轉移的影響.方法 以cripto基因mRNA小榦擾RNA(small interfering RNA,siRNA)轉染人大腸癌SW480細胞後,分彆採用熒光實時定量RT-PCR和Western blot檢測cripto基因mRNA和蛋白水平,以劃痕試驗方法評測癌細胞遷移能力;用Boyden模型觀察癌細胞侵襲能力.以脾切除法建立裸鼠大腸癌肝轉移模型,以cripto siRNA轉染48 h後的癌細胞接種裸鼠,觀察對大腸癌細胞體內肝轉移的影響.結果 siRNA轉染組cripto基因mRNA水平明顯降低,且呈濃度和時間依賴性(P<0.001;P<0.001).體外實驗髮現,cripto siRNA轉染組癌細胞遷移距離和穿膜細胞數目明顯低于對照組,且呈濃度依賴性(P<0.05).體內實驗髮現,cripto基因siRNA轉染組SW480細胞轉移率和腫瘤結節數均明顯低于對照組(P<0.05;P<0.05).結論 cripto基因在大腸癌肝轉移中起著重要的作用;以siRNA下調cripto基因錶達,可抑製大腸癌細胞肝轉移.
목적 연구cripto기인대인대장암세포천이、침습화간전이적영향.방법 이cripto기인mRNA소간우RNA(small interfering RNA,siRNA)전염인대장암SW480세포후,분별채용형광실시정량RT-PCR화Western blot검측cripto기인mRNA화단백수평,이화흔시험방법평측암세포천이능력;용Boyden모형관찰암세포침습능력.이비절제법건립라서대장암간전이모형,이cripto siRNA전염48 h후적암세포접충라서,관찰대대장암세포체내간전이적영향.결과 siRNA전염조cripto기인mRNA수평명현강저,차정농도화시간의뢰성(P<0.001;P<0.001).체외실험발현,cripto siRNA전염조암세포천이거리화천막세포수목명현저우대조조,차정농도의뢰성(P<0.05).체내실험발현,cripto기인siRNA전염조SW480세포전이솔화종류결절수균명현저우대조조(P<0.05;P<0.05).결론 cripto기인재대장암간전이중기착중요적작용;이siRNA하조cripto기인표체,가억제대장암세포간전이.
Objective To study the effects of cripto on migration, invasion, and liver metastasis of colorectal cancer cell. Methods After human colorectal cancer cell line SW480 was transfected by cripto small interfering RNA (siRNA), the mRNA and protein level were determined by Real-time RT-PCR and Western blot, respectively. The migration and invasion ability were evaluated by wound-healing assay and boyden chamber model, respectively. Thirty nude mice model of liver metastasis from colorectal cancer was established by splenectomy. Results The siRNA could down-regulate the level of mRNA and protein of cripto in a dose- and time-dependent manner. Suppression of cripto expression could inhibit migration and invasion ability of human colorectal cancer cell in vitro. The metastastic rate and tumor nodules were lower in transfection with cripto siRNA than in two control groups in vivo. Conclusions Cripto gene might play an important role in regulation of liver metastasis from colorectal carcinoma cell, and suppression of cripto gene by siRNA can inhibit liver metastasis of colorectal cancer.
