4 research outputs found
Gallic acid and curcumin induce cytotoxicity and apoptosis in human breast cancer cell MDA-MB-231
Introduction: Gallic acid (GA) and curcumin (Cur) are natural phenolic compounds that their anti-tumor effects on many types of cancers have been proved. In the current study, the effect of the combination of these agents on MDA-MB-231 breast cancer cells was investigated. Methods: Inhibition of cell proliferation (MTT assay), light microscopy, fluorescence microscopy, cell cycle analysis, nitrite detection, ROS levels, measurement of the mitochondrial membrane potential, GSH level, Annexin V assay, RT-PCR and Western blotting methods were applied. Results: The results revealed the combination of GA and Cur strongly decreased MDA-MB-231 cell growth. Moreover, this combination increased ROS level and cytotoxic activity along with the glutathione depletion in MDA-MB-231 cells. Flow cytometry analysis showed the combination of GA and Cur increased sub-G1 cell population. Furthermore, fluorescent staining and Annexin V/PI assay showed that apoptotic cells were significantly increased in the presence of GA and Cur. At last, protein expression evaluation showed that the combination of GA and Cur significantly decreased Bcl-2 level while increased Bax, cleaved-caspase3 and PARP levels in MDA-MB-231 cells. Conclusion: These results suggest that GA in combination with Cur could be a possible candidate for chemoprevention agent of triple negative breast cancer
The effect of glycolysis inhibitor dichloroacetate on the apoptosis rate and alteration of gene and miRNA expression of breast cancer cells MDA-MB-231
Breast cancer is the most common cause of death from cancer among women. The triple-negative breast cancer (TNBC) is the most invasive subtype, and chemotherapy is the only therapy option. Cancer cells preferably utilize the glycolysis pathway even with proper oxygen availability, and this activation plays a great role in tumorigenesis. Therefore, glycolysis targeting can be an effective strategy for cancer treatment. Here, the apoptotic effect of a glycolysis inhibitor named dichloroacetate (DCA) on TNBC cells MDA-MB-231 was assessed, and the expression of anti-apoptotic genes and oncogenic miRNAs was evaluated. MTT assay showed that DCA reduces cell viability in a dose-dependent manner with the IC50 concentration of 50 mM. Annexin/PI assay demonstrated that DCA due to DCA treatment. Finally, the expression of anti-apoptotic genes Bcl2l1 and Mcl1 and oncogenic miRNAs miR21 and miR27a decreased due to DCA treatment. Our results confirmed that DCA, as a glycolysis inhibitor, leads to apoptosis induction in TNBC cells because of reducing expression of viability genes and miRNAs
Generation of Rat Embryonic Germ Cells via Inhibition of TGFß and MEK Pathways
Objective: Embryonic germ (EG) cells are the results of reprogramming primordial
germ cells (PGC) in vitro. Studying these cells can be of benefit in determining the
mechanism by which specialized cells acquire pluripotency. Therefore in the current
study we have tried to derive rat EG cells with inhibition of transforming growth
factor-β (TGFβ) and mitogen-activated protein kinase kinase (MEK) signaling pathways.
Materials and Methods: In this experimental study, rat PGCs were cultured under
feeder free condition with two small molecules that inhibited the above mentioned
pathways. Under this condition only two-day presence of stem cell factor (SCF) as a
survival factor was applied for PGC reprogramming. Pluripotency of the resultant EG
cells were further confirmed by immunofluorescent staining, directed differentiation
ability to neural and cardiac cells, and their contribution to teratoma formation as well.
Moreover, chromosomal stability of two different EG cells were assessed through Gbanding
technique.
Results: Formerly, derivation of rat EG cells were observed solely in the presence of glycogen
synthase kinase-3 (GSK3β) and MEK pathway inhibitors. Due to some drawbacks
of inhibiting GSK3β molecules such as increases in chromosomal aberrations, in the present
study we have attempted to assess a feeder-free protocol that derives EG cells by
the simultaneous suppression of TGFβ signaling and the MEK pathway. We have shown
that rat EG cells could be generated in the presence of two inhibitors that suppressed the
above mentioned pathways. Of note, inhibition of TGFβ instead of GSK3β significantly
maintained chromosomal integrity. The resultant EG cells demonstrated the hallmarks of
pluripotency in protein expression level and also showed in vivo and in vitro differentiation
capacities.
Conclusion: Rat EG cells with higher karyotype stability establish from PGCs by inhibiting
TGFβ and MEK signaling pathways