TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours

Francesca Mastropasqua, Flaviana Marzano, Alessio Valletti, Italia Aiello, Giuseppe Di Tullio, Annalisa Morgano, Sabino Liuni, Elena Ranieri, Luisa Guerrini, Giuseppe Gasparre, Elisabetta Sbisà, Graziano Pesole, Antonio Moschetta, Mariano Francesco Caratozzolo, Apollonia Tullo

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: TRIM8 plays a key role in controlling the p53 molecular switch that sustains the transcriptional activation of cell cycle arrest genes and response to chemotherapeutic drugs. The mechanisms that regulate TRIM8, especially in cancers like clear cell Renal Cell Carcinoma (ccRCC) and colorectal cancer (CRC) where it is low expressed, are still unknown. However, recent studies suggest the potential involvement of some microRNAs belonging to miR-17-92 and its paralogous clusters, which could include TRIM8 in a more complex pathway. Methods: We used RCC and CRC cell models for in-vitro experiments, and ccRCC patients and xenograft transplanted mice for in vivo assessments. To measure microRNAs levels we performed RT-qPCR, while steady-states of TRIM8, p53, p21 and N-MYC were quantified at protein level by Western Blotting as well as at transcript level by RT-qPCR. Luciferase reporter assays were performed to assess the interaction between TRIM8 and specific miRNAs, and the potential effects of this interaction on TRIM8 expression. Moreover, we treated our cell models with conventional chemotherapeutic drugs or tyrosine kinase inhibitors, and measured their response in terms of cell proliferation by MTT and colony suppression assays. Results: We showed that TRIM8 is a target of miR-17-5p and miR-106b-5p, whose expression is promoted by N-MYC, and that alterations of their levels affect cell proliferation, acting on the TRIM8 transcripts stability, as confirmed in ccRCC patients and cell lines. In addition, reducing the levels of miR-17-5p/miR-106b-5p, we increased the chemo-sensitivity of RCC/CRC-derived cells to anti-tumour drugs used in the clinic. Intriguingly, this occurs, on one hand, by recovering the p53 tumour suppressor activity in a TRIM8-dependent fashion and, on the other hand, by promoting the transcription of miR-34a that turns off the oncogenic action of N-MYC. This ultimately leads to cell proliferation reduction or block, observed also in colon cancer xenografts overexpressing TRIM8. Conclusions: In this paper we provided evidence that TRIM8 and its regulators miR-17-5p and miR-106b-5 participate to a feedback loop controlling cell proliferation through the reciprocal modulation of p53, miR-34a and N-MYC. Our experiments pointed out that this axis is pivotal in defining drug responsiveness of cancers such ccRCC and CRC.

Original languageEnglish (US)
Article number67
JournalMolecular Cancer
Volume16
Issue number1
DOIs
StatePublished - Mar 21 2017

Fingerprint

Renal Cell Carcinoma
Colorectal Neoplasms
MicroRNAs
Cell Proliferation
Neoplasms
Heterografts
Pharmaceutical Preparations
cdc Genes
Cell Cycle Checkpoints
Luciferases
Protein-Tyrosine Kinases
Colonic Neoplasms
Transcriptional Activation
Western Blotting
Cell Line
Proteins

Keywords

  • Drug resistance
  • MiR-17 family
  • N-MYC
  • P53
  • TRIM8

ASJC Scopus subject areas

  • Molecular Medicine
  • Oncology
  • Cancer Research

Cite this

Mastropasqua, F., Marzano, F., Valletti, A., Aiello, I., Di Tullio, G., Morgano, A., ... Tullo, A. (2017). TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours. Molecular Cancer, 16(1), [67]. https://doi.org/10.1186/s12943-017-0634-7

TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours. / Mastropasqua, Francesca; Marzano, Flaviana; Valletti, Alessio; Aiello, Italia; Di Tullio, Giuseppe; Morgano, Annalisa; Liuni, Sabino; Ranieri, Elena; Guerrini, Luisa; Gasparre, Giuseppe; Sbisà, Elisabetta; Pesole, Graziano; Moschetta, Antonio; Caratozzolo, Mariano Francesco; Tullo, Apollonia.

In: Molecular Cancer, Vol. 16, No. 1, 67, 21.03.2017.

Research output: Contribution to journalArticle

Mastropasqua, F, Marzano, F, Valletti, A, Aiello, I, Di Tullio, G, Morgano, A, Liuni, S, Ranieri, E, Guerrini, L, Gasparre, G, Sbisà, E, Pesole, G, Moschetta, A, Caratozzolo, MF & Tullo, A 2017, 'TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours', Molecular Cancer, vol. 16, no. 1, 67. https://doi.org/10.1186/s12943-017-0634-7
Mastropasqua F, Marzano F, Valletti A, Aiello I, Di Tullio G, Morgano A et al. TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours. Molecular Cancer. 2017 Mar 21;16(1). 67. https://doi.org/10.1186/s12943-017-0634-7
Mastropasqua, Francesca ; Marzano, Flaviana ; Valletti, Alessio ; Aiello, Italia ; Di Tullio, Giuseppe ; Morgano, Annalisa ; Liuni, Sabino ; Ranieri, Elena ; Guerrini, Luisa ; Gasparre, Giuseppe ; Sbisà, Elisabetta ; Pesole, Graziano ; Moschetta, Antonio ; Caratozzolo, Mariano Francesco ; Tullo, Apollonia. / TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours. In: Molecular Cancer. 2017 ; Vol. 16, No. 1.
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T1 - TRIM8 restores p53 tumour suppressor function by blunting N-MYC activity in chemo-resistant tumours

AU - Mastropasqua, Francesca

AU - Marzano, Flaviana

AU - Valletti, Alessio

AU - Aiello, Italia

AU - Di Tullio, Giuseppe

AU - Morgano, Annalisa

AU - Liuni, Sabino

AU - Ranieri, Elena

AU - Guerrini, Luisa

AU - Gasparre, Giuseppe

AU - Sbisà, Elisabetta

AU - Pesole, Graziano

AU - Moschetta, Antonio

AU - Caratozzolo, Mariano Francesco

AU - Tullo, Apollonia

PY - 2017/3/21

Y1 - 2017/3/21

N2 - Background: TRIM8 plays a key role in controlling the p53 molecular switch that sustains the transcriptional activation of cell cycle arrest genes and response to chemotherapeutic drugs. The mechanisms that regulate TRIM8, especially in cancers like clear cell Renal Cell Carcinoma (ccRCC) and colorectal cancer (CRC) where it is low expressed, are still unknown. However, recent studies suggest the potential involvement of some microRNAs belonging to miR-17-92 and its paralogous clusters, which could include TRIM8 in a more complex pathway. Methods: We used RCC and CRC cell models for in-vitro experiments, and ccRCC patients and xenograft transplanted mice for in vivo assessments. To measure microRNAs levels we performed RT-qPCR, while steady-states of TRIM8, p53, p21 and N-MYC were quantified at protein level by Western Blotting as well as at transcript level by RT-qPCR. Luciferase reporter assays were performed to assess the interaction between TRIM8 and specific miRNAs, and the potential effects of this interaction on TRIM8 expression. Moreover, we treated our cell models with conventional chemotherapeutic drugs or tyrosine kinase inhibitors, and measured their response in terms of cell proliferation by MTT and colony suppression assays. Results: We showed that TRIM8 is a target of miR-17-5p and miR-106b-5p, whose expression is promoted by N-MYC, and that alterations of their levels affect cell proliferation, acting on the TRIM8 transcripts stability, as confirmed in ccRCC patients and cell lines. In addition, reducing the levels of miR-17-5p/miR-106b-5p, we increased the chemo-sensitivity of RCC/CRC-derived cells to anti-tumour drugs used in the clinic. Intriguingly, this occurs, on one hand, by recovering the p53 tumour suppressor activity in a TRIM8-dependent fashion and, on the other hand, by promoting the transcription of miR-34a that turns off the oncogenic action of N-MYC. This ultimately leads to cell proliferation reduction or block, observed also in colon cancer xenografts overexpressing TRIM8. Conclusions: In this paper we provided evidence that TRIM8 and its regulators miR-17-5p and miR-106b-5 participate to a feedback loop controlling cell proliferation through the reciprocal modulation of p53, miR-34a and N-MYC. Our experiments pointed out that this axis is pivotal in defining drug responsiveness of cancers such ccRCC and CRC.

AB - Background: TRIM8 plays a key role in controlling the p53 molecular switch that sustains the transcriptional activation of cell cycle arrest genes and response to chemotherapeutic drugs. The mechanisms that regulate TRIM8, especially in cancers like clear cell Renal Cell Carcinoma (ccRCC) and colorectal cancer (CRC) where it is low expressed, are still unknown. However, recent studies suggest the potential involvement of some microRNAs belonging to miR-17-92 and its paralogous clusters, which could include TRIM8 in a more complex pathway. Methods: We used RCC and CRC cell models for in-vitro experiments, and ccRCC patients and xenograft transplanted mice for in vivo assessments. To measure microRNAs levels we performed RT-qPCR, while steady-states of TRIM8, p53, p21 and N-MYC were quantified at protein level by Western Blotting as well as at transcript level by RT-qPCR. Luciferase reporter assays were performed to assess the interaction between TRIM8 and specific miRNAs, and the potential effects of this interaction on TRIM8 expression. Moreover, we treated our cell models with conventional chemotherapeutic drugs or tyrosine kinase inhibitors, and measured their response in terms of cell proliferation by MTT and colony suppression assays. Results: We showed that TRIM8 is a target of miR-17-5p and miR-106b-5p, whose expression is promoted by N-MYC, and that alterations of their levels affect cell proliferation, acting on the TRIM8 transcripts stability, as confirmed in ccRCC patients and cell lines. In addition, reducing the levels of miR-17-5p/miR-106b-5p, we increased the chemo-sensitivity of RCC/CRC-derived cells to anti-tumour drugs used in the clinic. Intriguingly, this occurs, on one hand, by recovering the p53 tumour suppressor activity in a TRIM8-dependent fashion and, on the other hand, by promoting the transcription of miR-34a that turns off the oncogenic action of N-MYC. This ultimately leads to cell proliferation reduction or block, observed also in colon cancer xenografts overexpressing TRIM8. Conclusions: In this paper we provided evidence that TRIM8 and its regulators miR-17-5p and miR-106b-5 participate to a feedback loop controlling cell proliferation through the reciprocal modulation of p53, miR-34a and N-MYC. Our experiments pointed out that this axis is pivotal in defining drug responsiveness of cancers such ccRCC and CRC.

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