【摘要】 目的:研究酪氨酸激酶受体RON过表达对结肠癌细胞移动/浸润能力的影响。方法:将携有野生型RON(wt-RON)cDNA的质粒pDR2-wt-RON转染入结肠癌细胞株RKO,挑选稳定转染克隆,以过河实验和体外跨室趋化运动能力实验检测两者的移动/浸润能力;然后以siRNA敲除,比较两者的移动/浸润能力,以Western印迹检测E-cadherin表达的变化。结果:转染了wt-RON并且高表达后,RKO细胞的趋化移动能力明显高于未转染组(P<0.01)。过河实验,转染组过河时间为(42.50±4.12)h,而未转染组与载体对照组分别为(69.50±2.52)h与(70.50±3.42)h(P<0.01);而基因干扰后,趋化移动能力降低(P<0.01)。过河实验,RNAi组过河时间为(82.50±3.42)h,与未转染组(79.00±2.58)h相仿(P>0.05),psiRm-RON组(51.50±4.12)h(P<0.01)。转染wt-RON后,E-cadherin表达低于未转染组(P<0.05)。结论:wt-RON的高表达可以降低E-cadherin表达,降低肿瘤细胞间的黏附性,增加结肠癌细胞RKO的移动/浸润能力。实施RNAi(RNA interference)可降低RKO的移动/浸润能力。提示RON酪氨酸激酶受体的高表达可能是结肠癌浸润转移的机制之一。更多还原

【Abstract】 AIM: To investigate the roles of overexpression of RON receptor tyrosine kinase in motile/invasive ability of human colorectal cancer cell line RKO.METHODS: A eucaryotic expression vector pDR2 containing full-length wt-RON cDNA was transfected into the colorectal cancer cell line RKO and a stable expression clone was obtained.The motile/invasive ability was tested by wound healing test and the transwell migration assay.The expression of E-cadherin was measured by Western blotting.RESULTS: Motile ability of transfected RKO was greatly promoted by transwell chemotaxis assay(P<0.01).The wound healing time showed statistical difference as of(42.50±4.12) h,(69.50±2.52) h and(70.50±3.42) h,respectively in transfected group,untransfected group and vector control group.After knocking down RON by siRNA,the motile became less than that in control group(P<0.01).E-cadherin expression in transfected RKO was decreased significantly due to pDR2-wt-RON transfection.CONCLUSIONS: Overexpression of wt-RON led to the decrease in expression of E-cadherin and decreased cancer cell-cell adhension.At the same time, migration/invasion ability was promoted.Taken together,abnormal accumulation of RON might play potential roles in invasion/metastasis of colorectal cancer.RNAi can block motile/invasion ability mediated by RON.更多还原