论文部分内容阅读
目的:探讨通过封闭ERCC1基因表达干扰DNA损伤修复路径及其对卵巢癌细胞对铂类药物耐药性的影响。方法:构建ERCC1基因反义表达载体,转染人卵巢癌细胞A2780及A2780/CF70,采用Northern杂交及Western Blot分析人卵巢癌细胞内ERCC1基因RNA及蛋白表达水平变化;利用荧光素酶报告基因系统观察宿主细胞对DNA损伤的修复作用;采用MTT实验研究细胞对铂类药物耐药性的影响。结果:卵巢癌细胞转染反义ERCC1基因后,ERCC1基因转录水平、蛋白表达水平明显下降。荧光素酶报告基因系统结果证明,宿主细胞的DNA修复活性下降。MTT实验表明,伴随ERCC1基因表达水平下降,细胞对顺铂的耐药性降低。结论:转染ERCC1反义基因显著影响宿主细胞的核苷酸切除修复,影响细胞对顺铂的耐药性。
OBJECTIVE: To investigate the effect of blocking ERCC1 gene expression on DNA damage repair pathways and its resistance to platinum drugs in ovarian cancer cells. Methods: The antisense expression vector of ERCC1 gene was constructed and transfected into human ovarian cancer cells A2780 and A2780 / CF70. Northern blotting and Western Blot were used to analyze the expression of ERCC1 mRNA and protein in human ovarian cancer cells. The luciferase reporter gene system To observe the repair effect of host cells on DNA damage; MTT assay was used to study the effect of cells on the drug resistance of platinum drugs. Results: After transfection with antisense ERCC1 gene in ovarian cancer cells, ERCC1 gene transcription and protein expression were significantly decreased. The luciferase reporter gene system results demonstrate that DNA repair activity of the host cell is decreased. MTT experiments show that, accompanied by decreased expression of ERCC1 gene, cell resistance to cisplatin decreased. CONCLUSION: Transfection of ERCC1 antisense gene significantly affects nucleotide excision and repair of host cells and affects cell resistance to cisplatin.