基础医学与临床 ›› 2017, Vol. 37 ›› Issue (4): 518-524.

• 研究论文 • 上一篇    下一篇

筛选KIAA1456抑制卵巢癌细胞增殖的靶向lncRNAs

张应凤1,高燕洪1,陈怀梅2,王佳1   

  1. 1. 重庆医科大学,附属大学城医院
    2. 重庆市急救中心
  • 收稿日期:2016-11-18 修回日期:2016-12-21 出版日期:2017-04-05 发布日期:2017-03-24
  • 通讯作者: 王佳 E-mail:18084014036@163.com
  • 基金资助:
    重庆市渝中区科技计划项目基础与前沿

Screening of potential lncRNAs of KIAA1456 inhibiting proliferation of ovarian cancer cells

  • Received:2016-11-18 Revised:2016-12-21 Online:2017-04-05 Published:2017-03-24

摘要: 目的 探索KIAA1456抑制卵巢癌细胞增殖的靶向lncRNAs。方法 细胞免疫荧光检测KIAA1456在卵巢癌细胞HO8910PM中的表达。用基因芯片技术检测过表达KIAA1456后的卵巢癌细胞HO8910PM,与对照HO8910PM卵巢癌细胞之间差异lncRNA基因表达谱,筛选出有明显差异的lncRNAs,并验证。构建KIAA1456-RNAi慢病毒载体,干扰卵巢癌细胞HO8910/PM中KIAA1456的表达,RT-qPCR和Western blot检测KIAA1456干扰效率。用RT-qPCR再次检测KIAA1456沉默后相关lncRNAs的表达变化。 结果 KIAA1456主要表达于HO8910PM细胞的细胞核。过表达KIAA1456后,表达相差1.2倍以上的lncRNA有654条,表现最显著的有下调的SSX8,上调的SNORD14D,SNORA26,lncRNA-ENST00000384488,PCR验证结果与芯片趋势一致。KIAA1456-RNAi慢病毒载体成功干扰了卵巢癌细胞HO8910PM中KIAA1456的表达,KIAA1456下调后,SSX8上调,SNORD14D,SNORA26,lncRNA-ENST00000384488下调。结论 KIAA1456可通过调节SSX8,SNORD14D,SNORA26,lncRNA-ENST00000384488等相关lncRNAs的表达,从而抑制卵巢癌细胞的增殖,为提高卵巢癌的诊断和治疗提供了一个新的靶点。

关键词: KIAA1456, 基因芯片, 慢病毒载体, RNA干扰, lncRNAs

Abstract: Objective Exploring the potential lncRNAs of KIAA1456 inhibiting ovarian cancer cells . Methods Immunofluorescence was used to detect the expression of KIAA1456 in HO8910PM ovarian cancer cell. Microarray was performed to identify differently expressed lncRNAs in HO8910PM ovarian cancer cells overexpressed KIAA1456 compared with the normal control group. The interesting candidate lncRNAs were verified by real-time quantitative PCR.Constructing RNAi lentiviral vector targeting KIAA1456 gene in HO8910PM ovarian cancer cells, RT-qPCR and Western blot was performed to measure the interference efficiency. Differentially expressed lncRNAs were validated by RT-qPCR after KIAA1456 knock-down. Results KIAA1456 mainly expressed in nucleus of HO8910PM cell. A total of 654 differently expressed lncRNAs were identified in the KIAA1456-overexpressed group compared with the control group with a set filter fold-change ≥1.2, the most aberrantly expressed lncRNAs including down-regulated gene SSX8 and up-regulated genes SNORD14D,SNORA26,lncRNA-ENST00000384488. The results of the RT-qPCR were consistent with the data from the microarray.The shRNA interference vectors targeting the KIAA1456 gene were successfully constructed, SSX8 was up-regulated, and SNORD14D,SNORA26,lncRNA-ENST00000384488 were down-regulated after knockdown the expression of KIAA1456 in the HO8910/PM ovarian cancer cells. Conclusions KIAA1456 could inhibit the proliferation of ovarian cancer by regulating correlative lncRNAs, such as SSX8,SNORD14D,SNORA26,lncRNA-ENST00000384488, which could provide a new target for the diagnosis and treatment of ovarian cancer.

Key words: KIAA1456, Microarray, Lentiviral vector, RNA interference, lncRNAs