MicroRNA/gene profiling unveils early molecular changes and nuclear factor erythroid related factor 2 (NRF2) activation in a rat model recapitulating human hepatocellular carcinoma (HCC)

Petrelli, Annalisa; Perra, Andrea; Corà, Davide; Sulas, Pia; Menegon, Silvia; Manca, Claudia; Migliore, Cristina; Kowalik, Marta Anna; Ledda-Columbano, Giovanna M.; Giordano, Silvia; Columbano, Amedeo

doi:10.1002/hep.26616

Cited by 111 publications

(124 citation statements)

References 31 publications

Supporting

Mentioning

114

Contrasting

Unclassified

Order By: Relevance

“…Neoplastic hepatocytes were isolated by collagenase perfusion from the liver of a rat with multiple HCCs generated by the R-H model (R-H cells). 12 In agreement with our in vivo data, R-H cells bear a mutation of Nrf2 (D38G). To establish the biological effect of the modulation of Nrf2 levels in HCC cells, we performed soft agar assays on Nrf2-silenced cells (Supporting Fig.…”

Section: Resultssupporting

confidence: 89%

“…3C), suggesting that at late stages the Nrf2/Keap1 pathway no longer depends on the presence of activating mutations only but can be activated by other mechanisms as well. 12 The Nrf2/Keap1 Pathway Is Also Activated in Lung Metastases. Finally, we also examined the Nrf2/Keap1 pathway in lung metastases of rats sacrificed 14 months after DENA administration.…”

Section: Resultsmentioning

confidence: 99%

“…We isolated RNA from microdissected preneoplastic and neoplastic lesions using MirVana (Ambion, Austin, TX), as reported. 12 Retrotranscription was performed starting from 0.5 lg RNA/sample using the High Capacity Kit (Applied Biosystems, Carlsbad, CA). Gene expression was assessed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) using specific TaqMan probes (Nqo1, Gclc, Gsta4, Gs, Nrf2, Gapdh, and bactin; Applied Biosystems) or Express Sybr Green (Invitrogen, Paisley, UK) for Keap1 and b-actin.…”

Section: Methodsmentioning

confidence: 99%

“…12 Stable Transduction With Nrf2 Targeting Short Hairpin RNA. Cells isolated from an HCC-bearing rat (resistant-hepatocyte [R-H] cells), 12 cultured in Roswell Park Memorial Institute 10% fetal bovine serum medium, were stably transduced with the 217CS-SH213J-LVRH1P lentiviral vector containing a specific custom-designed Nrf2 targeting short hairpin RNA (GeneCopoeia). Transduced cells were selected by treatment with puromycin (1 lg/mL for 7 days; Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…12 Therefore, this study investigated whether, similar to the human situation, mutations of Nrf2 and/or Keap1 could be responsible for the alteration of this pathway. Using the same animal model, we also investigated the occurrence of Ctnnb1 mutations in the different stages of hepatocarcinogenesis.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Nrf2, but not β‐catenin, mutation represents an early event in rat hepatocarcinogenesis

et al. 2015

Self Cite

View full text Add to dashboard Cite

Hepatocellular carcinoma (HCC) develops through a multistage process, but the nature of the molecular changes associated with the different steps, the very early ones in particular, is largely unknown. Recently, dysregulation of the NRF2/KEAP1 pathway and mutations of these genes have been observed in experimental and human tumors, suggesting their possible role in cancer development. To assess whether Nrf2/Keap1 mutations are early or late events in HCC development, we investigated their frequency in the rat Resistant Hepatocyte model, consisting of the administration of diethylnitrosamine followed by a brief exposure to 2-acetylaminofluorene. This model enables the dissection of all stages of hepatocarcinogenesis. We found that Nrf2/Keap1 mutations were present in 71% of early preneoplastic lesions and in 78.6% and 59.3% of early and advanced HCCs, respectively. Mutations of Nrf2 were more frequent, missense, and located in the Nrf2-Keap1 binding region. Mutations of Keap1 occurred at a much lower frequency in both preneoplastic lesions and HCCs and were mutually exclusive with those of Nrf2. Functional in vitro and in vivo studies showed that Nrf2 silencing inhibited the ability of tumorigenic rat cells to grow in soft agar and to form tumors. Unlike Nrf2 mutations, those of Ctnnb1, which are frequent in human HCC, were a later event as they appeared only in fully advanced HCCs (18.5%). Conclusion: In the Resistant Hepatocyte model of hepatocarcinogenesis the onset of Nrf2 mutations is a very early event, likely essential for the clonal expansion of preneoplastic hepatocytes to HCC, while Ctnnb1 mutations occur only at very late stages. Moreover, functional experiments demonstrate that Nrf2 is an oncogene critical for HCC progression and development. (HEPATOLOGY 2015;62:851-862) See Editorial on Page 677 H epatocellular carcinoma (HCC) is the third most frequent cause of cancer-related deaths worldwide. 1 Unfortunately, our knowledge of the genomic alterations implicated in HCC initiation and progression is still fragmentary. Moreover, different from other solid tumors, no driving genetic alteration to which tumor cells are addicted and that can be effectively targeted has ever been identified. In this scenario, HCC is probably one of the tumor types where a more complete understanding of the underlying genetic alterations could have a major impact on the development of new treatment strategies. Also known as NFE2L2, NRF2 is of particular interest as conflicting results have been reported concerning the role of the NRF2/KEAP1 pathway in cancer development. 2 It is a master transcriptional activator of genes encoding enzymes that protect cells from oxidative stress and

show abstract

Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Nrf2, but not β‐catenin, mutation represents an early event in rat hepatocarcinogenesis

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

Local hypothyroidism favors the progression of preneoplastic lesions to hepatocellular carcinoma in rats

et al. 2014

Self Cite

View full text Add to dashboard Cite

Thyroid hormone receptors (TRs) are ligand-dependent transcription factors that mediate most of the effects elicited by the thyroid hormone, 3,5,3 0 -L-triiodothyronine (T3). TRs have been implicated in tumorigenesis, although it is unclear whether they act as oncogenes or tumor suppressors, and at which stage of tumorigenesis their dysregulation occurs. Using the resistant-hepatocyte rat model (R-H model), we found downregulation of TRb1 and TRa1 and their target genes in early preneoplastic lesions and hepatocellular carcinoma (HCCs), suggesting that a hypothyroid status favors the onset and progression of preneoplastic lesions to HCC. Notably, TRb1 and, to a lesser extent, TRa1 down-regulation was observed only in preneoplastic lesions positive for the progenitor cell marker, cytokeratin-19 (Krt-19) and characterized by a higher proliferative activity, compared to the Krt-19 negative ones. TRb1 down-regulation was observed also in the vast majority of the analyzed human HCCs, compared to the matched peritumorous liver or to normal liver. Hyperthyroidism induced by T3 treatment caused up-regulation of TRb1 and of its target genes in 1 preneoplastic rat lesions and was associated with nodule regression. In HCC, TRb1 down-regulation was not the result of hypermethylation of its promoter, but was associated with an increased expression of TRb1-targeting microRNAs ([miR]-27a, -181a, and -204). An inverse correlation between TRb1 and miR-181a was also found in human cirrhotic peritumoral tissue, compared to normal liver. Conclusion: Down-regulation of TRs, especially TRb1, is an early and relevant event in liver cancer development and is species and etiology independent. The results also suggest that a hypothyroid status of preneoplastic lesions may contribute to their progression to HCC and that the reversion of this condition may represent a possible therapeutic goal to interfere with the development of this tumor. (HEPATOLOGY 2015;61:249-259) T he thyroid hormones, 3,5,3 0 -triiodo-L-thyronine (T3) and thyroxine (T4), influence a variety of physiological processes.1,2 Most of the effects of T3 are mediated by thyroid hormone nuclear receptors (TRs), which are members of the steroid/thyroid receptor superfamily of nuclear hormone receptors, acting as transcription factors.

show abstract

Downregulation of indolethylamine N‐methyltransferase is an early event in the rat hepatocarcinogenesis and is associated with poor prognosis in hepatocellular carcinoma patients

López-Torres

Torres-Mena

Castro-Gil

et al. 2022

The Journal of Gene Medicine

View full text Add to dashboard Cite

Hepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide, often preceded by cirrhosis and usually diagnosed at advanced stages; therefore, identifying molecular changes at early stages is an attractive strategy for detection and timely treatment. Here, we investigated the progressive transcriptomic changes during experimental hepatocarcinogenesis to identify novel early tumor markers in an HCC model induced by chronic administration of sublethal doses of diethylnitrosamine. An

show abstract

MicroRNA/gene profiling unveils early molecular changes and nuclear factor erythroid related factor 2 (NRF2) activation in a rat model recapitulating human hepatocellular carcinoma (HCC)

Cited by 111 publications

References 31 publications

Nrf2, but not β‐catenin, mutation represents an early event in rat hepatocarcinogenesis

Nrf2, but not β‐catenin, mutation represents an early event in rat hepatocarcinogenesis

Local hypothyroidism favors the progression of preneoplastic lesions to hepatocellular carcinoma in rats

Downregulation of indolethylamine N‐methyltransferase is an early event in the rat hepatocarcinogenesis and is associated with poor prognosis in hepatocellular carcinoma patients

Contact Info

Product

Resources

About