Using siRNA Technology for Intensifying Therapeutic Potency of Methotrexate in Breast Cancer Cells

سال انتشار: 1394
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 442

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شناسه ملی سند علمی:

NASTARANCANSER01_173

تاریخ نمایه سازی: 26 شهریور 1395

چکیده مقاله:

Breast cancer is the most prevalent cancer in women and the second cause of cancer deathafter lung cancer in the word. Mutations in the BRCA1, BRCA2, P53 or PTEN genes havebeen related to the breast cancer. Methotrexate is the antineoplastic folic acid analog that isbind to the enzyme dihydrofolate reductase, prevents the conversion of dihydrofolate totetrahydrofolate, and nowadays is used to treatment of the breast cancer. Today RNAinterference -a double strand RNA that has 21-23 nucleotides length- is an important tool forstudyingof gene function.It can silence geneexpression so, increasesthe pharmacologicaleffects of methotrexate.DFF40/CAD is the latent endonuclease that inhibited byDFF45/iCAD. When DFF45 is silent, DFF40 is released and triggered the apoptotic pathway.In this study, DFF45 was silenced and its cytotoxic effects were evaluated in presence andabsence of methotrexate. MCF-7 cell line was cultured in RPMI-1640 medium and LC50measured with MTT assay. Cells were transfected with lipofectamine RNAi max, RNAextracted with RNX-plus and cDNA synthesized.Expression of DFF40 and DFF45genes ofapoptosis pathway were assessed using Real Time PCR. The results of MTT showed theconcentration of methotrexate 0/2 ÂμM kills 50% of the cells in treatments lasting 48 hours.Real time PCR showed that DFF45was knocked down but theexpression of theDFF40genewas increased. In this study, several treatments were performed on MCF7 cell line thatincludes drug treatment with methotrexate simultaneous with methotrexate siRNA and siRNAtreatment alone. The result indicated that drug and siRNA treatment simultaneously, increaseexpression of DFF40 gene, so this treatment is more efficient than the other treatments. Thisover expression was led to trigger cell death pathways.

نویسندگان

Atefeh Ghobadi

School of Biology, College of Science, University of Tehran,Tehran, Iran

Shahrokh Safarian

School of Biology, College of Science, University of Tehran,Tehran, Iran

Seyed Jala Zargar

School of Biology, College of Science, University of Tehran,Tehran, Iran