Lv Teng, Song Kejuan, Zhang Lili, Li Weihua, Chen Yulong, Diao Yuchao, Yao Qin, Liu Peishu
a Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, Shandong, China.
b Department of Obstetrics and Gynecology, The Affiliated Hospital of Qingdao University, Qingdao, China.
Biochem Cell Biol. 2018 Oct;96(5):663-671. doi: 10.1139/bcb-2018-0031. Epub 2018 Mar 21.
This study aimed to explore the roles of miRNA-34a (miR-34a) in ovarian cancer (OC) cells and uncover possible mechanisms. The proliferation of OC cells was measured with an MTT assay and soft agar colony formation assay. TargetScan analysis, real-time PCR, and a luciferase reporter assay were used to demonstrate the downstream target of miR-34a in OC cells. HDAC1 expression levels were detected by immunoblot analysis. miR-34a inhibited the proliferation of SKOV3 and OVCA433 cells and enhanced cisplatin sensitivity in cisplatin-resistant SKOV3cp cells. The results of TargetScan analysis, real-time PCR, and luciferase reporter assay confirmed that miR-34a downregulated HDAC1 expression by directly targeting the 3'-UTR of HDAC1 mRNA. The overexpression of HDAC1 decreased cisplatin sensitivity and promoted proliferation in OC cells. MTT assay and soft agar colony formation assay showed that HDAC1 overexpression blocked the suppressive effects of miR-34a on SKOV3 cell proliferation. In addition, treatment with the miR-34a mimic partially recovered the cisplatin sensitivity of SKOV3cp cells, whereas HDAC1 overexpression blocked the above phenomena caused by treatment with the miR-34a mimic. miR-34a exhibited suppressive effects on OC cells via directly binding and downregulating HDAC1 expression, which subsequently decreased the resistance to cisplatin and suppressed proliferation in OC cells.
本研究旨在探讨微小RNA-34a（miR-34a）在卵巢癌细胞（OC）中的作用，并揭示其可能的机制。采用MTT法和软琼脂集落形成试验检测OC细胞的增殖情况。运用TargetScan分析、实时定量聚合酶链反应（real-time PCR）和荧光素酶报告基因试验来证实miR-34a在OC细胞中的下游靶点。通过免疫印迹分析检测组蛋白去乙酰化酶1（HDAC1）的表达水平。miR-34a抑制了SKOV3和OVCA433细胞的增殖，并增强了顺铂耐药的SKOV3cp细胞对顺铂的敏感性。TargetScan分析、实时定量聚合酶链反应和荧光素酶报告基因试验的结果证实，miR-34a通过直接靶向HDAC1 mRNA的3'-非翻译区（3'-UTR）下调HDAC1的表达。HDAC1的过表达降低了OC细胞对顺铂的敏感性并促进了其增殖。MTT法和软琼脂集落形成试验表明，HDAC1的过表达阻断了miR-34a对SKOV3细胞增殖的抑制作用。此外，用miR-34a模拟物处理可部分恢复SKOV3cp细胞对顺铂的敏感性，而HDAC1的过表达则阻断了用miR-34a模拟物处理所引起的上述现象。miR-34a通过直接结合并下调HDAC1的表达对OC细胞发挥抑制作用，进而降低OC细胞对顺铂的耐药性并抑制其增殖。
