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KMID : 0353320140380020073
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Oral Biology Research
2014 Volume.38 No. 2 p.73 ~ p.82
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Analysis of microRNA expression in KB human oral cancer cells
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Mo Shin-Yeob
Kim Jeong-Sun
Cho Seon-Ho
Park Jong-Tae
Yu Sun-Kyuong
Kim Do-Kyung
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Abstract
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Purpose: The main aim of this study was to compare and analyze expression profiles of miRNAs to establish miRNA-related cancer cell growth inhibition in normal human oral keratinocytes (NHOK) and KB human oral cancer cells.
Materials and Methods: Expression profiles of miRNAs in NHOK and KB cells were examined by miRNA microarray analysis, quantitative real-time polymerase chain reaction analysis (qRT-PCR), 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and gene array analysis.
Results: In the miRNA microarray analysis, 164 and 149 miRNAs were up- and down-regulated, respectively, in KB cells compared to NHOK among 1,769 miRNAs examined. miR-30a, miR-99a, and miR-155 were up-regulated more than 10-fold in KB cells compared to NHOK, whereas miR-205, miR-203, and miR-200c were down-regulated more than 10-fold. In qRT-PCR analysis, expression levels of miR-30a, miR-99a, and miR-155 increased in KB cells compared to NHOK more than 15-fold, whereas miR-205, miR-203, and miR-200c were down-regulated more than 10-fold. Importantly, overexpression of miR-205 and miR-203 significantly inhibited growth of KB cells. In gene array analysis, 3,154 and 2,709 genes were up- and down-regulated more than 2-fold, respectively, in miR-205-overexpressing KB cells compared to control KB cells. Overexpressed miR-203 in KB cells induced expression of 2,707 genes and decreased that of 2,352 genes.
Conclusion: These results show that miR-205 and miR-203 expression was reduced in KB cells compared to NHOK, and proliferation of KB human oral cancer cells was inhibited. Moreover, these in vitro results indicate that miR-205 and miR-203 have significant therapeutic potential as molecular medicine for treatment of oral cancer by turning on silenced tumor suppressor genes through miRNA targeting.
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KEYWORD
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Cancer therapy, Cell death, Human oral keratinocytes, KB human oral cancer cells, MicroRNAs
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