Effects of Silencing RET/PTC1 Junction Oncogene in Human Papillary Thyroid Carcinoma Cells

Gilbert-Sirieix, Marie; Ripoche, Hugues; Malvy, Claude; Massaad-Massade, Liliane

doi:10.1089/thy.2010.0006

Cited by 14 publications

(11 citation statements)

References 26 publications

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“…Total RNAs of untreated and transfected cells were extracted using RNeasy mini-kit (Qiagen, Courtaboeuf, France). Protocol is detailed in Array Express (Accession Number: E-MTAB-2838) and previously described by Gilbert-Sirieix [ 22 ]. The mRNA was then labelled using fluorescent low-input linear amplification kit (Agilent Technologies, Massy, France).…”

Section: Methodsmentioning

confidence: 99%

Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

et al. 2015

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TMPRSS2-ERG junction oncogene is present in more than 50% of patients with prostate cancer and its expression is frequently associated with poor prognosis. Our aim is to achieve gene knockdown by siRNA TMPRSS2-ERG and then to assess the biological consequences of this inhibition. First, we designed siRNAs against the two TMPRSS2-ERG fusion variants (III and IV), most frequently identified in patients’ biopsies. Two of the five siRNAs tested were found to efficiently inhibit mRNA of both TMPRSS2-ERG variants and to decrease ERG protein expression. Microarray analysis further confirmed ERG inhibition by both siRNAs TMPRSS2-ERG and revealed one common down-regulated gene, ADRA2A, involved in cell proliferation and migration. The siRNA against TMPRSS2-ERG fusion variant IV showed the highest anti-proliferative effects: Significantly decreased cell viability, increased cleaved caspase-3 and inhibited a cluster of anti-apoptotic proteins. To propose a concrete therapeutic approach, siRNA TMPRSS2-ERG IV was conjugated to squalene, which can self-organize as nanoparticles in water. The nanoparticles of siRNA TMPRSS2-ERG-squalene injected intravenously in SCID mice reduced growth of VCaP xenografted tumours, inhibited oncoprotein expression and partially restored differentiation (decrease in Ki67). In conclusion, this study offers a new prospect of treatment for prostate cancer based on siRNA-squalene nanoparticles targeting TMPRSS2-ERG junction oncogene.

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Section: Methodsmentioning

confidence: 99%

Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

et al. 2015

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“…From a biological point of view, both siRNA RET/PTC (1 and 3) showed significant inhibitory effects on cell viability and on invasion/migration along with blockage of the cell cycle at G 0 /G 1 phase. Additionally, we observed apoptosis induction with caspase-3 and PARP1 cleavage [14,15]. Fig.…”

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confidence: 55%

Relevance of targeting RET/PTC junction oncogene and Wnt/β-catenin pathway in the treatment of papillary thyroid carcinoma: skill of 8-year work

Massaad-Massade

2014

Biopolym. Cell

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Papillary thyroid carcinoma (PTC) is the most common endocrine gland malignancy and occurs frequently due to the radiation exposure. PTC is characterized by the paracentric inversion in chromosome 10 leading to the fusion of RET with several genes present in thyroid named PTC. The RET/PTCs junction oncogenes are present in around 80 % of papillary thyroid carcinoma, the most frequent ones are RET/PTC1 and RET/PTC3. Interestingly, RET/PTCs are found only in the tumour cells and not in the surrounding normal tissues, therefore, they represent a good target for RNA interference strategies. We aimed, on the one hand, to inhibit dedifferentiation due to the RET/PTC junction oncogene by siRNA and, on the other hand, to investigate a role of Wnt/b-catenin pathway in the regulation of a tissue-specific transcription factor, the thyroid transcription factor-1 (TTF-1) essential for the differentiation of the thyroid. In this paper we summarised our main results obtained during eight years that pointed a new therapeutic strategy for papillary thyroid carcinoma.

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“…Four siRNAs were designed against RET/PTC3 , according to Reynolds et al, 2004 [26] and named as siRNA RET/PTC3 #1, #2, #3 and #4 (Table S1). The siRNA scrambled sequence was used as control (siRNA CT).…”

Section: Methodsmentioning

confidence: 99%

Effects of siRNA on RET/PTC3 Junction Oncogene in Papillary Thyroid Carcinoma: From Molecular and Cellular Studies to Preclinical Investigations

et al. 2014

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RET/PTC3 junction oncogene is typical of radiation-induced childhood papillary thyroid carcinoma (PTC) with a short latency period. Since, RET/PTC3 is only present in the tumour cells, thus represents an interesting target for specific therapy by small interfering RNA (siRNA). Our aim is to demonstrate in vitro and in vivo molecular and cellular effects of siRNA on RET/PTC3 knockdown for therapeutic application.First, we established a novel cell line stably expressing RET/PTC3 junction oncogene, named RP3 which was found tumorigenic in nude mice compared to NIH/3T3 mouse fibroblasts. Among four siRNAs and five concentrations tested against RET/PTC3, an efficient siRNA RET/PTC3 and an appropriate dose (50 nM) were selected which showed significant inhibition (p<0.001) of gene (RT-qPCR) and protein (Western blot) expressions. This siRNA was found efficient in RP3 cells (harbouring RET/PTC3) but non-efficient in BHP10-3 SCmice cell line (harbouring RET/PTC1) showing that a specific siRNA against fusion sequence is required to target the junction oncogene. In vitro siRNA RET/PTC3 showed significant (p<0.001) inhibitory effects on RP3 cell viability (MTT assay) and on invasion/migration (IncuCyte scratch test) with blockage of cell cycle at G0/G1 phase (flow cytometry) and induced apoptosis by caspase-3 and PARP1 cleavage (WB). After intravenous injection in nude mice, respective squalene (SQ) nanoparticles (NPs) of siRNA RET/PTC3 significantly (p<0.001) reduced RP3 tumour growth, oncogene and oncoprotein expressions, induced apoptosis and partially restored differentiation (decrease in Ki67). Hence, our findings highly support the use of siRNA RET/PTC3-SQ NPs as a new promising treatment for patients affected by PTC expressing RET/PTC3.

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Effects of Silencing RET/PTC1 Junction Oncogene in Human Papillary Thyroid Carcinoma Cells

Cited by 14 publications

References 26 publications

Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

Relevance of targeting RET/PTC junction oncogene and Wnt/β-catenin pathway in the treatment of papillary thyroid carcinoma: skill of 8-year work

Effects of siRNA on RET/PTC3 Junction Oncogene in Papillary Thyroid Carcinoma: From Molecular and Cellular Studies to Preclinical Investigations

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Effects of Silencing RET/PTC1 Junction Oncogene in Human Papillary Thyroid Carcinoma Cells

Cited by 14 publications

References 26 publications

Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

Relevance of targeting RET/PTC junction oncogene and Wnt/&beta;-catenin pathway in the treatment of papillary thyroid carcinoma: skill of 8-year work

Effects of siRNA on RET/PTC3 Junction Oncogene in Papillary Thyroid Carcinoma: From Molecular and Cellular Studies to Preclinical Investigations

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Relevance of targeting RET/PTC junction oncogene and Wnt/β-catenin pathway in the treatment of papillary thyroid carcinoma: skill of 8-year work