RNA干扰沉默神经型钙黏素基因对人食管 癌细胞系EC9706侵袭能力的影响
Effect of RNA Mediated Gene Silencing of N-cadherin Expression on Invasiveness of EC9706
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摘要: 探讨RNAi沉默N-cadherin基因表达对人食管癌细胞系EC9706体外侵袭能力的影响。方法 将N-cadherin基因的RNA干扰载体pMSCVneo/N-cadherin质粒通过脂质体转染法转染人食管癌细胞系EC9706，运用G418进行抗性克隆的筛选和扩增，进而得到稳定转染的人食管癌细胞系EC9706，通过RT-PCR和Western blot检测稳定转染后的人食管癌细胞系EC9706中N-cadherin的mRNA和蛋白表达水平的变化，同时检测转染前后食管癌细胞系EC9706中MMP-9基因表达水平的变化，并通过Transwell小室体外侵袭实验检测转染前后人食管癌细胞系EC9706体外侵袭能力的变化。结果 通过G418加压筛选法得到稳定转染的人食管癌细胞系EC9706，RT-PCR和Western blot检测结果显示，稳定转染后的人食管癌细胞系EC9706中N-cadherin的mRNA和蛋白表达水平均下调，同时MMP-9基因的mRNA表达水平也随之下调；Transwell小室体外侵袭实验结果显示，伴随着N-cadherin和MMP-9表达水平的下调，人食管癌细胞系EC9706的体外侵袭能力也降低（P＜0.05）。结论 RNA干扰沉默神经型钙黏素基因可以使人食管癌细胞系EC9706的体外侵袭能力降低。Abstract: Objective To investigate cell invasive power of human EC9706 cell line when the N-cadherin expression was down-regulated by RNA interference (RNAi) technology. Methods N- cadherin shRNA was transfected into EC9706 cells to inhibit the expression of N-cadherin. The effect of RNAi was detected by RT-PCR and Western blot. The invasive ability of the cancer cells was determined by Transwell assay. Results After RNAi，the expression of N-cadherin was significantly decreased, which indicated that RNAi was effective. With the decreased expression of N-cadherin and MMP-9，cell invasiveness was dramatically depressed. Conclusion The down-regulation of N-cadherin protein could impair the invasivness of EC9706 cell line.
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