[siRNA-cyclin D1 inhibit cell proliferation in breast cancer MCF-7 cell line].
Lei Jiang, Ri Shu Chen, Ji Cheng Li
Abstract
Lei Jiang, Ri Shu Chen, Ji Cheng Li
Abstract
To study the inhibitory effect of siRNA on cyclin D1 expression and cell proliferation in breast cancer MCF-7 cell line. The siRNA targeting cyclin D1 was chemically synthesized and transfected into MCF-7 cells by oligofectamine. The expression of cyclin D1 was analyzed by quantitive PCR and Western blot, and the cell growth inhibition was measured with CCK-8 assay. Then, cell cycle of the transfected cells was examined by flow cytometry, and cell colony forming ability was measured by soft-agar colony formation assay. After MCF-7 cells were transfected with 10, 50, 100nmol/L siRNA,the expression of cyclin D1 mRNA was respectively suppressed with inhibition rates of 57.85%, 63.22% and 68.02%, and the protein expression was suppressed with inhibition rates of 51.13%, 62.09% and 77.68% respectively. The proliferation of MCF-7 cells was inhibited after transfection with siRNA-cyclin D1, which caused cell cycle arrest at G1 phase and showed less colony forming ability in the breast cancer cell line MCF-7. These results indicate that siRNA-cyclin D1 could be a powerful anti-proliferative tool in breast cancer gene therapy.
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To study the inhibitory effect of siRNA on cyclin D1 expression and cell proliferation in breast cancer MCF-7 cell line. The siRNA targeting cyclin D1 was chemically synthesized and transfected into MCF-7 cells by oligofectamine. The expression of cyclin D1 was analyzed by quantitive PCR and Western blot, and the cell growth inhibition was measured with CCK-8 assay. Then, cell cycle of the transfected cells was examined by flow cytometry, and cell colony forming ability was measured by soft-agar colony formation assay. After MCF-7 cells were transfected with 10, 50, 100nmol/L siRNA,the expression of cyclin D1 mRNA was respectively suppressed with inhibition rates of 57.85%, 63.22% and 68.02%, and the protein expression was suppressed with inhibition rates of 51.13%, 62.09% and 77.68% respectively. The proliferation of MCF-7 cells was inhibited after transfection with siRNA-cyclin D1, which caused cell cycle arrest at G1 phase and showed less colony forming ability in the breast cancer cell line MCF-7. These results indicate that siRNA-cyclin D1 could be a powerful anti-proliferative tool in breast cancer gene therapy.
Key concepts: Cyclin D1, MCF-7, Cell cycle, Transfection, Cell growth, Cyclin D, Cyclin, Molecular biology