Lack of Fas (APO-1/CD95) gene structural alterations or transcript variant ratio changes in breast cancer

Puiu, Liliana; Petrakou, Eftichia; Apostolidou, Anastasia; Athanassiadou, Aglaia; Psiouri, Lambrini; Papachatzopoulou, Adamantia; Gorgoulis, Vassilis G.; Kotsinas, Athanassios; Tzoracoeleftherakis, Evangelos; Maniatis, George M.; Voutsinas, Gerassimos E.

doi:10.1016/s0304-3835(03)00049-1

Cited by 8 publications

(8 citation statements)

References 29 publications

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“…Negative controls were performed with each RT-PCR reaction, omitting template. The efficiency of reverse transcription was verified by the detection of the GAPDH (glyceraldehyde phosphate dehydrogenase, Forward: 5′ -TGG TAT CGT GGA AGG ACT CAT GAC -3, Reverse: 5′ -ATG CCA GTG AGC TTC CCG TTC AG -3′), as previously described [15]. The products were resolved on a 2% agarose gel containing ethidium bromide and were visualized under ultraviolet light.…”

Section: A) Isolation Of Total Rna and Reverse Transcription-pcr (Rt-mentioning

confidence: 99%

RANKL and OPG mRNA Level after Non-Surgical Periodontal Treatment

et al. 2009

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Recent research evidence shows that the receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG) play an important role in osteoclastogenesis and the inflammatory bone loss during periodontitis. Bone remodeling process is dependent on the balance of these two proteins while a high ratio of RANKL/OPG characterizes the increased osteolytic process and it has been reported in inflammatory diseases including the periodontal disease. The purpose of this study was to determine the OPG and RANKL mRNA levels in periodontal tissues derived from patients with advanced chronic periodontitis after non-surgical periodontal therapy (SRP) and to compare the RANKL/OPG ration with that in healthy persons. Gingival biopsies were obtained from subjects with clinically healthy periodontium (H) (N = 11) and patients with advanced chronic periodontitis (CP) (N = 14). Total RNA was isolated from the gingival samples and 1 microg RNA was reverse transcribed to cDNA, followed by polymerase chain reaction (PCR) using specific primers for OPG and RANKL. The efficiency of reverse transcription was verified by the amplification of the GAPDH gene. The intensity of RT-PCR products was analyzed by a densitometer and was normalized to the intensity of the band for the housekeeping gene GAPDH. Immunohistochemical evaluation of the RANKL and OPG expression was also performed. The expression of RANKL as well as of OPG was reduced in CP specimens in comparison to that of healthy persons in a statistical significant way. However, the RANKL/OPG ratio showed to be slightly elevated in CP compared to H specimens but this finding was not of statistical significance. The immunohistochemical analysis revealed a non-uniform expression pattern for both proteins. Although further investigation is needed to identify the specific role of RANKL and OPG protein in periodontitis progression, our data after SRP might indicate the possible involvement of these proteins in the activation of pathways, which regulate the repair of the periodontal tissues.

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Section: A) Isolation Of Total Rna and Reverse Transcription-pcr (Rt-mentioning

confidence: 99%

RANKL and OPG mRNA Level after Non-Surgical Periodontal Treatment

et al. 2009

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“…Genomic DNA was extracted from the paraffin-embedded tissues, using a standard phenol-chloroform protocol and was subjected (200-700 ng DNA) to PCR amplification of FAS exon 9 using 2 different primer pairs, as previously described [12]. A SSCP analysis followed, each time using 2 different conditions and temperatures.…”

Section: Polymerase Chain Reaction -Single Strand Conformation Polymomentioning

confidence: 99%

“…All RT reactions were performed independently of the PCR reactions. Amplification of FAS cDNA by PCR [12] was followed by a second round of semi-nested PCR. The amplification reaction included 2 μL cDNA template, plus 10 pmol/μL primers, 10 pmol/μL dNTPs, 1 × Buffer, 1.5 mM MgCl2 and 1.5 U Platinum Taq polymerase (Invitrogen, Life Technologies) in a final volume of 50 μL.…”

Section: Isolation Of Total Rna and Reverse Transcription-pcrmentioning

confidence: 99%

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Resistance to Fas-Mediated Apoptosis Does Not Correlate to Structural Alterations or Expression Changes of the Death Receptor in Papillary Thyroid Carcinomas

et al. 2018

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Background: Malignant cells exhibit significant resistance to FAS-mediated cell death, through different processes, including FAS mutations, soluble FAS expression, or FAS transcriptional dysregulation by P53, eventually escaping from immune surveillance. Since thyroid carcinomas were shown to be resistant to FAS-mediated apoptosis, we investigated the above mechanisms in thyroid carcinoma samples. Methods: Thirty-seven thyroid carcinoma samples were analyzed for mutations in FAS exon 9 and TP53 exons 5–8 and protein expression by means of immunohistochemistry. Moreover, thyroid carcinoma mRNA samples were subjected to reverse transcription – PCR, to evaluate the relative expression of transmembrane FAS versus its soluble form. Results: Analysis revealed indications for TP53 mutations in the anaplastic carcinomas, but not in the other thyroid specimens examined for TP53 or FAS exon 9 mutations. FAS receptor expression was observed in almost all thyroid specimens (97%) with significant up-regulation in papillary carcinomas. P53 nuclear staining was observed only in anaplastic carcinomas. Full-length FAS mRNA was detected in all specimens examined, with soluble FAS mRNA being either absent or present in very low amounts. Conclusions: Our results denote that FAS death domain or TP53 DNA-binding domain mutations, down-regulation of FAS receptor expression, or expression of FAS soluble isoform are not responsible for the seeming inhibition of FAS-mediated apoptosis in papillary thyroid carcinoma cells.

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“…14 The products were resolved on a 2% agarose gel containing ethidium bromide and were visualized under ultraviolet light.…”

Section: Studies On Cell Lines and Culturesmentioning

confidence: 99%

Fibrillin expression and localization in various types of carcinomas of the thyroid gland

et al. 2006

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Fibrillin is an extracellular matrix (ECM) glycoprotein, a main component of microfibrills, suggested to support cell attachment and to impact cell differentiation and migration. The aim of this study was to investigate fibrillin-1 expression in thyroid carcinomas at mRNA and protein level, since ECM proteins are suggested to be of great importance for the metastatic potential of carcinomas. RNA was extracted from 13 thyroid carcinoma cell lines and RT-PCR analysis with gene-specific primers revealed fibrillin-1 mRNA expression in all cell lines, with highest expression in the follicular carcinoma cell line WRO and lowest expression in the two anaplastic cell lines (APO, FRO). Furthermore, we investigated fibrillin-1 expression by immumohistochemistry in a commercially available tissue microarray including 50 thyroid carcinomas as well as in archival tissue from 33 thyroid carcinomas. Fibrillin-1 demonstrated a cytoplasmic location in the neoplastic cells of almost all carcinomas apart from the follicular ones. The most intense staining was observed in papillary carcinomas with some evidence of a slight increased intensity in advanced stages. Our data indicate that fibrillin-1 is strongly expressed by the neoplastic cells of thyroid carcinomas in different degree in the various histologic types and might be implicated in cell-stroma interaction in terms of signaling, attachment and migration.

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Lack of Fas (APO-1/CD95) gene structural alterations or transcript variant ratio changes in breast cancer

Cited by 8 publications

References 29 publications

RANKL and OPG mRNA Level after Non-Surgical Periodontal Treatment

RANKL and OPG mRNA Level after Non-Surgical Periodontal Treatment

Resistance to Fas-Mediated Apoptosis Does Not Correlate to Structural Alterations or Expression Changes of the Death Receptor in Papillary Thyroid Carcinomas

Fibrillin expression and localization in various types of carcinomas of the thyroid gland

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