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Synthesized small interfering RNA inhibits the expression of DNA methyltransferase 1 in salivary adenoid cystic carcinoma / 中华口腔医学杂志
Chinese Journal of Stomatology ; (12): 225-228, 2007.
Article in Chinese | WPRIM | ID: wpr-333358
ABSTRACT
<p><b>OBJECTIVE</b>To select the effective siRNA which could inhibit the expression of DNA methyltransferase 1 (DNMT-1) in adenoid cystic carcinoma (ACC) and discuss the time-, and dose-dependent effect of RNA interference (RNAi).</p><p><b>METHODS</b>Four pairs of siRNA were designed, synthesized and transfected through oligofectamine reagent into ACC cell lines ACC-2, ACC-3 and ACC-M. 24, 48 and 72 h after transfection, total RNA and protein were harvested respectively. mRNA and protein expression level of DNMT-1 were detected by real time PCR, RT-PCR and Western blot, and then the effective siRNA was subsequently selected. ACC-3 as also transfected by different concentration of siRNA and the dose-dependent effect of RNAi was discussed. Cy(3) labelled GAPDH siRNA was used as a positive control.</p><p><b>RESULTS</b>Two of 4 pairs of siRNA inhibited the mRNA expression of DNMT-1 in three ACC cell lines and the expression of DNMT-1 was downregulated by 67%, 86%, 92% and 76%, 76%, 86% respectively. The gene inhibition was detected at 24 h or 48 h after transfection, maintained only 1 - 2 days and showed direct relationship to the concentration of siRNA. The change of protein expression level of DNMT-1 was consistent to the changes of mRNA.</p><p><b>CONCLUSIONS</b>The effective siRNA which could inhibited the expression of DNMT-1 of ACC were achieved. The inhibition effect of RNAi was time-dependent and dose-dependent.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: RNA, Messenger / Salivary Gland Neoplasms / Transfection / Carcinoma, Adenoid Cystic / DNA Methylation / RNA, Small Interfering / Cell Line, Tumor / DNA (Cytosine-5-)-Methyltransferases / Genetic Vectors / Genetics Limits: Humans Language: Chinese Journal: Chinese Journal of Stomatology Year: 2007 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: RNA, Messenger / Salivary Gland Neoplasms / Transfection / Carcinoma, Adenoid Cystic / DNA Methylation / RNA, Small Interfering / Cell Line, Tumor / DNA (Cytosine-5-)-Methyltransferases / Genetic Vectors / Genetics Limits: Humans Language: Chinese Journal: Chinese Journal of Stomatology Year: 2007 Type: Article