scholarly journals Quantifying the Rate, Degree, and Heterogeneity of Morphological Change during an Epithelial to Mesenchymal Transition Using Digital Holographic Cytometry

2020 ◽  
Vol 10 (14) ◽  
pp. 4726
Author(s):  
Sofia Kamlund ◽  
Birgit Janicke ◽  
Kersti Alm ◽  
Robert L. Judson-Torres ◽  
Stina Oredsson
Keyword(s):  
Cell Line ◽  
Morphological Change ◽  
Efficient Method ◽  
Mammalian Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Morphological Changes ◽  
Cell Populations ◽  
Label Free ◽  
Mesenchymal Transition ◽  
Quantitative Changes

Cells in complex organisms can transition between epithelial and mesenchymal phenotypes during both normal and malignant physiological events. These two phenotypes are not binary, but rather describe a spectrum of cell states along an axis. Mammalian cells can undergo dynamic and heterogenous bidirectional interconversions along the epithelial–mesenchymal phenotypic (EMP) spectrum, and such transitions are marked by morphological change. Here, we exploit digital holographic cytometry (DHC) to develop a tractable method for monitoring the degree, kinetics, and heterogeneity of epithelial and mesenchymal phenotypes in adherent mammalian cell populations. First, we demonstrate that the epithelial and mesenchymal states of the same cell line present distinct DHC-derived morphological features. Second, we identify quantitative changes in these features that occur hours after induction of the epithelial to mesenchymal transition (EMT). We apply this approach to achieve label-free tracking of the degree and the rate of EMP transitions. We conclude that DHC is an efficient method to investigate morphological changes during transitions between epithelial and mesenchymal states.

scholarly journals Anti-Cancer Effects of Glaucarubinone in the Hepatocellular Carcinoma Cell Line Huh7 via Regulation of the Epithelial-To-Mesenchymal Transition-Associated Transcription Factor Twist1

2021 ◽  
Vol 22 (4) ◽  
pp. 1700
Author(s):  
Jihye Seo ◽  
Jain Ha ◽  
Eunjeong Kang ◽  
Haelim Yoon ◽  
Sewoong Lee ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Wound Healing ◽  
Transcription Factor ◽  
Cell Migration ◽  
Cell Line ◽  
Intracellular Signaling ◽  
Epithelial To Mesenchymal Transition ◽  
Mesenchymal Transition ◽  
Huh7 Cells ◽  
Anti Cancer

Hepatocellular carcinoma (HCC), the most common type of liver cancer, is a leading cause of cancer-related deaths. As HCC has a high mortality rate and its incidence is increasing worldwide, understanding and treating HCC are crucial for resolving major public health concerns. In the present study, wound healing screening assays were performed using natural product libraries to identify natural chemicals that can inhibit cancer cell migration. Glaucarubinone (GCB) showed a high potential for inhibiting cell migration. The anti-cancer effects of GCB were evaluated using the HCC cell line, Huh7. GCB showed anti-cancer effects, as verified by wound healing, cell migration, invasion, colony formation, and three-dimensional spheroid invasion assays. In addition, cells treated with GCB showed suppressed matrix metalloproteinase activities. Immunoblotting analyses of intracellular signaling pathways revealed that GCB regulated the levels of Twist1, a crucial transcription factor associated with epithelial-to-mesenchymal transition, and mitogen-activated protein kinase. The invasive ability of cancer cells was found to be decreased by the regulation of Twist1 protein levels. Furthermore, GCB downregulated phosphorylation of extracellular signal-regulated kinase. These results indicate that GCB exhibits anti-metastatic properties in Huh7 cells, suggesting that it could be used to treat HCC.

scholarly journals Suitable parameter choice on quantitative morphology of A549 cell in epithelial–mesenchymal transition

2015 ◽  
Vol 35 (3) ◽  
Author(s):  
Zhou-Xin Ren ◽  
Hai-Bin Yu ◽  
Jian-Sheng Li ◽  
Jun-Ling Shen ◽  
Wen-Sen Du
Keyword(s):  
Quantitative Evaluation ◽  
A549 Cell ◽  
Epithelial To Mesenchymal Transition ◽  
Epithelial Mesenchymal Transition ◽  
Morphological Changes ◽  
Radius Ratio ◽  
Quantitative Morphology ◽  
Parameter Choice ◽  
Mesenchymal Transition ◽  
Suitable Parameter

Evaluation of morphological changes in cells is key for study of epithelial to mesenchymal transition. In the paper, parameters for quantitative evaluation of the changes were explored and two suitable parameters were found, roundness and radius ratio.

scholarly journals Emerging Effects of Sepantronium Bromide (YM155) on MOLT-4 Cell Line Apoptosis Induction and Expression of Critical Genes Involved in Apoptotic Pathways

2019 ◽  
Vol 10 (1) ◽  
pp. 81-87
Author(s):  
Kobra Shojaei Moghadam ◽  
Majid Farshdousti Hagh ◽  
Mohammad Reza Alivand ◽  
Masoumeh Fardi ◽  
Ali Akbar Movassaghpour ◽  
...  
Keyword(s):  
Cell Line ◽  
Epithelial To Mesenchymal Transition ◽  
Target Genes ◽  
Lymphoblastic Leukemia ◽  
Annexin V ◽  
Apoptosis Induction ◽  
Mesenchymal Transition ◽  
Expression Levels ◽  
Apoptotic Pathways ◽  
Sepantronium Bromide

Purpose: Sepantronium bromide (YM155) is a Survivin inhibitor which recently advanced as an anticancer agent in phase II clinical trials. Survivin belongs to IAP (inhibitor of apoptosis) gene family and is a pivotal target for treatment due to its overexpression and oncogenic function in many malignancies, including acute lymphoblastic leukemia (ALL). Although survivin is a specific target for YM155, recent reports have shown that it has many other crucial targets that regulate its anti-apoptotic effects. The aim of this study was to investigate whether YM155 could have an effect on cell death-inducing genes as well as inducing apoptosis in T-ALL MOLT4- cell line. Methods: We treated MOLT-4 cells with increasing concentrations of YM155 and then cell viability was determined using MTT (methyl thiazolyl tetrazolium) assay. Also, the rate of induction of apoptosis in MOLT-4 cells and the target genes expression levels were evaluated by Annexin V/PI and real-time PCR, respectively. Results: YM155 inhibited cell growth in MOLT-4 cells. This outcome is achieved by inducing apoptosis and a significant increase in the expression level of P53, MiR-9, caspase 3 and decreasing the mRNA expression levels of survivin, Sirtuin1(SIRT1), member of anti-apoptotic proteins family (Bcl-2), and epithelial-to-mesenchymal transition (EMT) initiating factors Snail1and Zeb2. Conclusion: The results showed that use of YM155 can be a potential drug therapy in T-ALL patients with promising effects on apoptosis induction.

scholarly journals Probing single-cell metabolism reveals prognostic value of highly metabolically active circulating stromal cells in prostate cancer

2020 ◽  
Vol 6 (40) ◽  
pp. eaaz3849
Author(s):  
Francesca Rivello ◽  
Kinga Matuła ◽  
Aigars Piruska ◽  
Minke Smits ◽  
Niven Mehra ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Single Cell ◽  
Cancer Patients ◽  
Stromal Cells ◽  
Metastatic Prostate Cancer ◽  
Epithelial To Mesenchymal Transition ◽  
Therapy Response ◽  
Sequencing Analysis ◽  
Label Free ◽  
Mesenchymal Transition

Despite their important role in metastatic disease, no general method to detect circulating stromal cells (CStCs) exists. Here, we present the Metabolic Assay-Chip (MA-Chip) as a label-free, droplet-based microfluidic approach allowing single-cell extracellular pH measurement for the detection and isolation of highly metabolically active cells (hm-cells) from the tumor microenvironment. Single-cell mRNA-sequencing analysis of the hm-cells from metastatic prostate cancer patients revealed that approximately 10% were canonical EpCAM+ hm-CTCs, 3% were EpCAM− hm-CTCs with up-regulation of prostate-related genes, and 87% were hm-CStCs with profiles characteristic for cancer-associated fibroblasts, mesenchymal stem cells, and endothelial cells. Kaplan-Meier analysis shows that metastatic prostate cancer patients with more than five hm-cells have a significantly poorer survival probability than those with zero to five hm-cells. Thus, prevalence of hm-cells is a prognosticator of poor outcome in prostate cancer, and a potentially predictive and therapy response biomarker for agents cotargeting stromal components and preventing epithelial-to-mesenchymal transition.

Optimization of pCMV3-Nog-C-GFPSpark Electro-transfection in MCF-7, a Breast Cancer Cell Line

2021 ◽  
Vol 16 (10) ◽  
pp. 13-18
Author(s):  
Tehrani Azadeh Aghvami ◽  
Saeid Latifi-Navid ◽  
Saber Zahri ◽  
Mohsen Sagha
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cell Line ◽  
Epithelial To Mesenchymal Transition ◽  
Cancer Cell Line ◽  
Bmp Signaling ◽  
Mesenchymal Transition ◽  
Mcf 7

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.

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