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Copyright ©The Author(s) 2016.
World J Gastroenterol. Jul 7, 2016; 22(25): 5627-5641
Published online Jul 7, 2016. doi: 10.3748/wjg.v22.i25.5627
Table 1 Circulating tumor cells in pancreatic cancer
Ref.Patient characteristicsNumber of patients with PCaControlsEnrichment/isolation methodDetection methodDetection rate
Funaki et al[28], 1996Pre- or post-treatment9NANoneRT-PCR: CEA mRNA33.3% of PCa
Funaki et al[29], 1998Preoperative3NANoneRT-PCR: CEA-mRNA
Miyazono et al[30], 1999Intra-operative2115 HVDensity gradient enrichmentRT-PCR: CEA-mRNA61.9% of PCa
Chausovsky et al[33], 1999Metastatic PCa2822 BDDensity gradient enrichmentRT-PCR: CK20 mRNA78.6% of PCa
Z'graggen et al[124], 2001All stages, preoperative10566 HVDensity gradient enrichmentICC: CK-AE1/AE3Sensitivity: 26%, specificity: 96%
Lukyanchuk et al[34], 2003NA1118 HVDensity gradient enrichmentRT-PCR: CK-20 and PSCACK-20: 19 of 47 (40.4%), PSCA: 22 of 47 (46.8%)
Clarke et al[37], 2003NA1123 HVDensity gradient enrichmentRT-PCR: EGFR mRNA18% of PCa, 0.0% of HV
Mataki et al[32], 2004All stages2015 HV, 15 BDDensity gradient enrichmentRT-PCR: CEA mRNASensitivity: 75.0%, specificity: 94.6%
Zhang et al[35], 2005Stages II and III405 BD, 5 HVNot availableRT-PCR: CK20 mRNA57.5% of PCa
Soeth et al[36], 2005Preoperative154NADensity gradient enrichmentRT-PCR: CK20 mRNA33.8% of PCa
Ishizone et al[38], 2006All stages5510 CP, 70 HVDensity gradient enrichmentRT-PCR: a4GnT76.4% of PCa, 40.0% of CP, 17.1% of HV
Hoffmann et al[39], 2007All stages3716 CP, 15 BDDensity gradient enrichmentRT-PCR: CK-19 mRNA64% of PCa
Kurihara et al[41], 2008Stages II, III and IV2611 CP, 10 HVCELLSEARCH®42% of PCa
Zhou et al[40], 2011All stages2515 BDImmunomagnetic separationRT-PCR: C-MET, h-TERT, CK20, and CEASensitivity: 100%, specificity: 93.3%
Khoja et al[125], 2012Stages III and IV54 PCaNASize-based selectionRT-PCR: EpCAM, CK, vimentin, and CEAISET 49/54 (93%) vs CELLSEARCH® 21/54 (40%)
de Albuquerque et al[126], 2012Stages III and IV3440 HVImmunomagnetic separationRT-PCR: KRT19, MUC1, EpCAM, CEACAM5, and BIRC5Sensitivity: 47.1%, specificity: 100%
Kamande et al[127], 2013All stages125 HVMicrofluidic; ICCDAPI+, CD45-, CK+100% of PCa
Bidard et al[42], 2013Locally advanced PCa79NACELLSEARCH®11% of PCa
Iwanicki-Caron et al[43], 2013All stages40NASize-based selectionCell size and cytopathologic criteriaSensitivity: 55.5%, specificity: 100%, accuracy: 70%
Bobek et al[45], 2014All stages24NASize-based selectionDAPI, CK, CEA, vimentin IHC66.7% of PCa
Sheng et al[48], 2014Metastatic PCa18NAMultifluidic, "GEM"Chip94.4% of PCa
Rhim et al[47], 2014All stages1119 HVGeometrically enhanced differential immunocaptureICC for DAPI, CD45, CK, and PDX-173% of PCa
Catenacci et al[128], 2015Stages II, III and IV18NAImmune-magnetic separationCD45-negative and positive for CK8, -18, and/or -19 and DAPI118.4 ± 36.8 CTCs/7.5 mL PVB, compared with a mean of 0.8 ± 0.4 CTCs/7.5 mL PB
Earl et al[81], 2015All stages35NACELLSEARCH®20% of PCa
Cauley et al[44], 2015All stages1059 HVSize-based selectionCytomorphologic criteria48.6% of PCa
Kulemann et al[46], 2015Preoperative119 HVSize-based selection: ScreenCellCytologic and detection of KRAS mutation75% of early PCa, 71.4% of advanced PCa
Zhang et al[129], 2015All stages3230 HVICC and FISHDAPI+, CD45-, and CK+, or CEP8 > 2+Sensitivity: 63.6%, specificity: 94.4%
Bissolati et al[130], 2015Intra-operative20NACELLSEARCH®PVB: 40%, PB: 20%
Zhang et al[131], 20151515 HVImmunomagnetic separationBC-15 aptamer or anti-CK staining73.3% of PCa
Table 2 Circulating cell-free DNA in pancreatic cancer
Ref.PatientsControlsSampleMethodTarget candidateFindings
Shapiro et al[64], 1983201 MD185 BDSerumRadioimmunoassayCirculating DNA levelsSerum DNA concentration is markedly elevated in 90% of patients with PCa as compared with HV
Yamada et al[72], 199821 PCa-PlasmaMutant allele-specific amplification methodK-ras mutationIn 9 of 15 (60%) patients with K-ras gene mutation-positive tumors, an identical mutation was detected in the plasma DNA. Detection of K-ras mutations in plasma may be clinically useful for evaluating tumor burden and efficacy of treatment
Giacona et al[132], 19983 PCa3 HVPlasmaGel electrophoresis and measuring the variation in length by electron microscopyLengthThere are significant differences in non-cell-associated DNA in plasma between patients with PCa and HV
Theodor et al[73], 199920 PCa6 CP, 5 HVSerumPCRK-ras mutationK-ras gene mutations at codon 12 were detected in the sera of 14 of 20 patients with PCa and in none of the 6 patients with CP, or in the 5 HVs
Castells et al[74], 199944 PCa37 CPPlasmaRestriction fragment length polymorphism-PCR and single-strand conformation polymorphism techniquesK-ras mutationPlasma K-ras analysis is a highly specific, low-sensitivity approach that has diagnostic and prognostic clinical implications in patients with PCa
Zambon et al[75], 200029 PCa12 HVSerumME-PCRK-ras mutationK-ras was amplifiable in 2 patients with PCa (6.9%), and K-ras was not amplifiable in any of the 12 serum samples obtained from HVs
Maire et al[76], 200247 PCa31 CPSerumPCR and allele-specific amplificationKRAS2 mutationsKRAS2 mutations were found in 22 patients (47%) with PCa and in 4 controls with CP (13%) (P < 0.002)
Melnikov et al[65], 200930 PCa30 HVPlasmaMultiplexed array-mediated analysis of DNA methylationMethylationDifferential methylation profiling of plasma DNA can detect PCa with 76% sensitivity and 59% specificity
Liggett et al[66], 201030 PCa30 CP, 30 HVPlasmaMicroarray-mediated methylation analysisMethylationMethylation analysis achieved 81.7% sensitivity and 78% specificity (P < 0.01) in the detection of CP (HV vs CP) and 91.2% sensitivity and 90.8% specificity (P < 0.01) in the differential detection of PCa (PCa vs CP)
Chen et al[79], 201091 PCa-PlasmaDirect sequencingK-rasK-ras codon 12 mutations were found in 30 of 91(33%) plasma DNA samples and significantly reflected the clinical parameters, including TNM tumor staging and liver metastasis, and independent predict shorter survival time
Wu et al[80], 201424 PCa25 HVPlasmaCOLD-PCR combined with an unlabeled-probe HRM approachK-rasKRAS mutations were identified in 26 of 36 PCa cases (72.2%), but none were detected in the disease control and/or healthy group
Earl et al[81], 201531 PCa-PlasmaDigital PCRKRASKRAS mutant cfDNA was detected in 26% of patients at all stages, which correlated strongly with OS, 60 d for KRAS mutation-positive vs 772 days for KRAS mutation-negative patients
Zill et al[85], 201526 PCa-PlasmaSequenced on an Illumina Hi-Seq 2500KRAS, TP53, APC, FBXW7, and SMAD4The diagnostic accuracy of cfDNA sequencing was 97.7%, with 92.3% average sensitivity and 100% specificity across 5 informative genes
Singh et al[133], 2015PlasmaLevels of ctDNA and K-ras mutationHigher levels of plasma DNA were significantly associated with lower OS and advanced stage. However, k-ras mutation did not correlate with any of the clinicopathological parameters or survival
Kinugasa et al[82], 2015141 PCa20 CP, 20 HVSerumDigital PCRG12V, G12D, and G12R in codon 12 of K-ras geneK-ras mutation rate in ctDNA was 62.6%. The survival of patients with K-ras mutations in ctDNA was significantly shorter than that of patients without mutations
Sausen et al[83], 201577 PCa-PlasmaNext-generation sequencingThe 43% of patients with localized disease had detectable ctDNA at diagnosis. Detection of ctDNA after resection predicts clinical relapse and poor outcome, with recurrence by ctDNA detected 6.5 months earlier than with CT imaging
Takai et al[84], 2015259 PCa-PlasmaPicoliter-droplet digital PCR and targeted deep sequencingKRAS mutationKRAS mutations were identified in 14 of 48 patients (29.2%) examined by targeted deep sequencing of cfDNA
Table 3 Circulating noncoding RNA in pancreatic cancer
Ref.SampleCandidate targetPotential value
Wang et al[102], 2009PlasmamiR-210 (↑), miR-21 (↑), miR-155 (↑), miR-196a (↑)D
Ho et al[134], 2010PlasmamiR-210 (↑)D
Li et al[135], 2010PlasmamiR-200a (↑), miR-200b (↑)D
Ali et al[136], 2010PlasmamiR-21 (↑)D/P
Kong et al[105], 2011SerummiR-196a (↑)D/S/P
LaConti et al[137], 2011SerummiR-155 (↑)D
Morimura et al[138], 2011PlasmamiR-18a (↑)D
Liu et al[139], 2012SerummiR-16 (↑), miR-196a (↑)D
Liu et al[140], 2012SerummiR-20a (↑), miR-21 (↑), miR-24 (↑), miR-25 (↑),D/P
miR-99a (↑), miR-185 (↑), miR-191 (↑)
Li et al[141], 2012SerummiR-1290 (↑)D
Wang et al[142], 2013Whole bloodmiR-27a-3p (↑)D
Kawaguchi et al[143], 2013PlasmamiR-221 (↑), miR-375 (↓)D/S
Zhao et al[144], 2013SerummiR-192 (↑)D
Li et al[141], 2013SerummiR-1290 (↑)D
Wang et al[145], 2013SerummiR-21 (↑)T/P
Carlsen et al[146], 2013PlasmamiR-375 (↑)D
Que et al[147], 2013SerummiR-17-5p (↑), miR-21 (↑), miR-155 (↓), miR-196a (↓)D/S
Schultz et al[103], 2014Whole bloodMultigene indexD
Gao et al[148], 2014PlasmamiR-16 (↑)D
Chen et al[149], 2014PlasmamiR-182 (↑)D/S/P
Ganepola et al[150], 2015PlasmamiR-22 (↑), miR-642b (↑), miR-885-5p (↑)D
Abue et al[151], 2015PlasmamiR-21 (↑), miR-483-3p (↑)D/S/P
Slater et al[152], 2015SerummiR-196a (↑), miR-196b (↑)D
Kojima et al[104], 2015SerumMultigene indexD
Xu et al[153], 2015PlasmamiR-486-5p (↑), miR-938 (↑)D
Madhavan et al[154], 2015SerummiR-1246 (↑), miR-3976 (↑), miR-4306 (↑), miR-4644 (↑)D
Komatsu et al[123], 2015PlasmamiR-223 (↑)D/P
Miyamae et al[106], 2015PlasmamiR-744 (↑)D/S/P/T
Other noncoding RNAs
Wang et al[117], 2015PlasmaHOTTIP-005 (↑), RP11-567G11.1 (↑)D/P
Baraniskin et al[155], 2013Plasma/serumU2 snRNA (↑)D