Signaling crosstalk of FHIT, CHK2 and p38 in etoposide induced growth inhibition in MCF-7 cells

Mohammadrezaei, Fereshteh Mir; Kouchesfahani, Homa Mohseni; Montazeri, Hamed; Gharghabi, Mehdi; Ostad, Seyed Nasser; Ghahremani, Mohammad Hossein

doi:10.1016/j.cellsig.2012.09.019

Cited by 7 publications

(16 citation statements)

References 29 publications

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“…The siRNA oligo sequences (FHIT) were synthesized (Qiagen, Hilden, Germany) and cells were transfected with different amount of siRNA (10,20, and 30 nmol/L) into subconfluent cells using Lipofectamin 2000 reagent according to the manufacturer's instructions. After 24 hours, the cells were lysed, and the amount of FHIT protein was measured by Western blot analysis.…”

Section: Sirna Transfectionmentioning

confidence: 99%

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Signaling Crosstalk of FHIT, p53, and p38 in etoposide‐induced apoptosis in MCF‐7 cells

Khedri

Khaghani

Kheirollah

et al. 2019

J of Cellular Biochemistry

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Fragile histidine trail (FHIT) is a tumor suppressor in response to DNA damage which has been deleted in various tumors. However, the signaling mechanisms and interactions of FHIT with regard to apoptotic proteins including p53 and p38 in the DNA damage-induced apoptosis are not well described. In the present study, we used etoposide-induced DNA damage in MCF-7 as a model to address these crosstalks. The time course study showed that the expression of FHIT, p53, and p38MAPK started after 1 hour following etoposide treatment. FHIT overexpression led to increase p53 expression, p38 activation, and augmented apoptosis following etoposide-induced DNA damage compared to wild-type cells. However, FHIT knockdown blocked p53 expression, delayed p38 activation, and completely inhibited etoposide-induced apoptosis. Inhibition of p38 activity prevented induction of p53, FHIT, and apoptosis in this model. Thus, activation of p38 upon etoposide treatment leads to increase in FHIT and p53 expression. In p53 knockdown MCF-7, the FHIT induction was hampered but p38 activation was induced in lower doses of etoposide. In p53 knockdown cells, inhibition of p38 induced FHIT expression and apoptosis. Our data demonstrated that the exposure of MCF-7 cells to etoposide increases apoptosis through a mechanism involving the activation of the p38-FHIT-p53 pathway. Moreover, our findings suggest signaling interaction for these pathways may represent a promising therapy for breast cancer. K E Y W O R D S DNA damage, etoposide, FHIT, p38, p53 J Cell Biochem. 2019;120:9125-9137.wileyonlinelibrary.com/journal/jcb

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Section: Sirna Transfectionmentioning

confidence: 99%

“…28 Our group has previously shown that in response to etoposide, the FHIT expression is increased and p38 is activated. 20 However, the interplay of these proteins has not been defined. Thus, we have investigated the crosstalk of FHIT, p53, and p38MAPK in response to the etoposide-induced DNA damage perhaps to reveal some novel targets in cancer treatment.…”

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confidence: 99%

Signaling Crosstalk of FHIT, p53, and p38 in etoposide‐induced apoptosis in MCF‐7 cells

Khedri

Khaghani

Kheirollah

et al. 2019

J of Cellular Biochemistry

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“…Furthermore, peripheral blood samples show that active smokers express fragility at the FHIT locus. 13,[43][44][45] Relying on these data, we hypothesized that FHIT plays a role in mediating the cellular response to cigarette smoke. To test this, siRNA-treated HBEC3-TK cells were exposed to 1% CSE for 4 hours prior to RNA extraction and microarray analysis.…”

Section: Gene Expression Changes In Response To Fhit Knockdownmentioning

confidence: 99%

“…[43][44][45]57 Notably, modulation of these pathways has also been demonstrated to affect HMOX1 expression. [51][52][53][58][59][60] For this reason, we asked whether the superinduction of HMOX1 seen after FHIT knockdown might be mediated through one or more of these pathways.…”

Section: Mechanism Of Hmox1 Superinduction In Response To Fhit Lossmentioning

confidence: 99%

A knockdown with smoke model reveals FHIT as a repressor of Heme oxygenase 1

Boylston

Brenner²

2014

Cell Cycle

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Fragile histidine triad (FHIT) gene deletions are among the earliest and most frequent events in carcinogenesis, particularly in carcinogen-exposed tissues. Though FHIT has been established as an authentic tumor suppressor, the mechanism underlying tumor suppression remains opaque. Most experiments designed to clarify FHIT function have analyzed the consequence of re-expressing FHIT in FHIT-negative cells. However, carcinogenesis occurs in cells that transition from FHIT-positive to FHIT-negative. To better understand cancer development, we induced FHIT loss in human bronchial epithelial cells with RNA interference. Because FHIT is a demonstrated target of carcinogens in cigarette smoke, we combined FHIT silencing with cigarette smoke extract (CSE) exposure and measured gene expression consequences by RNA microarray. The data indicate that FHIT loss enhances the expression of a set of oxidative stress response genes after exposure to CSE, including the cytoprotective enzyme heme oxygenase 1 (HMOX1) at the RNA and protein levels. Data are consistent with a mechanism in which Fhit protein is required for accumulation of the transcriptional repressor of HMOX1, Bach1 protein. We posit that by allowing superinduction of oxidative stress response genes, loss of FHIT creates a survival advantage that promotes carcinogenesis.

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“…Ideally, we can evaluate a proposed experimental protocol (using a specific set of instruments and analysis software) by comparing its measurements for known cell lines against accepted "gold standard" values as ground truth. However, considerable variability in phenotypic measurements has been observed even for widely used cell lines (e.g., see the discordant cell population doubling times for MCF-7 breast cancer lines grown in "standard" normoxic conditions in [9][10][11][12][13][14][15][16][17].) Thus, it is difficult to assess the impact of experimental design choices and evaluate the quality of new measurements without well-established ground truths for experimental measurements.…”

Section: Introductionmentioning

confidence: 99%

Computational investigation of biological and technical variability in high throughput phenotyping and cell line identification

Friedman

Macklin

2017

Preprint

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Abstract:High-throughput cell profiling experiments are characterizing cell phenotype under a broad variety of microenvironmental and therapeutic conditions. However, biological and technical variability are contributing to wide ranges of reported parameter values, even for standard cell lines grown in identical conditions. In this paper, we develop a mathematical model of cell proliferation assays that account for biological and technical variability and limitations of the experimental platforms, including (1) cell confluency effects, (2) biological variability and technical errors in pipetting, (3) biological variability in proliferation characteristics, (4) technical variability and uncertainty in measurement timing, (5) cell counting errors, and (6) the impact of limited temporal sampling. We use this model to create synthetic datasets with growth rates and measurement times typical of cancer cell cultures, and investigate the impact of the initial cell seeding density and the common practice of fitting exponential growth curves to three cell count measurements. We find that the combined sources of variability mask the sub-exponential growth characteristics of the synthetic datasets, and that researchers profiling the same cell lines under different seeding characteristics can find significant (p < 0.05) differences in the measured growth rates. Even seeding the cells at 1% of the confluent limit can cause significant (p < 0.05) differences in the measured growth rate from the ground truth. We explored the effect of reducing errors in each part of the virtual experimental system, and found the best improvements from reducing timing errors, reducing cell counting errors, or reducing the interval between measurements (to reduce the inaccuracy of the exponential growth assumption when fitting curves). Reducing biological variability and pipetting errors had the least impact, because any improvements are still masked by cell counting errors. We close with a discussion of recommended practices for high-throughput cell phenotyping and cell line identification systems.

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Signaling crosstalk of FHIT, CHK2 and p38 in etoposide induced growth inhibition in MCF-7 cells

Cited by 7 publications

References 29 publications

Signaling Crosstalk of FHIT, p53, and p38 in etoposide‐induced apoptosis in MCF‐7 cells

Signaling Crosstalk of FHIT, p53, and p38 in etoposide‐induced apoptosis in MCF‐7 cells

A knockdown with smoke model reveals FHIT as a repressor of Heme oxygenase 1

Computational investigation of biological and technical variability in high throughput phenotyping and cell line identification

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