Reduction of Rapid Proliferating Tumour Cell Lines by Inhibition of the Specific Glycine Transporter GLYT1

Studies have highlighted the relevance of extracellular glycine and serine in supporting high growth rates of rapidly proliferating tumours. The present study analysed the role of the specific glycine transporter GLYT1 in supplying glycine to cancer cells and maintaining cell proliferation. GLYT1 knockdown in the rapidly proliferating tumour cell lines A549 and HT29 reduced the number of viable cells by approximately 30% and the replication rate presented a decrease of about 50% when compared to cells transfected with control siRNA. In contrast, when compared to control, GLYT1 siRNA had only a minimal effect on cell number of the slowly proliferating tumour cell line A498, reducing the number of viable cells by 7% and no significant difference was observed when analysing the replication rate between GLYT1 knockdown and control group. When utilising a specific GLYT1 inhibitor, ALX-5407, the doubling time of rapidly proliferating cells increased by about 8 h presenting a significant reduction in the number of viable cells after 96 h treatment when compared to untreated cells. Therefore, these results suggest that GLYT1 is required to maintain high proliferation rates in rapidly proliferating cancer cells and encourage further investigation of GLYT1 as a possible target in a novel therapeutic approach.

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TAT-Apoptin Mediated Induction of Apoptosis in Leukaemic Cells.

Blood ◽

10.1182/blood.v108.11.1900.1900 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1900-1900

Author(s):

Joop Gaken ◽

Louisa Pericleous ◽

Farzin Farzaneh ◽

Ghulam J. Mufti ◽

Mahvash Tavassoli

Keyword(s):

Epithelial Cells ◽

Cancer Cells ◽

Cell Lines ◽

Tumour Cell ◽

Solid Tumour ◽

Tumour Cells ◽

Normal Cells ◽

Leukaemic Cells ◽

Tumour Cell Lines

Abstract We have studied the specific targeting of leukaemic cells using the Chicken Anaemia Virus (CAV)-derived protein VP3 (Apoptin) linked to the protein transduction domain (PTD) from HIV TAT with the aim of using this strategy for in vitro purging. Apoptin is a 13.6 kDa protein which induces apoptosis specifically in cancer cells whilst leaving normal cells unaffected. Expression of Apoptin in normal cells results in its cytoplasmic localisation. In tumour cells Apoptin resides initially in the cytoplasm and subsequently translocates to the nucleus and induces apoptosis. Apoptin is phosphorylated both in vitro and in vivo in tumour cells but negligibly in normal cells at threonine 108. A gain-of-function point mutation (T108E) results in accumulation of Apoptin in the nucleus and the killing of normal cells, implying that phosphorylation is a key factor of the tumour-specific properties of Apoptin. We have demonstrated that Apoptin induces apoptosis in a variety of human solid tumour cell lines, but not in normal fibroblast and epithelial cells. Apoptin induced apoptosis in HSC3 head and neck cancer cells acts through the mitochondrial pathway and was blocked (>75%) by shRNA against PUMA, a BH3 Only protein which induces Bax and BAK resulting in loss of mitochondrial membrane potential and release of cytochrome C. Furthermore, activation of the p53 family member, p73, substantially increased (5–10 fold for p73 β and γ) sensitivity of Saos2 tumour cells to Apoptin-induced killing. For efficient protein delivery, Apoptin was fused to a TAT PTD and addition of this protein to normal and tumour cells resulted in the selective killing of tumour cells. To increase the stability and solubility of TAT-Apoptin we have fused it to the maltose binding protein (MBP), this modification significantly increases both yield and the solubility of Apoptin while retaining its biological function. Apoptin tumour specific toxicity was assessed in a range of leukaemic and solid tumour cell lines. Addition of MBP-TAT-Apoptin protein to HL60, K562 and Jurkat cells resulted in 50%, 55% and 75% cell death by apoptosis as judged by PARP cleavage, respectively, at day 4 as compared to MBP-TAT control whilst normal B cells, fibroblasts and epithelial cells are unaffected. Fluorescent microscopy demonstrated that MBP-TAT-Apoptin was rapidly internalised in almost 100% of cells within 24hrs in all cell types tested. Direct injection of Apoptin expressing Ad vectors also showed clear regression of established tumours in mice. The cancer specific toxicity of Apoptin has potential value for a range of therapeutic applications such as purging of autologous bone marrow as used for the treatment of multiple myeloma and possibly direct treatment of leukaemias either alone or linked to antibodies for targeting of specific types of leukaemias.

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The Cytotoxic Effect of Magainin II on the MDA-MB-231 and M14K Tumour Cell Lines

BioMed Research International ◽

10.1155/2013/831709 ◽

2013 ◽

Vol 2013 ◽

pp. 1-11 ◽

Cited By ~ 9

Author(s):

Radu Anghel ◽

Daniela Jitaru ◽

Laurenţiu Bădescu ◽

Magda Bădescu ◽

Manuela Ciocoiu

Keyword(s):

Cell Lines ◽

Tumour Cell ◽

Chemotherapeutic Agents ◽

Breast Adenocarcinoma ◽

Public Health Issue ◽

Tumour Cell Line ◽

Global Public Health ◽

Tumour Cell Lines ◽

Magainin Ii

Many studies have highlighted the tumoricidal properties of some natural peptides known to have antimicrobial virtues. Also, the increasingly higher resistance to conventional antibiotics has become a global public health issue, and the need for new antibiotics has stimulated interest in finding and synthesizing new antimicrobial peptides, which may also be used as chemotherapeutic agents. Relying on the literature, the purpose of ourin vitroresearch was to assess the tumoricidal potential of magainin II on a series of tumour cell lines, namely, MDA-MB-231 (breast adenocarcinoma) and M14K (human mesothelioma). The experimental results of our study revealed that the cytotoxic effects of magainin II depend on its concentration. Its efficiency is significant at 120 μM concentrations, and, although it is much lower, it persists even at 60 μM concentrations. The effects were insignificant at 30 μM concentrations. In our experimental research, the tumoricidal effect of magainin II was not significantly dependent on the type of tumour cell line used.

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Screening of Venezuelan medicinal plant extracts for cytostatic and cytotoxic activity against tumour cell lines

Planta Medica ◽

10.1055/s-0032-1320482 ◽

2012 ◽

Vol 78 (11) ◽

Author(s):

P Taylor ◽

M Arsenak ◽

MJ Abad ◽

Á Fernández ◽

R Gonto ◽

...

Keyword(s):

Medicinal Plant ◽

Cytotoxic Activity ◽

Cell Lines ◽

Plant Extracts ◽

Tumour Cell ◽

Medicinal Plant Extracts ◽

Tumour Cell Lines

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Cytotoxic activity against tumour cell lines and anti-inflammatory effects of compounds isolated from Xylopia aromatica

Planta Medica ◽

10.1055/s-0033-1352067 ◽

2013 ◽

Vol 79 (13) ◽

Author(s):

O Estrada ◽

L González ◽

M Mijares ◽

Á Fernández ◽

M Ruiz ◽

...

Keyword(s):

Cytotoxic Activity ◽

Cell Lines ◽

Tumour Cell ◽

Tumour Cell Lines ◽

Anti Inflammatory

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Epigenetic modifiers reduce proliferation of human neuroendocrine tumour cell lines

Endocrine Abstracts ◽

10.1530/endoabs.31.p149 ◽

2013 ◽

pp. 1-1

Author(s):

E Kate Lines ◽

U Katherine Gaynor ◽

Mark Stevenson ◽

J Paul Newey ◽

E Sian Piret ◽

...

Keyword(s):

Cell Lines ◽

Tumour Cell ◽

Neuroendocrine Tumour ◽

Epigenetic Modifiers ◽

Tumour Cell Lines

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Sevoflurane represses the migration and invasion of gastric cancer cells by regulating forkhead box protein 3

Journal of International Medical Research ◽

10.1177/03000605211005936 ◽

2021 ◽

Vol 49 (4) ◽

pp. 030006052110059

Author(s):

Fangfang Yong ◽

Hemei Wang ◽

Chao Li ◽

Huiqun Jia

Keyword(s):

Gastric Cancer ◽

Cell Migration ◽

Cancer Cells ◽

Cell Lines ◽

Migration And Invasion ◽

Control Group ◽

Gastric Cancer Cells ◽

Cell Migration And Invasion ◽

Forkhead Box ◽

Induced Apoptosis

Objective Previous studies suggested that sevoflurane exerts anti-proliferative, anti-migratory, and anti-invasive effects on cancer cells. To determine the role of sevoflurane on gastric cancer (GC) progression, we evaluated its effects on the proliferation, migration, and invasion of SGC7901, AGS, and MGC803 GC cells. Methods GC cells were exposed to different concentrations of sevoflurane (1.7, 3.4, or 5.1% v/v). Cell viability, migration, and invasion were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Transwell assays. Immunohistochemical staining and immunoblotting were performed to analyze forkhead box protein 3 (FOXP3) protein expression in tissue specimens and cell lines, respectively. Results FOXP3 was downregulated in human GC specimens and cell lines. Functionally, FOXP3 overexpression significantly inhibited the proliferation, migration, and invasion of GC cells and accelerated their apoptosis. Moreover, sevoflurane significantly blocked GC cell migration and invasion compared with the findings in the control group. However, FOXP3 silencing neutralized sevoflurane-induced apoptosis and the inhibition of GC cell migration and invasion. Sevoflurane-induced apoptosis and the suppression of migration and invasion might be associated with FOXP3 overactivation in GC cells. Conclusions Sevoflurane activated FOXP3 and prevented GC progression via inhibiting cell migration and invasion in vitro.

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Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines

BMC Veterinary Research ◽

10.1186/s12917-020-02709-5 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Michela Levi ◽

Roberta Salaroli ◽

Federico Parenti ◽

Raffaella De Maria ◽

Augusta Zannoni ◽

...

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Cell Lines ◽

Tumour Cell ◽

S Phase ◽

Mammary Tumour ◽

Cancer Resistance ◽

Neoplastic Cells ◽

Canine Mammary Tumour ◽

Tumour Cell Lines

Abstract Background Doxorubicin (DOX) is widely used in both human and veterinary oncology although the onset of multidrug resistance (MDR) in neoplastic cells often leads to chemotherapy failure. Better understanding of the cellular mechanisms that circumvent chemotherapy efficacy is paramount. The aim of this study was to investigate the response of two canine mammary tumour cell lines, CIPp from a primary tumour and CIPm, from its lymph node metastasis, to exposure to EC50(20h) DOX at 12, 24 and 48 h of treatment. We assessed the uptake and subcellular distribution of DOX, the expression and function of P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP), two important MDR mediators. To better understand this phenomenon the effects of DOX on the cell cycle and Ki67 cell proliferation index and the expression of p53 and telomerase reverse transcriptase (TERT) were also evaluated by immunocytochemistry (ICC). Results Both cell lines were able to uptake DOX within the nucleus at 3 h treatment while at 48 h DOX was absent from the intracellular compartment (assessed by fluorescence microscope) in all the surviving cells. CIPm, originated from the metastatic tumour, were more efficient in extruding P-gp substrates. By ICC and qRT-PCR an overall increase in both P-gp and BCRP were observed at 48 h of EC50(20h) DOX treatment in both cell lines and were associated with a striking increase in the percentage of p53 and TERT expressing cells by ICC. The cell proliferation fraction was decreased at 48 h in both cell lines and cell cycle analysis showed a DOX-induced arrest in the S phase for CIPp, while CIPm had an increase in cellular death without arrest. Both cells lines were therefore composed by a fraction of cells sensible to DOX that underwent apoptosis/necrosis. Conclusions DOX administration results in interlinked modifications in the cellular population including a substantial effect on the cell cycle, in particular arrest in the S phase for CIPp and the selection of a subpopulation of neoplastic cells bearing MDR phenotype characterized by P-gp and BCRP expression, TERT activation, p53 accumulation and decrease in the proliferating fraction. Important information is given for understanding the dynamic and mechanisms of the onset of drug resistance in a neoplastic cell population.

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