摘要
该文通过RNA干扰方法下调染色体结构维持蛋白3(structural maintenance of chromosomes protein 3,SMC3)的表达,探究其对人肺癌细胞A549的影响。用Western blot验证SMC3敲低的效果,用CCK-8实验、Transwell实验检测SMC3敲低对A549细胞的增殖和迁移能力的影响,用细胞成球等实验检测SMC3敲低对A549细胞干性的影响。结果显示,该研究成功地构建了p SUPERSMC3干扰质粒,将其转染A549细胞后SMC3蛋白质水平明显降低。实验组(p SUPER-SMC3)与正常对照组(control)及阴性对照组(p SUPER)相比,细胞增殖和迁移能力明显下降(P<0.01),细胞成球数减少,干性减弱。该实验结果表明,下调SMC3蛋白质水平可以抑制A549细胞的增殖和迁移能力,减弱细胞干性,提示SMC3对肺癌的发生和发展可能具有促进作用。该实验为寻找潜在的抗肺癌方法提供了新的实验依据。
In this study, the level of SMC3(structural maintenance of chromosomes protein 3) protein was down-regulated by RNA interference, then we explore the effect of SMC3 on human lung cancer cell line A549. The level of SMC3 protein was determined by Western blot. The effect of SMC3 down-regulation on the proliferation and migration of A549 cells was detected by CCK-8 assay and Transwell assay. The effect of SMC3 down-regulation on the stemness of A549 cells was detected by sphere formation. The results showed that the p SUPER-SMC3 interference plasmid was successfully constructed, and the level of SMC3 protein was significantly decreased in A549 cells trasfected with the p SUPER-SMC3. Compared with the control group and the negative control group(p SUPER), the cell proliferation and migration ability of the SMC3-knocked down group(p SUPERSMC3) decreased significantly(P〈0.01), and the number of sphere formation and the stemness decreased, indicating that SMC3 could promote the development and progression of lung cancer and might provide a new experimental basis for finding potential anti-lung cancer methods.
出处
《中国细胞生物学学报》
CAS
CSCD
2017年第10期1320-1325,共6页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:U1302225
81460253
81460417
81473342)
云南省高端科技人才基金(批准号:2012HA008)
云南省教育厅面上项目(批准号:KKJA201426013)
云南省省级自然科学人才培养项目(批准号:KKSY201326121)资助的课题~~