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- 2017
下调尾型同源盒基因-2对结肠癌细胞的侵袭及迁移能力的影响及其机制
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Abstract:
摘要：目的 观察下调尾型同源盒基因-2(CDX2)对结肠癌细胞侵袭迁移能力的影响，并初步探讨CDX2基因在结肠癌发生发展过程中的作用及机制。方法 构建CDX2 RNAi慢病毒载体，并转染人结肠癌SW480、HT29细胞，应用Western blotting及qRT-PCR 技术检测CDX2基因的干扰效率；应用Transwell及划痕实验检测下调CDX2表达对SW480、HT29细胞侵袭、迁移能力的影响；Western blotting及RT-PCR检测下调CDX2表达对SW480、HT29细胞上皮-间质转化(epithelial-mesenchymal transition, EMT)相关基因(E-cadherin、ZEB-1、Vimentin、Twist、Snail)表达的影响。结果 构建的CDX2 RNAi慢病毒载体可有效地抑制SW480、HT29细胞中CDX2基因的表达；转染LV-CDX2-shRNA组与转染空病毒组(LV-NT-shRNA)及空白对照组相比，侵袭、迁移能力显著增强(P<0.05)；且E-cadherin表达下降，ZEB-1、Vimentin、Twist、Snail表达均升高。结论 下调CDX2可诱导EMT发生，从而增强结肠癌细胞的侵袭迁移能力。
ABSTRACT: Objective To observe the effect of down-regulated CDX2 gene on the migration and invasion abilities of colon cancer cells (SW480 and HT29) and investigate the role and mechanisms of CDX2 gene in occurrence and development of colon cancer metastasis. Methods CDX2 gene in HT29 and SW480 cells was down-regulated using lentivirus RNA interference (RNAi) vector. The interference efficiency of CDX2 was detected by qRT-PCR and Western blotting. The effect of down-regulated CDX2 expression on colon cancer cells’ migration and invasion was determined by Transwell and wound heal methods. Then the effects of down-regulated CDX2 on the expressions of epithelial-mesenchymal transition (EMT)-related genes (E-cadherin, ZEB-1, Vimentin, Twist and Snail) were detected by RT-PCR and Western blotting. Results The constructed CDX2 siRNA expression vector could significantly inhibit the expression of CDX2 in HT29 and SW480 cells. Compared with those of the cells transfected with empty vector (LV-NT-shRNA) and non-transfected cells, the migration and invasion abilities of cells transfected with LV-CDX2-shRNA were markedly enhanced (P<0.05). E-cadherin expression was reduced while expressions of ZEB-1, Vimentin, Twist, and Snail were significantly increased (all P<0.05). Conclusion Down-regulating the expression of CDX2 can induce the occurrence of EMT, thus enhancing the invasion and migration of colon cancer cells
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