论文部分内容阅读
背景与目的:有丝分裂检测点缺陷可引起细胞染色体不稳定性而使细胞生物学行为改变,本实验探讨RNA干扰有丝分裂阻滞缺陷蛋白2(mitotic arrest deficient2,MAD2)基因表达对人食管鳞癌细胞系KYSE30细胞增殖和侵袭能力的影响。方法:利用脂质体转染的方法,将MAD2基因的siRNA转染食管鳞癌细胞KYSE30,并通过RT-PCR以及免疫印迹的方法进行siRNA效果鉴定。实验分为正常对照组、非特异干扰组、特异干扰组。MTT、克隆形成实验检测细胞增殖活性的变化,损伤刮擦实验和Transwell小室实验分别检测转染细胞株的迁移与侵袭能力。结果:siRNA作用48h组,MAD2蛋白的表达最低;相应地,迁移黏附能力增强,侵袭实验显示转染后细胞侵袭能力较转染前有显著加强。结论:MAD2基因沉默能够促进人食管鳞癌细胞系KYSE30的体外增殖和侵袭能力增强,并抑制凋亡,提示其变化对肿瘤的发生和转移发挥重要作用。
BACKGROUND & AIM: The mitotic arrest deficient2 (MAD2) gene expression in human esophageal squamous cell carcinoma cell line (ESCC) is affected by mitotic checkpoint defects that cause cell chromosome instability and cell biological behavior. KYSE30 cell proliferation and invasion ability. Methods: siRNA targeting MAD2 gene was transfected into KYSE30 cells by lipofection. The effect of siRNA was identified by RT-PCR and Western blotting. The experiment was divided into normal control group, non-specific interference group, specific interference group. MTT and clonogenic assay were used to detect the changes of cell proliferation activity. Scratch-scratching assay and Transwell chamber assay were used to detect the migration and invasion ability of transfected cell lines respectively. Results: The expression of MAD2 protein was the lowest in 48 hours after siRNA treatment. Correspondingly, the ability of migration and adhesion was enhanced. The invasion assay showed that the invasive ability of cells after transfection was significantly enhanced compared with that before transfection. CONCLUSION: MAD2 gene silencing can promote the proliferation and invasion of human esophageal squamous cell carcinoma cell line KYSE30 in vitro and inhibit its apoptosis, suggesting that the changes play an important role in tumorigenesis and metastasis.