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Fine-needle aspiration technique under endoscopic ultrasound guidance:A technical approach for RNA profiling of pancreatic neoplasms

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World Journal of Gastrointestinal Oncology 2024年第6期16卷 2663-2672页

作者： Sabina Sherafedinovna Seyfedinova Olga Aleksandrovna Freylikhman polina sergeevna sokolnikova Konstantin Aleksandrovich Samochernykh Anna Aleksandrovna Kostareva Olga Viktorovna Kalinina Evgeniy Gennadievich Solonitsyn Research Laboratory of Digestive Cancer Almazov Medical Research CentreSaint-Petersburg 197341Russia Research Laboratory of Molecular and Cellular Modeling and Gene Therapy Almazov Medical Research CentreSaint-Petersburg 197341Russia Head of Russian Neurosurgical Institute Named after Prof.A.L.Polenova Russian Neurosurgical Institute Named after Prof.A.L.PolenovaSaint-Petersburg 191014Russia Head of Institute of Molecular Biology and Genetics Almazov National Medical Research CentreSaint-Petersburg 197341Russia Research Laboratory of Autoimmune and Autoinflammatory Diseases Almazov National Medical Research CentreSaint-Petersburg 197341Russia

BACKGROUND Early diagnosis of pancreatic ductal adenocarcinoma(PDAC)has been a longstanding *** prognosis of patients with PDAC depends on the stage at *** is necessary to identify biomarkers for the detection and differentiation of pancreatic tumors and optimize PDAC sample preparation procedures for DNA and RNA *** molecular studies are done using paraffin-embedded blocks;however,the integrity of DNA and RNA is often compromised in this ***,RNA isolated from human pancreatic tissue samples is generally of low quality,in part,because of the high concentration of endogenous pancreatic RNAse activity *** To assess the potential of endoscopic ultrasound-guided fine-needle aspiration(EUS-FNA)to obtain specimens from pancreatic neoplasms for subsequent RNA molecular profiling,including next-generation sequencing(NGS).METHODS Thirty-four EUS-FNA samples were included in this study:PDAC(n=15),chronic pancreatitis(n=5),pancreatic cysts(n=14),mucinous cysts(mucinous cystic neoplasia/intraductal papillary mucinous neoplasia)n=7,serous cystic neoplasms n=5,and pseudocysts n=*** material consisted of cyst fluid and cyst wall samples obtained by through-the-needle biopsy(TTNB).Samples were stored at -80℃ until *** purity(A260/230,A260/280 ratios),concentration,and integrity(RIN)were ***-time polymerase chain reaction was conducted on all samples,and small RNA libraries were prepared from solid mass *** RNA was successfully extracted from 29/34(85%)EUS-FNA samples:100% pancreatic adenocarcinoma samples,100% chronic pancreatitis samples,70% pancreatic fluid cyst samples,and 50%TTNB *** relative expression of GAPDH and HPRT were obtained for all successfully extracted RNA samples(n=29)including lowquality RNA *** concentration and nonoptimal RIN values(no less than 3)of RNA extracted from EUS-FNA samples did not prevent NGS library *** suitability of cyst fluid samples

关键词： Endoscopic ultrasound-guided,fine-needle aspiration Pancreatic cancer Pancreatic cysts RNA extraction Through-the-needle biopsy Next-generation sequencing

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