| Purpose:The aim was to explore the expression of SATB1(the Special A-T rich sequence binding protein) in breast cancer cells. A lentiviral expression vector for RNA interference (RNAi) of SATB1 gene was constructed,then its gene silencing effect in highly aggressive human breast cancer cell line MDA-MB-231 was assessed.Methods:1.The expression of SATB1 mRNA in breast cancer cells (MDA-MB-231) was observed by Real-time fluorescent quantitative PCR. 2. The expression of SATB1 in breast cancer cells was observed by Western-blotting. 3.Single-strand DNA oligos which was SATB1-targeting were designed and synthesized.Then the lentivirus vector which contained RNAi cassette was used to transduct into MDA-MB-231. In the last part, SATB1-targeting RNAi lentivirus inhibited the expression of SATB1 gene in MDA-MB-231 was observed by Real-time fluorescent quantitative PCR.And its silencing effect on proliferation in MDA-MB-231 was observed by MTT assay.Results:1. SATB1 mRNA was expressed in aggressive breast cancer cells (MDA-MB-231). 2.The expression of SATB1 had higher level in aggressive breast cancer cells (MDA-MB-231) than normal human mammary epithelial cells (HMEC) .3. A lentiviral vector for RNA interference of SATB1 gene was successfully constructed,and the sequence and correct site of ds oligos inserted were confirmed by PCR and sequencing assay.4.The results of Real-time fluorescent quantitative PCR showed that the expression of SATB1 in MDA- MB-231 could be inhibited efficiently by RNAi.The results of MTT assay suggested that inhibiting the expression of SATB1 by RNAi could down-regluate the proliferation of MDA-MB-231 efficiently.Conclusion:1.The SATB1-targeting RNAi lentiviral vector was an important tool that can inhibit the expression of SATB1 gene efficiently.2. Inhibiting the expression of SATB1 down-regluate the proliferation of breast cancer cells which implied that SATB1 might promote the metastasis of breast cancer .
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