摘要
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.
出处
《徐州医学院学报》
CAS
2017年第3期169-173,共5页
Acta Academiae Medicinae Xuzhou
基金
徐州市科技局项目(KC14SH086)