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目的:探究Chk1反义寡核苷酸(CHK1-ASODN)单独或联合顺铂(DDP)对卵巢癌细胞系SKOV-3侵袭转移能力的影响,并阐明其可能的分子机制。方法:体外培养人卵巢癌细胞系SKOV-3,CHK1-ASODN单独或联合DDP处理48 h后,划痕实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力;显微镜下观察细胞上皮或间质表型特征;Western blot及实时定量PCR技术分别检测上皮间质转化(EMT)特异性标志物(E-cadherin、N-cadherin)以及EMT关键调控分子ZEB1的蛋白及m RNA的表达水平。结果:与对照组相比较,CHK1-ASODN单独或联合DDP均能显著抑制SKOV-3细胞的迁移及侵袭(P<0.05);细胞表现为间质化表型;E-cadherin的表达显著升高(P<0.05),而N-cadherin的表达则显著降低(P<0.05);ZEB1的表达显著降低(P<0.05)。结论:CHK1-ASODN单独或联合DDP下调ZEB1的表达进而逆转EMT可能是其抑制卵巢癌侵袭转移的重要机制之一。
Objective: To investigate the effect of Chk1 antisense oligodeoxynucleotides (CHK1-ASODN) alone or in combination with cisplatin (DDP) on the invasion and metastasis of ovarian cancer cell line SKOV-3 and elucidate its possible molecular mechanism. Methods: The migration of human ovarian cancer cell line SKOV-3, CHK1-ASODN alone or in combination with DDP for 48 h was observed by scratch test. The cell invasion was evaluated by Transwell assay. The epithelial or mesenchymal phenotype Western blot and real-time quantitative PCR were used to detect the expression of E-cadherin (N-cadherin) and ZEB1 protein and m RNA in EMT. Results: Compared with the control group, CHK1-ASODN alone or in combination with DDP significantly inhibited the migration and invasion of SKOV-3 cells (P <0.05); the cells showed interstitial phenotype and the expression of E-cadherin was significantly increased (P <0.05), while the expression of N-cadherin was significantly lower (P <0.05); the expression of ZEB1 was significantly lower (P <0.05). Conclusion: The down-regulation of ZEB1 expression by CHK1-ASODN alone or in combination with DDP, which may be one of the important mechanisms of its inhibition on the invasion and metastasis of ovarian cancer.