Objective To preliminarily explore the role of long non-coding RNA(lncRNA)HOX transcriptional antisense intergenic RNAs(HOTAIR)in the malignant transformation of liver cells induced by microcystin-LR(MCLR).Methods WRL68 cells were continuously exposed to 10 nmol/L MCLR to the 25th passage,and the malignancy of the cells was determined by soft agar clone formation assay.The 25th passage of MCLR infected cells and the passage-matched control cells were harvested for lncRNA microarray analysis,observed the expression of HOTAIR,and bioinformatics database was utilized to predict the target genes and possible interacting miRNAs of HOTAIR.The expression levels of HOTAIR and its target genes were detected by real-time fluorescence quantitative PCR(qRT-PCR)in control group and infected group cells at passage 0(P0),5(P5),10(P10),15(P15),20(P20),and 25(P25).Results The number of cell clones formed at P25 infected group was higher than the control group of the same passage.lncRNA microarray screening showed that the expression of HOTAIR in the P25 infected group was upregulated by 2.471-fold,and the expression of cyclin-dependent kinase inhibitor 1A(CDKN1A),a target gene of HOTAIR,was upregulated by 2.733-fold in MCLR-induced cells of P25 compared to the control group.qRT-PCR results showed that the expression of HOTAIR was upregulated by 2.411-fold,2.566-fold,and 3.183-fold,respectively,and the CDKN1A mRNA expression was upregulated by 1.579-fold,1.864-fold,and 3.351-fold in MCLR infected cells of P15,P20,P25,respectively,compared to the passage-matched control group.At each stage of MCLR-induced malignant transformation,the expression level of miR-17-5p,a microRNA commonly found in HOTAIR and CDKN1A,gradually decreased with the increasing number of passages of MCLR exposure.Overexpression of miR-17-5p could inhibit the up-regulation of HOTAIR and CDKN1A induced by chronic MCLR exposure.Conclusions HOTAIR and CDKN1A showed sustained high expressions,whereas miR-17-5p showed consistently low expression during MCLR-induced malignant transformation of liver cells.HOTAIR may regulate each other with miR-17-5p,and subsequently affect the expression of the target gene CDKN1A.
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