Chk1反义寡核苷酸联合顺铂抑制卵巢癌侵袭转移的分子机制

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(P0.05);细胞表现为间质化表型;E-cadherin的表达显著升高(P0.05),而N-cadherin的表达则显著降低(P0.05);ZEB1的表达显著降低(P0.05)。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。

The Molecular Mechanisms Underlying CHK1-ASODN Alone or Combined with DDP Inhibits Human Ovarian Cancer Cells Migration and Invasion

Objective:To explore the effects of CHK1-ASODN alone or combined with DDP on human ovarian cancer cells migration and invasion, and investigate the molecular mechanisms underlying these effects.Methods:Human ovarian cancer cell line SKOV-3 was cultured in vitro, and then treated with CHK1-ASODN alone or combined DDP for 48 h. The wound healing assay was performed to detect SKOV-3 cell migration. The invasion capacity of SKOV-3 cell was determined in vitro using transwell assay. The morphological changes of SKOV-3 cell were detected by microscope. The expression levels of E-cadherin, N-cadherin and ZEB1 protein were detected by Western blot and real time PCR.Results:Treatment of human ovarian cancer cell line SKOV-3 with CHK1-ASODN alone or combined with DDP resulted in significant inhibition of cell migration and invasion (P < 0.05). The results ofWestern blot and RT-PCR showed that E-cadherin expression was upregulated and the expression of N-cadherin and ZEB1 was downregulated.Conclusion:CHK1-ASODN alone or combined with DDP can inhibit ovarian cancer cell invasion and migration, which may be through down-regulating ZEB1expression, then reversing the EMT.