Evaluation of the effect of SB203580, camptothecin and metformin on cell viability in human breast cancer cell lines MDA-MB-231 and MCF-7

Azadeh Samiei Sefat, Mahdi Ghatreh Samani ©

Evaluation of the effect of SB203580, camptothecin and metformin on cell viability in human breast cancer cell lines MDA-MB-231 and MCF-7

کد: G-26109

نویسندگان: Azadeh Samiei Sefat, Mahdi Ghatreh Samani ©

زمان بندی: زمان بندی نشده!

برچسب: ایمنی شناسی

دانلود: دانلود پوستر

خلاصه مقاله:

Background and Aim

Breast cancer is the most common malignancy diagnosed in women and leads to death in many countries. Researches has shown that metformin inhibits the proliferation and growth of tumor cells, especially breast tumors, by activation of AMP-activated protein kinase (AMPK). On the other hand, SB203580, as a Mitogen-Activated Protein Kinase-α (P38α) inhibitor, inhibits cell growth and induces apoptosis in the cancer cell line. The use of camptothecin, as a Nuclear Receptor subfamily 4 group A (NR4A) inhibitor, reduces cancer cell invasion and metastasis by inducing failure in single-stranded DNA, In regard to, there is limited information on the simultaneous effects of metformin, camptothecin and SB203580 on cancer cells, we intended to investigate the single and combined effects of these drugs on the survival rate of breast cancer cell lines.

Methods

The human Breast cancer cell lines MCF7 and MDA-MB-231 were seeded onto 96-well plates in 100 μl of RPMI 1640, which was supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin, at a density of 3 × 〖10〗^3 cells per well for 24 hours before treatment. Then cell treated with metformin (10 mM), SB203580 (10μM) and CPT (25nM) alone and in combination for 24 and 48 hours. The viability of cells was then examined by a colorimetric test that reflects the number of viable cells present. Briefly, 20 µL of MTT (5 mg/mL in PBS) was added to each well and incubated in a dark place at 37˚C for 3 hours. Thereafter evacuating the contents of the plate, 100 μl of DMSO was added to each well and the plates were incubated for additional 20 min. The absorbance was measured by an Elisa reader at 590 nm.

Results

The viability of MDA-MB-231 cells in group of metformin was 60% and 24% after 24 and 48 hours treatment, respectively while in the SB group, respectively, 96% and 86% and in the CPT group 100% and 91%. Combination of metformin/SB decreased cell viability compared to single treatment after 24 and 48 hours but there wasn't significant decrease in combination of metformin/CPT and also in combination of SB/CPT after 24 and 48 hours. Our results indicated that combination of metformin/SB/CPT leads to significant decrease of cell viability after 24 and 48 hours. The viability of MCF7 cells in group of metformin was 76% and 48% after 24 and 48 hours treatment, respectively while in the SB group, respectively, 93% and 104% and in the CPT group 95% and 91%. There wasn't significant decrease in combination of metformin/CPT and also in combination of SB/CPT after 24 and 48 hours. Our results indicated that combination of metformin/SB/CPT also didn't leads to decrease of cell viability after 24 and 48 hours.

Conclusion

In this study, we observed that metformin decreases MDA-MB-231 cell viability especially after 48 hours incubation while SB and CPT were not able to inhibit cancer cell growth, but in combination with metformin, either as a double combination (with the exception of the Metformin/CPT group, which was seen as a slight decrease due to metformin) or as a triple combination, greatly enhances the effect of metformin. We observed that metformin decreases MCF7 cell viability especially after 48 hours incubation while SB and CPT alone and in combination together even in combination with metformin were not able to inhibit cancer cell growth.

Keywords

Camptothecin; Metformin; P38α MAP kinase; NR4A; SB203580

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