Method of enhancing miR-185 expression to reduce low density lipoprotein/cholesterol accumulation in a cell
The present invention provides a method of quantifying miR-185 as a potential biomarker in lipid disorder or cardiovascular diseases in human. The present invention also provides a method of modulating miR-185 in regulating LDL and cholesterol metabolism in cells. The present invention has therapeutic potential in the treatment of cholesterol/LDL related cardiovascular diseases in humans.
1. A method of treating a human cell in need of reducing LDL and cholesterol accumulation, comprising the steps of:
a) providing a human cell in need of reducing LDL and cholesterol accumulation; and
b) transfecting in said cell a miR-185 precursor to cause an increase in miR-185 expression, said miR-185 precursor consisting of a nucleotide sequence set forth in SEQ ID NO: 1,
wherein said increased miR-185 expression in said cell leads to:
i inhibition of HMGCR,
ii inhibition of squalene synthase, and
iii. reduced expression of LDLR,
thereby reducing LDL and cholesterol accumulation in said cell.
2. The method of claim 1 , wherein said human cell is selected from the group consisting of a liver cell, vascular smooth muscle cell and macrophage.
3. The method of claim 1 , wherein said human cell is a liver cell.
4. The method of claim 1 , wherein said transfecting step is performed using electroporation, DEAE-dextran, calcium phosphate, or cationic liposome.
5. A method of reducing LDL and cholesterol accumulation in a human cell, comprising the steps of:
a) providing a human cell in need of reducing LDL and cholesterol accumulation; and
b) transfecting said cell with a miR-185 precursor so as to increase miR-185 expression in said cell,
wherein said increased miR-185 expression causes a reduction of LDL and
cholesterol accumulation in said human cell.
6. The method of claim 5 , wherein said miR-185 precursor consists of the sequence set forth in SEQ ID NO: 1.
7. The method of claim 5 , wherein said transfecting step is performed using electroporation, DEAE-dextran, calcium phosphate, or cationic liposome.
8. A method for altering the expression level of at least a gene selected from the group consisting of HMGCR, squalene synthase, and LDLR in a cell, comprising the steps of;
a) providing a cell in need of altering the expression level of at least a gene selected from the group consisting of HMGCR gene, squalene synthase gene, and LDLR gene; and
b) transfecting in said cell with a composition comprising a miR-185 precursor, said miR-185 precursor consisting of a nucleotide sequence set forth in SEQ ID NO: 1,
wherein said transfecting step of said miR-185 precursor causes inhibition of HMGCR gene, inhibition of squalene synthase gene, and reduced expression of LDLR gene in said cell.