Expression of TCF3 in Wilms' tumor and its regulatory role in kidney tumor cell viability, migration and apoptosis in vitro

Purpose: Compelling evidence has unveiled the importance of long noncoding RNAs (lncRNAs) in malignant behavior of Wilms’ tumor (WT). Hereon, we intend to assess the function and associated molecular mechanism of lncRNA maternally expressed gene 8 (MEG8) in WT cells. Methods: Expression levels of MEG8, miR-23a-3p, and CT10 regulator of kinase (CRK) were determined by quantitative real-time polymerase chain reaction. Cell viability was assessed by MTT assay. Besides, wound healing assay and transwell assay were applied to examine abilities of cell migration and invasion, respectively. Dual-luciferase reporter assay was employed to test the interplay among MEG8, miR-23a-3p, and CRK. Western blot was used to detect relative protein expression of CRK. Results: MEG8 and CRK expression was elevated, while miR-23a-3p expression was decreased in WT tissues and cells. The histologic type, lymphatic metastasis, and National Wilms Tumor Study (NWTS) stage were associated with the expression of MEG8, miR-23a-3p, and CRK in WT patients. MEG8 knockdown or miR-23a-3p overexpression restrained WT cells in cell viability, migration, and invasiveness in vitro. As to mechanism exploration, MEG8 could directly bind to miR-23a-3p and then miR-23a-3p targeted CRK. MEG8 was inversely correlated with miR-23a-3p and positively correlated with CRK in WT tissues. Meantime, miR-23a-3p was inversely correlated with CRK in WT tissues. Additionally, MEG8 knockdown-mediated suppressive impacts on cell viability, migration, and invasiveness were reversed by overexpression of CRK or repression of miR-23a-3p in WT cells. Conclusions: The cell viability, migration, and invasiveness of WT cells were repressed by MEG8 knockdown via targeting the miR-23a-3p/CRK axis.

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In Vitro Effect of Focused Ultrasound or Thermal Stress on HSP70 Expression and Cell Viability in Three Tumor Cell Lines

Academic Radiology ◽

10.1016/j.acra.2007.04.008 ◽

2007 ◽

Vol 14 (7) ◽

pp. 859-870 ◽

Cited By ~ 27

Author(s):

Walter Hundt ◽

Caitlin E. O’Connell-Rodwell ◽

Mark D. Bednarski ◽

Silke Steinbach ◽

Samira Guccione

Keyword(s):

Thermal Stress ◽

Tumor Cell ◽

Cell Viability ◽

Cell Lines ◽

Focused Ultrasound ◽

Hsp70 Expression ◽

Tumor Cell Lines ◽

Vitro Effect

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Aptamer-polymer functionalized silicon nanosubstrates for enhanced recovered circulating tumor cell viability and in vitro chemosensitivity testing

International Journal of Nanomedicine ◽

10.2147/ijn.s103569 ◽

2016 ◽

pp. 2133 ◽

Cited By ~ 3

Author(s):

Qinglin Shen ◽

Caixia Peng ◽

Yan Zhan ◽

Liang Fan ◽

Mengyi Wang ◽

...

Keyword(s):

Tumor Cell ◽

Cell Viability ◽

Circulating Tumor Cell ◽

Chemosensitivity Testing ◽

In Vitro Chemosensitivity

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Photodynamic inactivation of selected intracellular enzymes by hematoporphyrin derivative and their relationship to tumor cell viability in vitro

Cancer Letters ◽

10.1016/0304-3835(84)90132-0 ◽

1984 ◽

Vol 24 (2) ◽

pp. 165-172 ◽

Cited By ~ 45

Author(s):

Russell Hilf ◽

Nicholas W. Warne ◽

Douglas B. Smail ◽

Scott L. Gibson

Keyword(s):

Tumor Cell ◽

Cell Viability ◽

Photodynamic Inactivation ◽

Hematoporphyrin Derivative ◽

Intracellular Enzymes

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Therapeutic Potential of Pharmacological Targeting NLRP3 Inflammasome Complex in Cancer

Frontiers in Immunology ◽

10.3389/fimmu.2020.607881 ◽

2021 ◽

Vol 11 ◽

Author(s):

Gulcin Tezcan ◽

Ekaterina E. Garanina ◽

Mohammad Alsaadi ◽

Zarema E. Gilazieva ◽

Ekaterina V. Martinova ◽

...

Keyword(s):

Complex Formation ◽

Tumor Cell ◽

Cell Viability ◽

Nlrp3 Inflammasome ◽

Therapeutic Potential ◽

Human Fibroblasts ◽

A549 Cells ◽

Annexin V ◽

Tube Formation

IntroductionDysregulation of NLRP3 inflammasome complex formation can promote chronic inflammation by increased release of IL-1β. However, the effect of NLRP3 complex formation on tumor progression remains controversial. Therefore, we sought to determine the effect of NLRP3 modulation on the growth of the different types of cancer cells, derived from lung, breast, and prostate cancers as well as neuroblastoma and glioblastoma in-vitro.MethodThe effect of Caspase 1 inhibitor (VX765) and combination of LPS/Nigericin on NLRP3 inflammasome activity was analyzed in A549 (lung cancer), MCF-7 (breast cancer), PC3 (prostate cancer), SH-SY5Y (neuroblastoma), and U138MG (glioblastoma) cells. Human fibroblasts were used as control cells. The effect of VX765 and LPS/Nigericin on NLRP3 expression was analyzed using western blot, while IL-1β and IL-18 secretion was detected by ELISA. Tumor cell viability and progression were determined using Annexin V, cell proliferation assay, LDH assay, sphere formation assay, transmission electron microscopy, and a multiplex cytokine assay. Also, angiogenesis was investigated by a tube formation assay. VEGF and MMPs secretion were detected by ELISA and a multiplex assay, respectively. Statistical analysis was done using one-way ANOVA with Tukey’s analyses and Kruskal–Wallis one-way analysis of variance.ResultsLPS/Nigericin increased NRLP3 protein expression as well as IL-1β and IL-18 secretion in PC3 and U138MG cells compared to A549, MCF7, SH-SY5Y cells, and fibroblasts. In contrast, MIF expression was commonly found upregulated in A549, PC3, SH-SY5Y, and U138MG cells and fibroblasts after Nigericin treatment. Nigericin and a combination of LPS/Nigericin decreased the cell viability and proliferation. Also, LPS/Nigericin significantly increased tumorsphere size in PC3 and U138MG cells. In contrast, the sphere size was reduced in MCF7 and SH-SY5Y cells treated with LPS/Nigericin, while no effect was detected in A549 cells. VX765 increased secretion of CCL24 in A549, MCF7, PC3, and fibroblasts as well as CCL11 and CCL26 in SH-SY5Y cells. Also, VX765 significantly increased the production of VEGF and MMPs and stimulated angiogenesis in all tumor cell lines.DiscussionOur data suggest that NLRP3 activation using Nigericin could be a novel therapeutic approach to control the growth of tumors producing a low level of IL-1β and IL-18.

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Using a hybrid radioenhancer to realize tumor cell-targeted treatment for osteosarcoma: an in vitro study

Current Medicinal Chemistry ◽

10.2174/0929867327666201118155216 ◽

2020 ◽

Vol 27 ◽

Author(s):

Fu-I Tung ◽

Li-Chin Chen ◽

Yu-Chi Wang ◽

Ming-Hong Chen ◽

Pei-Wei Shueng ◽

...

Keyword(s):

Western Blot ◽

Tumor Cell ◽

Cell Viability ◽

Tumor Cells ◽

Clonogenic Assay ◽

Normal Cells ◽

Targeted Radiotherapy ◽

In Vitro Investigation ◽

Dose Radiation

: Osteosarcoma is insensitive to radiation. High-dose radiation is often used as a treatment, but causes side effects in patients. Hence, it is important to develop tumor cell-targeted radiotherapy that could improve radiotherapy efficiency on tumor cells and reduce the toxic effect on normal cells during radiation treatment. In this study, we developed an innovative method for treating osteosarcoma by using a novel radiation-enhancer (i.e., carboxymethyl-hexanoyl chitosan-coated selfassembled Au@Fe3O4 nanoparticles; CSAF NPs). CSAF NPs were employed together with 5-aminolevulinic acid (5-ALA) to achieve tumor cell-targeted radiotherapy. In this study, osteosarcoma cells (MG63) and normal cells (MC3T3-E1) were used for an in vitro investigation, in which a reactive oxygen species (ROS) assay, cell viability assay, clonogenic assay, and western blot were used to confirm the treatment efficiency. The ROS assay showed that the combination of CSAF NPs and 5-ALA enhanced radiation-induced ROS production in tumor cells (MG63); however, this was not observed in normal cells (MC3T3-E1). The cell viability ratio of normal cells to tumor cells after treatment with CSAF NPs and 5-ALA reached 2.79. Moreover, the clonogenic assay showed that the radiosensitivity of MG63 cells was increased by the combination use of CSAF NPs and 5-ALA. This was supported by performing a western blot that confirmed expression of cytochrome c (a marker of cell mitochondria damage) and caspase-3 (a marker of cell apoptosis). The results provide an essential basis for developing tumor-cell targeted radiotherapy by means of low-dose radiation.

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Essential experimental steps and estimates of renal carcinoma initiating cells.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.15_suppl.11127 ◽

2014 ◽

Vol 32 (15_suppl) ◽

pp. 11127-11127

Author(s):

Craig Gedye ◽

Danylo Sirskyj ◽

Nazleen Carol Lobo ◽

Ella Hyatt ◽

Andrew Evans ◽

...

Keyword(s):

Tumor Cell ◽

Cell Viability ◽

Tumor Cells ◽

Cell Lines ◽

Stromal Cells ◽

Ex Vivo ◽

Rock Inhibitor ◽

Cell Renal Cell Carcinoma ◽

Cell Potential

11127 Background: Rare cancer stem cells (CSC), proposed to be solely responsible for tumor propagation and re-initiation, are functionally identified as tumor-initiating cells (TIC) from ex vivo tumors using xenotransplantation and clonogenic limiting dilution assays (LDA). TIC have not previously been described from ex vivohuman clear cell renal cell carcinoma (ccRCC). Methods: Primary human ccRCC samples (n=120) from patients undergoing nephrectomy were processed and implanted as subcapsular fragments or cell suspension injection LDAs with Matrigel in NOD/SCID/IL2Rγ-/- (NSG) mice, and observed for at least 6 months. In vitro clonogenic LDAs assays were performed from primary cell suspensions and ccRCC cell lines. LDAs were supplemented with human stromal cells and proteins, and the Y-26732 ROCK inhibitor. Multiparametric flow cytometry and immunofluorescence were used to investigate tumor heterogeneity and cell viability. Results: ccRCC TIC appeared rare from injected suspensions, but xenografts engrafted frequently from tiny fragments, and clonogenic frequencies were 103-104greater than TIC frequencies, suggesting that LDAs underestimated ccRCC tumor cell potential. We systematically identified multiple methodological steps that distort quantitation and identification of ccRCC TIC. For example cell viability was highly variable prior to processing, disaggregation itself destroyed up to 99% of tumor cells, standard assays substantially overestimated tumor cell viability in suspensions, and supplementation with human extracellular cells or proteins, or inhibition of anoikis by Y-26732 increased clonogenic and TIC frequencies in cell lines and primary ccRCC suspensions. Annexin-V staining revealed that tumor cells were more apoptotic then normal stromal cells, and that tumor cells positive for CD44 (a putative CSC marker) were more viable than CD44- tumor cells. Conclusions: We describe multiple, unappreciated and largely unavoidable observational errors in essential methods used to study TIC in ccRCC. ccRCC TIC may be more common than appreciated. Re-examination of the CSC hypothesis in other solid tumors is warranted in view of these previously unexplored methodological biases.

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