RESUMO
Introduction: NanoString nCounter technology, a novel molecular assay, is gaining prevalent use in clinical settings as it can overcome some common constraints that are associated with the use of polymerase chain reaction (PCR). Compared to PCR, NanoString technology does not involve any amplification steps, which significantly minimizes the chance of contamination. NanoString measures the number of mRNA transcripts directly by 'molecular counting', as up to 800 colored probes can be run simultaneously in a single reaction. Areas covered: This manuscript reviews the principle of NanoString and covers the main applications of NanoString in companion diagnostics with a focus on cancer immunotherapy and disease prognosis estimation. This review has also taken a step in the direction of personalized medicine, with the application of NanoString on the realm of companion diagnostics. Expert opinion: NanoString is going to take a vital role in companion diagnostics and personalized medicine, owing to its simple and easy to use characteristics. Yet, the use of NanoString requires normalization of expression level, which is represented by the copy number of respective mRNA, with a reference gene. Furthermore, difficulty in probe design, which demands prior knowledge of known sequence, has also been a limitation of NanoString.
Assuntos
Técnicas de Diagnóstico Molecular , Nanotecnologia , Nanomedicina Teranóstica/métodos , Biomarcadores , Perfilação da Expressão Gênica/métodos , Humanos , Medicina de Precisão/métodos , Medicina de Precisão/normas , Prognóstico , RNA Mensageiro/genética , Nanomedicina Teranóstica/normasRESUMO
Introduction: It is now clear that circulating cell-free ribonucleic acids (ccfRNAs), including messenger RNA (mRNA) and miRNA, are potential cancer biomarkers. As ccfmiRNA is relatively more stable than ccfmRNA, research should concentrate on developing novel methods to preserve the stability of ccfmRNA and standardization of the protocol which includes extraction, detection, and multicenter validation. Areas covered: This literature review concentrates on the potential of ccfRNA being used as a biomarker in cancer, with special focus on mRNAs and microRNAs (miRNAs). Expert opinion: With the advancement of high-throughput technologies such as RNA sequencing, a panel of biomarkers will be used for the diagnosis, prognosis and therapeutic monitoring of cancer patients. In order to achieve this important target, bioinformatics education to pathologists, scientists, and technologists in molecular diagnostic laboratories is essential. Moreover, the panel of these new ccfRNAs biomarkers has to obtain approval or clearance from an authority such as the US Food and Drug Administration (FDA), and the standard of utilizing these new protocols has to be recognized via accreditation exercise. Therefore, there is still a long way to go before an extensively use of ccfRNA biomarkers in cancer patients can be realized.
Assuntos
Biomarcadores Tumorais , Ácidos Nucleicos Livres , Neoplasias/diagnóstico , Neoplasias/genética , RNA , Detecção Precoce de Câncer/métodos , Detecção Precoce de Câncer/normas , Humanos , Biópsia Líquida/métodos , Biópsia Líquida/normas , MicroRNAs/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Neoplasias/sangue , Prognóstico , RNA Mensageiro/genéticaRESUMO
N16 is a protein from the nacreous layer of Pinctada fucata, a pearl oyster. It has been found to promote biomineralization, and we hypothesized that it also plays a role in bone metabolism. The cDNA of N16 was cloned and expressed in Escherichia coli to produce N16 protein, which was purified to high homogeneity by ion-exchange and gel filtration columns. The effects of N16 on osteoclast differentiation and osteogenesis were clarified using the murine preosteoclast cell line RAW 264.7 and the preosteoblast cell line MC3T3-E1. Results on preosteoclasts showed that N16 only slightly inhibited cell survival but significantly inhibited differentiation induced by receptor activator of nuclear factor kappa-B ligand (RANKL). Apart from reduced formation of multinucleated osteoclasts, N16-treated cells exhibited lower gene expression and enzymatic activity typical of mature osteoclasts. Actin ring formation and intracellular acidification essential for osteoclastic function were also impaired upon N16 treatment. At concentrations nontoxic to preosteoblasts, N16 strongly up-regulated alkaline phosphatase activity and increased mineralized nodule formation, which are indicative of differentiation into osteoblasts. These effects coincided with an increase in mRNA expression of osteoblast markers osteopotin and osteocalcin. The present study demonstrated that N16 has both anabolic and antiresorptive effects on bone, which makes it potentially useful for treating osteoporosis.
