[siRNA-cyclin D1 inhibit cell proliferation in breast cancer MCF-7 cell line].
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.
Jiang
Lei
L
Institute of Cell Biology, Zhejiang University, Hangzhou.
Chen
Ri Shu
RS
Li
Ji Cheng
JC
D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D005260:Female; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D034741:RNA, Small Interfering; D019938:Cyclin D1