scholarly journals Increased vimentin mRNA expression in MCF-7 breast cancer cell line after repeated endoxifen-treatment

2017 ◽  
Vol 25 (4) ◽  
pp. 207-13 ◽  
Author(s):  
Paramita Paramita ◽  
Melva Louisa ◽  
Nafrialdi Nafrialdi

Background: Epithelial mesenchymal transition (EMT) plays a significant role in the development of cancer cell resistance to drugs. Vimentin, a type III intermediate filament protein, is a marker of EMT. Vimentin's over-expression in cancer correlates well with increased tumor growth, change in cell shape and poor prognosis. Endoxifen is an active metabolite of tamoxifen  and has become a new potent agent in the treatment of breast cancer. This is a study that aimed to investigate the effect of endoxifen exposure with or without estradiol on cell viability, cell morphology and EMT progression through the analysis of vimentin mRNA expression after 4-week treatment. Methods: Endoxifen, 100 nM or 1,000 nM, with or without beta-estradiol were given repeatedly to MCF-7 cells. Cells treated with dimethyl sulfoxide (DMSO) 0.001% were used as control. After 2- and 4-week exposure, the cells were counted, analyzed for mRNA vimentin expression, and observed for morphological changes. Results: Compared to control, there were significant decreases in vimentin mRNA expressions in endoxifen and endoxifen+β-estradiol treated cells after 2-weeks, which then significantly increased after 4-week compared with the 2-week exposure. We found no change in morphology of MCF-7 cells. Conclusion: Repeated exposure of endoxifen might induce EMT progression through increased expression of vimentin in MCF-7 breast cancer cell line.

2021 ◽  
Vol 16 (10) ◽  
pp. 13-18
Author(s):  
Tehrani Azadeh Aghvami ◽  
Saeid Latifi-Navid ◽  
Saber Zahri ◽  
Mohsen Sagha

Bone morphogenetic protein (BMP) signaling is known as one of the most important pathways in breast cancer tumorigenesis. This triggers the epithelial to mesenchymal transition (EMT) in metastatic cells and consequently the migrating cells become invasive and noggin, a BMP antagonist prevents it. So, the present study aimed to optimize Noggin transfection into MCF-7 as a breast cancer cell line. Following DH5α bacterial cell culturing and pCMV3- Nog-GFPSpark transformation, the resulted plasmid was extracted, purified and finally transfected into MCF-7 at different voltages (100-230V), resistances (1000 and ∞ Ω) and capacitances (25-75μF) using the electroporation system with various concentrations of plasmid (between 30 and 100μg/ml). As a result, we found that noggin has a better transfection into MCF- 7 in 230V, 50μF, 1000 Ωof electroporator. At 80μg/ml concentration of plasmid, the cell expressing GFP also represented the noggin expression.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Stefania Nobili ◽  
Antonella Mannini ◽  
Astrid Parenti ◽  
Chiara Raggi ◽  
Andrea Lapucci ◽  
...  

AbstractInvasive ductal carcinoma (IDC) constitutes the most frequent malignant cancer endangering women’s health. In this study, a new spontaneously immortalized breast cancer cell line, DHSF-BR16 cells, was isolated from the primary IDC of a 74-years old female patient, treated with neoadjuvant chemotherapy and disease-free 5-years after adjuvant chemotherapy. Primary breast cancer tissue surgically removed was classified as ER−/PR−/HER2+, and the same phenotype was maintained by DHSF-BR16 cells. We examined DHSF-BR16 cell morphology and relevant biological and molecular markers, as well as their response to anticancer drugs commonly used for breast cancer treatment. MCF-7 cells were used for comparison purposes. The DHSF-BR16 cells showed the ability to form spheroids and migrate. Furthermore, DHSF-BR16 cells showed a mixed stemness phenotype (i.e. CD44+/CD24−/low), high levels of cytokeratin 7, moderate levels of cytokeratin 8 and 18, EpCAM and E-Cadh. Transcriptome analysis showed 2071 differentially expressed genes between DHSF-BR16 and MCF-7 cells (logFC > 2, p-adj < 0.01). Several genes were highly upregulated or downregulated in the new cell line (log2 scale fold change magnitude within − 9.6 to + 12.13). A spontaneous immortalization signature, mainly represented by extracellular exosomes-, plasma membrane- and endoplasmic reticulum membrane pathways (GO database) as well as by metabolic pathways (KEGG database) was observed in DHSF-BR16 cells. Also, these cells were more resistant to anthracyclines compared with MCF-7 cells. Overall, DHSF-BR16 cell line represents a relevant model useful to investigate cancer biology, to identify both novel prognostic and drug response predictive biomarkers as well as to assess new therapeutic strategies.


2021 ◽  
Vol 66 (1) ◽  
pp. 65-71
Author(s):  
Hung Nguyen Phi ◽  
Toan Tran Quoc ◽  
Tuan Nguyen Anh ◽  
Vy Trinh Ngoc Thao ◽  
Yen Ngo Thi Ngoc ◽  
...  

Three alkaloids (named rutaecarpine (1), evodiamine (2), schinifoline (3) and one phenylpropanoid, integrifoliodiol (4)) have been isolated from the EtOAc extract of the fruits of Tetradium ruticarpum (A. Juss.) T. G. Hartley collected in Lang Son province. Their structures have been identified by using 1D and 2D NMR spectroscopies. All four compounds were tested for their cytotoxicity against the human breast cancer cell line (MCF-7) and tamoxifen-resistant breast cancer cell line (MCF\TAMR). The results showed that rutaecarpine (1) inhibited the growth of MCF7 and MCF\TAMR with its IC50 values of 41.2 and 64.6 µM, respectively. In addition, compounds 1, 2, and 4 showed moderate activity toward MCF-7 cell line.


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