长链非编码 RNA ZEB1-AS1 通过调节上皮性卵巢癌细胞中的 MMP19 影响紫杉醇和顺铂耐药性。
Long noncoding RNA ZEB1-AS1 affects paclitaxel and cisplatin resistance by regulating MMP19 in epithelial ovarian cancer cells.
机构信息
Nanjing Maternal and Child Health Institute, Women's Hospital of Nanjing Medical University (Nanjing Maternity and Child Health Care Hospital), Nanjing, 210004, China.
Department of Gynecology, Women's Hospital of Nanjing Medical University (Nanjing Maternity and Child Health Care Hospital), Nanjing, 210004, China.
出版信息
Arch Gynecol Obstet. 2021 May;303(5):1271-1281. doi: 10.1007/s00404-020-05858-y. Epub 2020 Nov 5.
PURPOSE
The long noncoding RNA (lncRNA) ZEB1-AS1 is reported overexpressed in sensitive ovarian cancer cells A2780 compared with paclitaxel (PTX)-and cisplatin (DDP)- resistant. However, the function and mechanism of ZEB1-AS1 in EOC cells still unknown.
METHODS
We used quantitative real-time PCR (qPCR) to detect ZEB1-AS1 expression in A2780 and A2780/R cells. A combination of siRNA, plasmids, CCK8 and flow cytometry was used to detect the effect of ZEB1-AS1 on ovarian cancer cell A2780 PTX and DDP resistance. Transcriptome sequencing, qPCR, and western blot were used for further mechanistic studies.
RESULTS
ZEB1-AS1 depletion using siRNA in chemosensitive A2780 cells significantly increased PTX and DDP resistance. In contrast, ZEB1-AS1 overexpression in PTX- and DDP-resistant A2780/resistant (A2780/R) cells reversed the observed drug resistance. Thus, ZEB1-AS1 plays an important role in PTX and DDP resistance in EOC cells. However, quantitative real-time PCR (qPCR) and western blot results suggested that ZEB1-AS1 did not regulate chemoresistance through regulation of ZEB1 protein. We used sequencing to detect mRNA expression changes in A2780 cells after ZEB1-AS1 silencing. The results indicated that MMP19 was the likely downstream factor of ZEB1-AS1. We further examined whether ZEB1-AS1 played an important role in chemoresistance by silencing MMP19 in ZEB1-AS1-overexpressing cells. CCK8 assay results suggested that MMP19 knockdown promoted ZEB1-AS1-induced chemoresistance to PTX and DDP in A2780 cells.
CONCLUSION
This study is the first to reveal that ZEB1-AS1 plays a pivotal role in cancer chemoresistance.
目的
长链非编码 RNA（lncRNA）ZEB1-AS1 在敏感卵巢癌细胞 A2780 中的表达高于紫杉醇（PTX）和顺铂（DDP）耐药细胞。然而，ZEB1-AS1 在卵巢癌细胞中的功能和机制尚不清楚。
方法
我们使用实时定量 PCR（qPCR）检测 A2780 和 A2780/R 细胞中 ZEB1-AS1 的表达。采用 siRNA、质粒、CCK8 和流式细胞术组合检测 ZEB1-AS1 对卵巢癌细胞 A2780 PTX 和 DDP 耐药性的影响。转录组测序、qPCR 和 Western blot 用于进一步的机制研究。
结果
用 siRNA 敲低化疗敏感的 A2780 细胞中的 ZEB1-AS1 显著增加了 PTX 和 DDP 的耐药性。相反，在 PTX 和 DDP 耐药的 A2780/耐药（A2780/R）细胞中过表达 ZEB1-AS1 逆转了观察到的耐药性。因此，ZEB1-AS1 在卵巢癌细胞的 PTX 和 DDP 耐药中起重要作用。然而，实时定量 PCR（qPCR）和 Western blot 结果表明，ZEB1-AS1 并未通过调节 ZEB1 蛋白来调节化疗耐药性。我们使用测序检测 ZEB1-AS1 沉默后 A2780 细胞中 mRNA 表达的变化。结果表明，MMP19 可能是 ZEB1-AS1 的下游因子。我们进一步研究了在 ZEB1-AS1 过表达细胞中沉默 MMP19 是否对 ZEB1-AS1 引起的 PTX 和 DDP 化疗耐药性发挥重要作用。CCK8 检测结果表明，MMP19 敲低促进了 ZEB1-AS1 诱导的 A2780 细胞对 PTX 和 DDP 的化疗耐药性。
结论
本研究首次揭示 ZEB1-AS1 在癌症化疗耐药中起关键作用。
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