Study on the regulation mechanism of long noncoding RNA on dentin matrix protein 1

doi:10.3877/cma.j.issn.1674-1366.2017.01.002

Abstract

Abstract:

Objective

To investigate the regulation mechanism of long noncoding RNA (lncRNA) on dentin matrix protein 1 (DMP1) .

Methods

The lipidosome infection protocol was used to transfect the lncRNA32865 plasmid and lncRNA32865 siRNA into the MC3T3-E1 cells. Using the methods of Western blot and RT-qPCR to detect the expression tendency of lncRNA32865 and DMP1 after transfection. The method of luciferase reporter gene was used to examine the DMP1 promoter activity and the effect of lncRNA32865 on it. For all statistical analyses, P<0.05 was considered statistically significant.

Results

Over expression of lncRNA32865 led to the decrease of DMP1 expression (0.236 ± 0.022) , there was a statistically significant difference between the groups (t= 59.816, P<0.001) . DMP1 expression was increased (1.994 ± 0.133) caused by inhibiting lncRNA32865 gene, there was a statistically significant difference between the groups (t=-12.989, P= 0.006) . The luciferase reporter gene analysis showed that DMP1 promoter was active (t=-77.360, P<0.001) . The Luciferase expression level of DMP1 promotor and lncRNA32865 plasmid cotransfection (3.877 ± 0.120) was lower than the control group (6.018 ± 0.105) with statistically significant difference (t= 50.713, P<0.001) . The Luciferase expression level of DMP1 promotor and lncRNA32865 siRNA cotransfection was increased (17.296 ± 0.674) with statistically significant difference (t=-26.612, P= 0.001) .

Conclusions

Western blot and RT-qPCR analyses demonstrated that the expression tendency of lncRNA32865 and DMP1 is opposite. LncRNA32865 regulates the expression of DMP1 gene by binding to the DMP1 promoter.

Key words: Gene expression regulation, RNA untranslated, Long chain, Dentin matrix proteins

Boya Li, Hong Wu, Xin Xia, Jiangyuan Chen, Yansong Yu, Peilin Zhuang, Yi Ruan. Study on the regulation mechanism of long noncoding RNA on dentin matrix protein 1[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2017, 11(01): 7-11.

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Figures/Tables 5

References 16

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基因	引物序列
DMP1	F:5′-TGGTATCAGGTCGGAAGAATC-3′
?	R:5′-CCCTGCTGTTGCTGTCAGTA-3′
lncRNA32865	F:5′-CTCAGCATCTCTGGTAAGGT-3′
?	R:5′-AGGTAGGGCATGGACTAGAA-3′
GAPDH	F:5′-GGCCTCCAAGGAGTAAGAAA-3′
?	R:5′GCCCCTCCTGTTATTATGG-3′
U6	F:5′CTCGCTTCGGCAGCACA-3′
?	R:5′AACGCTTCACGAATTTGCGT-3′

基因	引物序列
DMP1	F:5′-TGGTATCAGGTCGGAAGAATC-3′
?	R:5′-CCCTGCTGTTGCTGTCAGTA-3′
lncRNA32865	F:5′-CTCAGCATCTCTGGTAAGGT-3′
?	R:5′-AGGTAGGGCATGGACTAGAA-3′
GAPDH	F:5′-GGCCTCCAAGGAGTAAGAAA-3′
?	R:5′GCCCCTCCTGTTATTATGG-3′
U6	F:5′CTCGCTTCGGCAGCACA-3′
?	R:5′AACGCTTCACGAATTTGCGT-3′

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