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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。
Objective: To investigate the effect of tyrosine kinase receptor RON overexpression on colon cancer cell migration / infiltration. METHODS: The plasmid pDR2-wt-RON carrying the wild-type RON (wt-RON) cDNA was transfected into RKO colon cancer cell line. Stable transfected clones were selected and tested by trans-river and in vitro transmembrane chemotaxis The migration / invasion ability of the two proteins was detected by Western blotting. The expression of E-cadherin was detected by siRNA knockdown. Results: After transfected with wt-RON, the chemotactic ability of RKO cells was significantly higher than that of untransfected cells (P <0.01). The river crossing time was (42.50 ± 4.12) h in the transfection group compared with (69.50 ± 2.52) h and (70.50 ± 3.42) h in the untransfected group and the control group (P <0.01) After interference, the chemotactic ability to move decreased (P <0.01). In the river crossing experiment, the time of crossing the river in the RNAi group was (82.50 ± 3.42) h, which was similar to that in the non-transfected group (79.00 ± 2.58) h (P> 0.05) . The expression of E-cadherin in transfected wt-RON group was lower than that in untransfected group (P <0.05). Conclusion: High expression of wt-RON can decrease the expression of E-cadherin, reduce the adhesion between tumor cells and increase the migration / invasion ability of RKO in colon cancer cells. RNA interference can reduce the mobility / infiltration of RKO. Tip RON tyrosine kinase receptor high expression may be one of the mechanisms of invasion and metastasis of colon cancer.