A single nuclear polymorphism in let‐7g binding site affects the doubling time of thyroid nodule by regulating KRAS‐induced cell proliferation

2019 ◽  
Vol 234 (12) ◽  
pp. 23437-23447
Author(s):  
Ailin Liu ◽  
Wanli Zhang ◽  
Tao Zhao ◽  
Ming Xiao ◽  
Qijian Mei ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Thyroid Nodule ◽  
Binding Site ◽  
Doubling Time ◽  
Nuclear Polymorphism ◽  
Single Nuclear Polymorphism

THYROID NODULE GROWTH AS A PREDICTOR OF MALIGNANCY

2019 ◽  
Vol 25 (10) ◽  
pp. 1029-1034 ◽  
Author(s):  
Kathleen O'Connell ◽  
Alexa Clark ◽  
Wilma Hopman ◽  
Joshua Lakoff
Keyword(s):  
Confidence Interval ◽  
Thyroid Nodule ◽  
Odds Ratio ◽  
Doubling Time ◽  
Thyroid Nodules ◽  
Volume Growth ◽  
Needle Aspiration ◽  
Health Science ◽  
Significant Difference ◽  
Nodule Growth

Objective: To assess which measure of thyroid nodule growth on serial neck ultrasound, if any, is associated with malignancy. Methods: Retrospective exploratory chart review of malignant thyroid nodules assessed at Kingston Health Sciences Centre (2006–2016) and benign thyroid nodules (2016), at least 1 cm in diameter and with 2 ultrasounds completed at least 30 days apart. Groups were compared using independent samples Student's t test, chi-square test, or Mann-Whitney U test as appropriate, as well as multivariable logistic and linear regression modelling to adjust for age and baseline volume. Results: One hundred and seventy-eight nodules were included in the study. When growth was defined as >20% increase in 2 dimensions (minimum 2 mm), malignant nodules (MNs) underwent significantly more growth than benign nodules (BNs) (16.8% BN versus 29.8% MN [ P = .026]; odds ratio = 2.49; 95% confidence interval = 1.12 to 5.56). There was no significant difference between the groups when growth was defined as >2 mm/year or ≥50% volume growth. Nodules shrank >2 mm/year in each group and the difference was not statistically significant (24.2% BN versus 20.7% MN [ P = .449]). The median doubling time for the nodules that grew was 1022.1 days in the BN group and 463.2 days in the MN group ( P = .036). The median doubling time for all nodules was 456.5 days in the BN group and 244.2 days in the MN group ( P = .015). Conclusion: Thyroid nodule growth defined as >20% increase in 2 dimensions (minimum 2 mm) is associated with risk of malignancy. Nodule shrinkage did not distinguish between BNs and MNs. Abbreviations: BN = benign nodule; CI = confidence interval; FNA = fine needle aspiration; KHSC = Kingston Health Science Centre; MN = malignant nodule; OR = odds ratio; ROC = receiver operating characteristic

Chordoma of the skull base: predictors of tumor recurrence

2003 ◽  
Vol 98 (4) ◽  
pp. 812-822 ◽  
Author(s):  
Roberto Pallini ◽  
Giulio Maira ◽  
Francesco Pierconti ◽  
Maria Laura Falchetti ◽  
Ester Alvino ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Microsatellite Instability ◽  
Skull Base ◽  
Tumor Recurrence ◽  
Doubling Time ◽  
P53 Protein ◽  
Biological Behavior ◽  
Htert Mrna ◽  
Content Type ◽  
Fine Print

Object. Chordomas of the skull base are generally regarded as slow-growing tumors; however, approximately 20% of these lesions have been shown to recur as early as 1 year postsurgery. The classic pathological paradigms are poor predictors of outcome, and additional markers are needed to identify patients at risk for early tumor recurrence. In this study the authors describe such a marker. Methods. In a series of 26 patients with chordomas of the skull base, the authors investigated the relationship between the biological behavior of the tumor, which was determined according to the interval for its recurrence and volume doubling time, and several pathological and molecular features, which included the histological variant, proliferative activity, mutation of p53 protein, expression of human telomerase reverse transcriptase (hTERT) messenger (m)RNA, loss of heterozygosity (LOH), and microsatellite instability. The major finding in this study was that hTERT mRNA expression in chordoma cells identifies those tumors that exhibit unusually fast rates of growth. The expression of hTERT mRNA was frequently associated with mutation of p53 protein, indicating that telomerase dysfunction combines with abnormal p53 function to initiate the unrestrained clonal expansion of the tumor cells. In cases in which the tumor was partially removed, mutation of p53 protein and expression of hTERT mRNA predicted increased doubling time for residual tumor as well as the probability of tumor recurrence. Cell proliferation, as investigated using the Ki-67 method, was significantly related to the tumor doubling time; however, the authors found that the pattern of cell proliferation was not homogeneous throughout the chordoma tissue, and that the proliferative index might change by a factor as high as 8 among different regions of the same tumor. The LOH and microsatellite instability do not seem to affect the prognosis of skull base chordomas. Conclusions. Reactivation of telomerase in chordomas is a reliable predictor of outcome. The ability to predict the biological behavior of chordomas might have immediate implications in the management of this disease in patients who undergo surgery.

scholarly journals miR-192-5p suppresses the progression of lung cancer bone metastasis by targeting TRIM44

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Peng Zou ◽  
Menghai Zhu ◽  
Chong Lian ◽  
Jiaqiang Wang ◽  
Zhiquan Chen ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Bone Metastasis ◽  
Binding Site ◽  
Cancer Cells ◽  
Significant Role ◽  
Research Group ◽  
Luciferase Activity ◽  
Migration And Invasion

AbstractLung cancer is the leading cause of cancer-related deaths worldwide, with 50–70% of patients suffering from bone metastasis. Accumulating evidence has demonstrated that miRNAs are involved in cell proliferation, migration, and invasion in malignancy, such as lung cancer bone metastasis. In the present study, we demonstrated that reduced miR-192-5p and increased TRIM44 levels were associated with the proliferation, migration and invasion of lung cancer. Furthermore, the potential functions of miR-192-5p were explored in A549 and NCI-H1299 cells. We found that miR-192-5p upregulation suppressed tumour behaviours in lung cancer cells. To further investigate whether miR-192-5p is associated with TRIM44, we used TargetScan software to predict the binding site between miR-192-5p and TRIM44. Luciferase activity assays were performed to verify this prediction. In addition, the significant role of miR-192-5p in negatively regulating TRIM44 expression was manifested by our research group. our results suggest that miR-192-5p inhibited the proliferation, migration and invasion of lung cancer through TRIM44.

The expression of the tumour suppressor HBP1 is down-regulated by growth factors via the PI3K/PKB/FOXO pathway

2014 ◽  
Vol 460 (1) ◽  
pp. 25-36 ◽  
Author(s):  
Alexandra Coomans de Brachène ◽  
Emeline Bollaert ◽  
Astrid Eijkelenboom ◽  
Audrey de Rocca Serra ◽  
Kristan E. van der Vos ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Growth Factors ◽  
Binding Site ◽  
Tumour Suppressor

We identified HBP1 as a gene that is down-regulated by growth factors through the PI3K/PKB/FOXO pathway. A FOXO-binding site was characterized in the HBP1 promoter. HBP1 knockdown was enough to enhance cell proliferation downstream of growth factors.

scholarly journals Comparison of BrdUrd and [3H]TdR incorporation to estimate cell proliferation, cell loss, and potential doubling time in tumor xenografts

Cytometry ◽  
1992 ◽  
Vol 13 (8) ◽  
pp. 872-879 ◽  
Author(s):  
Ulf K. Zätterström ◽  
Maria Johansson ◽  
Anders Källén ◽  
Bo Baldetorp ◽  
Stina Oredsson ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Doubling Time ◽  
Cell Loss ◽  
Tumor Xenografts ◽  
Potential Doubling Time ◽  
Potential Doubling

scholarly journals Blocking Lysosomal Two-Pore Channel 2 Function Inhibits Proliferation of Multidrug Resistant Leukemia Cells and Sensitizes Them to Vincristine Treatment

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 2081-2081
Author(s):  
Martin Müller ◽  
Franz Geisslinger ◽  
Susanne Gerndt ◽  
Ramona Schütz ◽  
Yu-Kai Chao ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Patch Clamp ◽  
Mrna Expression ◽  
Treatment Response ◽  
Cell Line ◽  
Doubling Time ◽  
Multidrug Resistant ◽  
Pore Channel ◽  
Increased Sensitivity

Introduction Two-pore channel 2 (TPC2) represents an exceptional ion channel due to its localization on lysosomes1. TPC2 is overexpressed in several cancer types, e.g. in prostate cancer, and its expression is associated with poor survival probability2. To date, a role of TPC2 in leukemia has not been investigated. Pharmacologic inhibition of TPC2 can be performed with tetrandrine (Tet), a bisbenzylisoquinoline alkaloid3. However, its use is restricted due to unfavorable toxicity in animal studies4. In our study, we aim to elucidate the role of TPC2 function in multidrug resistant leukemia and, concurrently, identify efficacious TPC2 inhibitors by screening a library of synthetic bisbenzylisoquinoline derivatives (BBIQDs). Results Firstly, via qPCR analysis, we detected a 2-fold increased TPC2 mRNA expression in a vincristine-resistant (VCR-R) CEM (VCR-R CEM, T-ALL) cell line, compared with the parental CCRF-CEM cell line. Secondly, we knocked out TPC2 in VCR-R CEM cells using the CRISPR-Cas9 system to investigate its involvement in cell growth and treatment response. By studying cell proliferation over a period of six days, we found that the doubling time of TPC2-deficient cells was increased to 30 h, compared with 24 h for the wildtype (wt) cell line. Additionally, treatment of TPC2 knockout (ko) cells with VCR resulted in increased sensitivity versus wt, as indicated by proliferation (IC50 wt: 3.3 µM, IC50 ko: 1.6 µM, 72 h) and apoptosis (EC50 wt: 3.0 µM, EC50 ko: 1.7 µM, 48 h) assays. Next, a library of BBIQDs was synthesized and screened for the ability to inhibit TPC2 function using the whole endolysosomal patch clamp method. Our small molecule hits SG-005 and SG-094 inhibited TPC2 current density by 50% and 70%, respectively, giving novel TPC2 inhibitors with either similar or even increased potency, compared to Tet (50%). Treatment of VCR-R CEM cells with Tet, SG-005 and 94 effectively inhibited proliferation (IC50: 5-15 µM, 48 h). Toxicity was assessed by propidium iodide exclusion assays using Peripheral Blood Mononuclear cells (PBMCs, n=3 healthy donors), indicating reduced toxicity of SG-094 (<5% dead cells vs. 25% dead cells for Tet and SG-005, 20 µM, 48 h treatment). Additionally, SG-094 was well tolerated in vivo when applied to a mouse model (90 nmol/kg/d on three consecutive days). Remarkably, combination of VCR (0.01 and 0.1 µM) with Tet, SG-005 or 94 (1 and 5 µM) synergistically increased treatment response of VCR-R CEM cells (wt) and of B ALL patient-derived xenograft (PDX) cells from a relapse patient (Bliss values: 1.1-2.4). Methods CellTiter-Blue® cell proliferation assays, qPCR mRNA expression, propidium iodide exclusion and analysis of apoptosis were performed as described by the manufacturers. Whole endolysosomal patch clamp, analysis of specific apoptosis of PDX cells and generation of the TPC2 ko model using the CRISPR-Cas9 system were performed as described previously3, 5, 6. Analysis of doubling time was performed by counting cells daily using trypan blue staining and a Vi-CELL XR device as indicated by the manufacturer. Conclusion Taken together, we detected an upregulated mRNA expression of endolysosomal TPC2 in VCR-R CEM cells, indicating that it represents a potential target for therapeutic intervention for difficult-to-treat leukemia phenotypes. Furthermore, we could show that genetic ablation of the ion channel results in a reduced proliferation rate and an increased sensitivity towards VCR treatment. Finally, we successfully identified pharmacologic TPC2 inhibitors which show antiproliferative effects when applied as monotherapy and synergistically enhance treatment response of VCR-R CEM cells and PDX cells when combined with VCR. References S. Patel, B. S. Kilpatrick, Biochim Biophys Acta Mol Cell Res1865, 1678-1686 (2018). F. Li, J. P. Ji, Y. Xu, R. L. Liu, Clin Transl Oncol, (2019). O. N. Nguyen et al., Cancer Res77, 1427-1438 (2017). H. Jin et al., Chem Res Toxicol24, 2142-2152 (2011). F. Koczian et al., Haematologica104, 546-555 (2019). F. A. Ran et al., Nat Protoc8, 2281-2308 (2013). Disclosures No relevant conflicts of interest to declare.

scholarly journals Hyperglycemia and hyperinsulinemia stimulate cervical cancer cell proliferation in vitro and in vivo study

2019 ◽  
Author(s):  
Miriam Gutiérrez-Gutiérrez ◽  
Ishell Aline Figueroa-Martínez ◽  
Rafael Jurado ◽  
Norma Uribe ◽  
Patricia García-López ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Cell Proliferation ◽  
Cancer Cell ◽  
Doubling Time ◽  
Xenograft Model ◽  
Cervical Cancer Cell ◽  
Diabetic Patients ◽  
Cell Doubling Time

Abstract Background: Diabetes mellitus and malignant tumor are the second and third causes of women death in Mexico. Hyperglycemia, insulin and insulin-like growth factor 1 are the main risk factors involved in cancer development in patient with diabetes. The aim of this study was to evaluate the effect of hyperglycemia and hyperinsulinemia over cell proliferation and tumor growth in cervical cancer. Methods: Cell proliferation, apoptosis and cell cycle of cervical cancer cell lines (HeLa, SiHa and CaSki) in presence of hyperglycemia and/or insulin were evaluated. Xenograft model for cervical cancer was done in diabetic female nu/nu mice; biochemical parameters, body weight, tumoral volume and cell doubling time were evaluated. Results: Hyperglycemia and hyperinsulinemia significantly increase cell proliferation and decreases apoptosis with no change in cell cycle. Insulin treatment increase tumor volume and diminish cell doubling time, this group also developed hyperinsulinemia and in Langerhans pancreatic islet hypertrophy; whereas, hyperglycemic groups show the same effects but in lesser degree than the insulin treated group. Conclusion: Glucose and insulin stimulates both, proliferation and tumoral growth of cervical cancer, so this should be a possible explanation for the low survival of diabetic patients with cervical cancer in compare to non-diabetic patients with cervical cancer.

Abstract 16176: Newly Identified MiR-6770-3p Dramatically Induces Angiogenesis in Human Endothelial Cells While Uncommonly Decreasing Cell Proliferation

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Marie Besnier ◽  
Owen Tang ◽  
Elijah Genetzakis ◽  
Meghan Finemore ◽  
Belinda Di Bartolo ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Binding Site ◽  
Luciferase Activity ◽  
Umbilical Vein ◽  
Tube Formation ◽  
Luciferase Reporter ◽  
Mrna Levels ◽  
Mrna Targets ◽  
Micro Rnas

Micro-RNAs (miRs) are small non-coding RNAs that alter the expression of multiple mRNA targets. Although they participate in physiological processes, dysregulation of their expression is implicated in various diseases. We investigated whether miRs could be involved in the regulation of FXYD1 expression, a sub-unit of the Na+/K+-ATPase pump, that we previously described as both cardio- and vasculoprotective, under oxidative stress conditions. Using in silico analysis, we identified 3 potential miRs that could target FXYD1 mRNA: miR-3178, miR-3960 and miR-6770-3p. Using a FXYD1-3’UTR-luciferase reporter assay, we found that the overexpression of miR-6770-3p in HEK293T cells resulted in the largest reduction in luciferase activity, showing a strong targeting of FXYD1 mRNA. A mutation of the binding site in the 3’UTR of FXYD1 restored luciferase activity, confirming the binding site of miR-6770-3p. Mir-6770-3p is a novel miR that currently has no known function. MiR-6770-3p is the minor strand of its 5p/3p miR duplex in both the human endothelial cell (EC) line, EA.hy.926, and human dermal microvascular (HMVECs). Its overexpression dramatically increased endothelial tube formation in a Matrigel angiogenesis assay (>4 fold change vs. control transfection, p<0.05) and was also associated with a 2-3-fold increase of VEGFR2 and Endoglin mRNA levels, respectively, in both cell types. Conversely, it was also uncommonly associated with a decrease in cell proliferation, suggesting that the pro-angiogenic effect of miR-6770-3p was not due to increased proliferation. Interestingly and contrary to our previous findings, miR-6770-3p was the major strand in Human Umbilical Vein ECs (HUVECs). In this cell type, overexpression of the 3p strand in fact decreased tube formation, while retaining its negative effect on proliferation. The difference in miR major and minor strand may partly explain the differences in functional behaviours in HUVECs, as compared to HMVECs. In conclusion, we have identified a novel miR which has the potential to modulate vessel formation in vitro, and induce critical regulators of angiogenesis; however, further work is required to better understand how this miR performs these functions.

scholarly journals In Vitro and in Silico Evaluation of Bikaverin as a Potent Inhibitor of Human Protein Kinase CK2

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1380 ◽  
Author(s):  
Samer Haidar ◽  
Dagmar Aichele ◽  
Robin Birus ◽  
Janine Hielscher ◽  
Tuomo Laitinen ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Protein Kinase ◽  
Cell Viability ◽  
Binding Site ◽  
Protein Kinase Ck2 ◽  
Potent Inhibitor ◽  
Cell Permeability ◽  
Atp Binding Site ◽  
Kinase Ck2

Protein kinase CK2 is an emerging target for therapeutic intervention in human diseases, particularly in cancer. Inhibitors of this enzyme are currently in clinical trials, indicating the druggability of human CK2. By virtual screening of the ZINC database, we found that the natural compound bikaverin can fit well in the ATP binding site of the target enzyme CK2. By further in vitro evaluation using CK2 holoenzyme, bikaverin turned to be a potent inhibitor with an IC50 value of 1.24 µM. In this work, the cell permeability of bikaverin was determined using a Caco-2 cell permeability assay as a prerequisite for cellular evaluation and the compound turned out to be cell permeable with a Papp- value of 4.46 × 10−6 cm/s. Bikaverin was tested for its effect on cell viability using a MTT assay and cell proliferation using an EdU assay in different cancer cell lines (MCF7, A427 and A431 cells). Cell viability and cell proliferation were reduced dramatically after treatment with 10 µM bikaverin for 24 h. Additionally the IncuCyte® live-cell imaging system was applied for monitoring the cytotoxicity of bikaverin in the three tested cancer cell lines. Finally, molecular dynamic studies were performed to clarify the ligand binding mode of bikaverin at the ATP binding site of CK2 and to identify the amino acids involved.

Sign in / Sign up

Export Citation Format

Share Document