Functional specific roles of <scp>H</scp>‐<i>ras</i> and <scp>N</scp>‐<i>ras</i>. A proteomic approach using knockout cell lines

Ferreira, Laura; Fuentes-Calvo, Isabel; Muñoz‐Félix, José M.; Muñiz-Martín, Carmen; Sánchez-Juanes, Fernando; Raposo, César; González-Buitrago, José Manuel; López‐Novoa, José M.; Martı́nez-Salgado, Carlos

doi:10.1002/elps.201100606

Cited by 4 publications

(3 citation statements)

References 63 publications

(70 reference statements)

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“…How KRAS mutation drives ERCC1 downregulation remains to be understood, but recently it has been shown that in breast cancer cells the expression of ERCC1 is regulated by H-RAS, one of the three proteins of the RAS family, through AP-1 29 . Contrary to what was observed in the past, it has also been shown that the three protein of RAS family (N-RAS, KRAS and H-RAS), have different activities and each may affect the synthesis and function of the others 30 - 31 . Therefore it is conceivable that mutational status of KRAS can influence the activity of H-RAS, and consequently the expression of ERCC1.…”

Section: Discussioncontrasting

confidence: 57%

ERCC1 Induction after Oxaliplatin Exposure May Depend on KRAS Mutational Status in Colorectal Cancer Cell Line: In Vitro Veritas

Orlandi¹,

Salvatore²,

Bagalá³

et al. 2015

J. Cancer

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Introduction: Oxaliplatin (Oxa) is widely used in metastatic colorectal cancer (mCRC), but currently there are not valid predictors of response to this drug. In the control arms both of OPUS and PRIME studies Oxa seems more active in patients with mCRC with mutated (mt) KRAS than in those with wild type (wt) KRAS. Recently we have retrospectively confirmed this suggestion, therefore we have hypothesized that the mutational status of KRAS could influence the expression of ERCC1, one of the main mechanisms of Oxa resistance.Material and Methods: We used four cell lines of colorectal cancer: two KRAS wild type (wt) (HCT-8 and HT-29) and two KRAS mt (SW620 and SW480). We evaluated the sensitivity of these cell lines to Oxa by MTT-test as well the ERCC1 levels before and after 24 h exposure to Oxa by Real-Time PCR. We silenced KRAS in a KRAS mt cell line (SW620LV) to evaluate the impact on Oxa sensitivity and ERCC1 levels. Lastly, ERCC1 was also silenced in order to confirm the importance of this protein as an Oxa resistance factor.Results: The KRAS mt cell lines resulted more sensitive to Oxa (OR 2.68; IC 95% 1.511-4.757 p<0.001). The basal levels of ERCC1 did not show significant differences between KRAS mt and wt cell lines, however, after 24 h exposure to Oxa, only the wt KRAS lines showed the ability to induce ERCC1, with a statistically significant difference (OR 42.9 IC 95% 17.260-106.972 p<0.0005). By silencing KRAS, sensitivity to Oxa was reduced in mt KRAS cell lines and this effect was associated with the acquisition of ability to induce ERCC1. Silencing of ERCC1, in turn, enhanced the sensitivity to Oxa in wt KRAS cell lines and restored sensitivity to Oxa in SW620LV cell line.Conclusion: KRAS mutated cell lines were more sensitive to Oxa. This feature seems secondary to the inability of these cells to induce ERCC1 after exposure to Oxa. Thus, KRAS mutational status might be a predictor of response to Oxa in CRC surrogating the cell ability to induce ERCC1.

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Section: Discussioncontrasting

confidence: 57%

ERCC1 Induction after Oxaliplatin Exposure May Depend on KRAS Mutational Status in Colorectal Cancer Cell Line: In Vitro Veritas

Orlandi¹,

Salvatore²,

Bagalá³

et al. 2015

J. Cancer

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“…Chloride intracellular channel proteins (CLIC), S100 proteins, tropomyosins (TPM), cathepsins, α-integrins (ITGA), eukaryotic elongation factors (EEF), and vimentin (VIM) were also discriminated through analyses on the RAS-modulated proteome of different types of cells investigated in previous studies [21]–[31]. The results obtained suggested that these could be important molecules participating in RAS-mediated carcinogenesis.…”

Section: Discussionmentioning

confidence: 93%

Proteome Analysis for Downstream Targets of Oncogenic KRAS - the Potential Participation of CLIC4 in Carcinogenesis in the Lung

et al. 2014

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This study investigated the proteome modulated by oncogenic KRAS in immortalized airway epithelial cells. Chloride intracellular channel protein 4 (CLIC4), S100 proteins (S100A2 and S100A11), tropomyosin 2, cathepsin L1, integrinsα3, eukaryotic elongation factor 1, vimentin, and others were discriminated. We here focused on CLIC4 to investigate its potential involvement in carcinogenesis in the lung because previous studies suggested that some chloride channels and chloride channel regulators could function as tumor suppressors. CILC4 protein levels were reduced in some lung cancer cell lines. The restoration of CLIC4 in lung cancer cell lines in which CLIC4 expression was reduced attenuated their growth activity. The immunohistochemical expression of the CLIC4 protein was weaker in primary lung cancer cells than in non-tumorous airway epithelial cells and was occasionally undetectable in some tumors. CLIC4 protein levels were significantly lower in a subtype of mucinous ADC than in others, and were also significantly lower in KRAS-mutated ADC than in EGFR-mutated ADC. These results suggest that the alteration in CLIC4 could be involved in restrictedly the development of a specific fraction of lung adenocarcinomas. The potential benefit of the proteome modulated by oncogenic KRAS to lung cancer research has been demonstrated.

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“…Dihydropyrimidinase‐related protein 3 is required for signalling by class 3 semaphorins and subsequent cytoskeleton remodelling. It is associated with neurofibromin, a negative regulator of Ras, indicating a role for DRP‐3 in proliferation and neural morphogenesis . Its regulation by retinoic acid also suggests a role in differentiation .…”

Section: Discussionmentioning

confidence: 99%

Proteomic analysis through larval development of Solea senegalensis flatfish

et al. 2015

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The post-embryonic development of the Senegalese sole, Solea senegalensis, a flatfish of growing interest in fisheries and aquaculture, is associated with drastic morpho-physiological changes during metamorphosis. Although in the last two decades knowledge on sole culture has notably increased, especially in Southern Europe, its progress was restricted due to lack of methods to control reproduction, improve larval quality and increase juvenile disease resistance. A limited knowledge of the physiological, molecular and genetic mechanisms involved is at the base of such limitation. A proteomic study was carried out to explore the molecular events that occur during S. senegalensis ontogenesis. Protein expression changes were monitored in larvae from 5 to 21 dph by combining 2DE and protein identification with de novo MS/MS sequencing. An average of 6177 ± 282 spots was resolved in 2DE gels. Hierarchical cluster analysis of the 705 selected spots grouped them in eight patterns. Thirty-four proteins were identified and assigned biological functions including structure, metabolism highlighting energy metabolism, transport, protein folding, stress response, chromatin organization and regulation of gene expression. These changes provide a sequential description of the molecular events associated with the biochemical and biological transformations that occur during sole larval development.

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Functional specific roles of H‐ras and N‐ras. A proteomic approach using knockout cell lines

Cited by 4 publications

References 63 publications

ERCC1 Induction after Oxaliplatin Exposure May Depend on KRAS Mutational Status in Colorectal Cancer Cell Line: In Vitro Veritas

ERCC1 Induction after Oxaliplatin Exposure May Depend on KRAS Mutational Status in Colorectal Cancer Cell Line: In Vitro Veritas

Proteome Analysis for Downstream Targets of Oncogenic KRAS - the Potential Participation of CLIC4 in Carcinogenesis in the Lung

Proteomic analysis through larval development of Solea senegalensis flatfish

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