Bmi-1-siRNA对食管鳞癌EC9706细胞体外侵袭迁移的影响

Effects of Bmi - 1 - siRNA on the invasion and migration of EC9706 cells in vitro

目的研究RNA干扰（RNAi）沉默Bmi-1基因表达对人食管鳞癌EC9706细胞体外侵袭、迁移的影响。方法通过合成针对Bmi-1的小干扰RNA（siRNAi）沉默EC9706细胞中Bmi-1的表达，检测该细胞体外侵袭、迁移能力的变化。设置转染Bmi-1siRNA组、转染对照siRNA组和未转染EC9706细胞的对照组。通过逆转录PCR（RT—PCR）、Westernblotting方法检测人端粒酶逆转录酶（hTERT）、P16在3组细胞中的表达。结果转染Bmi-1siRNA组EC9706细胞中Bmi-1的表达水平显著下调，与其他2组比较，差异具有统计学意义（P〈0．05）。Boyden小室体外侵袭实验结果证实，未转染组和转染对照siRNA组的穿膜细胞明显高于转染Bmi-1siRNA组的穿膜细胞数，差异具有统计学意义（P〈0．05）。下调Bmi-1基因的表达可以降低hTERT表达及提高P16的表达。结论RNAi抑制Bmi-1基因表达能够降低食管鳞癌细胞的体外侵袭、迁移能力，可能与hTERT表达的下调、P16表达的上调有关。

Objective To investigate the effects of silencing Bmi -1 gene by RNA interference （RNAi） on the in- vasion and migration of human esophageal squamous carcinoma EC9706 cells in vitro. Methods The expression of Bmi - 1 gene in EC9706 cells was silenced through small RNA interference （siRNAi） specific to Bmi - 1. The consequent changes of the invasion and migration of EC9706 ceils were investigated. Meanwhile, the resultant cells were divided into the following groups ： a Bmi - 1 siRNA transfection group, a transfection control group and a non - transfection group. Then, the levels of human telomerase reverse transcriptase （hTERT） and P16 in the above three groups were analyzed by RT - PCR and Western blotting. Results The level of Bmi - 1 was markedly down - regulated in EC9706 cells after RNA interference, with significant difference from the other two groups （P 〈 0.05）. Besides, according to Boyden cham- ber results, the number of migrated cells in the transfection control and non - transfection groups was greatly higher than that of the Bmi - 1 siRNA transfection group （P 〈 0.05 ）. Furthermore, silencing of Bmi - 1 gene expression could de- crease the amount of hTERT but increase the level of P16. Conclusions Silencing of Bmi - 1 gene expression by RNAi can reduce the invasion and migration of EC9706 cells, which may be associated with down - regulation of hTERT and up - regulation of P16.