筛选KIAA1456抑制卵巢癌细胞增殖的靶向lncRNAs

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

目的筛选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-q PCR和Western blot检测KIAA1456干扰效率。用RT-q PCR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。

Objective Screening the potential lncRNAs of KIAA1456 inhibiting ovarian cancer cells. Methods Immunofluorescence was used to detect the expression of KIAA1456 in HO8910 PM ovarian cancer cell. Microarray was performed to identify differently expressed lncRNAs in HO8910 PM ovarian cancer cells overexpressed KIAA1456 as compared with the normal control group. The interesting candidate lncRNAs were verified by realtime quantitative PCR. Constructing RNAi lentiviral vector targeting KIAA1456 gene in HO8910 PM ovarian cancer cells,RT-q PCR and Western blot was performed to measure the interference efficiency. Differentially expressed lncRNAs were validated by RT-q PCR after KIAA1456 knock-down. Results KIAA1456 mainly expressed in nucleus of HO8910 PM 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 lncRNAsincluding down-regulated gene SSX8 and up-regulated genes SNORD14 D,SNORA26,lncRNA-ENST00000384488.The results of the RT-q PCR were consistent with the data from the microarray. The shRNA interference vectors targeting the KIAA1456 gene were successfully constructed,SSX8 was up-regulated,and SNORD14 D,SNORA26,lncRNA-ENST00000384488 were down-regulated after knockdown the expression of KIAA1456 in the HO8910/PM ovarian cancer cells. Conclusions KIAA1456 inhibits proliferation of ovarian cancer by regulating correlative lncRNAs,which may provide a new target for the diagnosis and treatment of ovarian cancer.