Table 4.
Detection of pancreatic cancer mutations, methylations and miRNA changes in stool.
| Reference | Study Population | Controls | Methods | DNA-/RNA-Based Markers | Detection Rate in Stool/ST and SF |
Detection Rate in TT/PT/PJ |
Other Markers | Main Findings/Authors’ Conclusions |
|---|---|---|---|---|---|---|---|---|
| Caldas et al., 1994 [95] | 11 PDAC, 3 CCA, 3 CP, 1 PTu |
n.a. | Plaque hybr. assay | KRAS codon 12 in stool and tissue | In 55% of PDAC, in 67% of CCA, in 33% of CP. | TT/PT: in 100% of PDAC, 67% of CCA, 65% of duct lesions. | n.a. | KRAS mutations from PC cells and from abnormal duct epithelium can be detected in stool; potential use for screening of PDAC and precursor lesions. |
| Berndt et al., 1998 [96] | 42 PDAC, 1 CAC, 1 CA, 7 PAC, 1 NEC, 2 PI, 7 CP |
6 HC | Mut.-enr. PCR and rev. dot-plot hybr. in microplates | KRAS in stool and tissue | In 40% of PDAC, in 100% of CAC, in 33% of CP; ST 42.3%, SP 66.7%. |
TT/PT: in 91% of PDAC, 71% of PAC, 67% of CP. | Serum CA 19–9 and CEA |
Diagnostic ST of KRAS in stool is only 40%, which is similar to CEA but much lower than CA 19–9. Establishment of marker combinations for stool testing is necessary. |
| Wenger et al., 1999 [97] | 36 PDAC, 7 PAC, 1 CAC, 2 PI, 5 CP | 10 HC | Allele-specific capture probes, mut.-enr. PCR | KRAS in stool and tissue | In 20% of PDAC, in 100% of CAC, in 40% of CP. | TT/PT: in 78% of PDAC, 100% of CAC, 14% of PAC, 20% of CP. |
Serum CA 19–9 and CEA |
Mut. KRAS analysis in tissue did not distinguish between benign and malignant pancreatic disease. Only 20–40% of PC cases can be traced back from stool samples. Stool analysis could still be useful to detect more cases and increase survival. |
| Pezzilli et al., 2006 [98] | PDAC, CAC, PET, CP, pseudocysts, benign congenital pancreatic mass | n.a. | PCR amplification | KRAS codon 12 in stool and blood | No detection | No detection | n.a. | KRAS mutation analysis in blood and stool is not useful for differentiating benign and malignant pancreatic masses. Further studies are needed to find simple and useful genetic markers for the detection of pancreatic malignancy. |
| Lu et al., 2002 [99] | 201 PC or BPD | 60 HC | PCR-RFLP, PCR-SSCP | KRAS and TP53 in stool and PJ | Mut. KRAS in 88% of PC, 51.1% of BPD, 19.6% of HC; mut. TP53 in 37.1% of PC and 19.1% of CP. | PJ: mut. KRAS in 87.8% and 23.5%, mut. TP53 in 47.4% and 12.5% of PC and BPD. | n.a. | KRAS mutation analysis in pancreatic juice might be used in PC diagnosis. Combined KRAS and TP53 mutation analysis in stool can improve PC screening. |
| Wu et al., 2006 [100] | 31 PC for fecal analysis, 48 PC for serum analysis | 85 controls with benign digestive disorders | PCR-RFLP, PCR-SSCP | KRAS and TP53 in stool | Mut. KRAS in 77.4% of PC and 18.2% of controls; mutated TP53 in 25.8% of PC and 4.71% of controls. | n.a. |
Serum CA 19–9, CA 242, CA 50, CEA |
Fecal KRAS and TP53 mutations do not differ between tumor subgroups, which indicates an early role in tumorigenesis. The diagnostic value of CA 19–9 and CA 242 could be improved by combination with fecal KRAS analysis. |
| Hwang et al., 2011 [101] | 14 PC, 6 IPMN | 20 HC | Hybrid capture enrichment of KRAS; QuARTS | KRAS in stool | 62% ST for PC and 83% for IPMN (at 90% SP cutoff). | n.a. | n.a. | Pancreatic neoplasia can be detected by stool screening, but further studies using genetic and epigenetic alterations complementary to KRAS are needed. |
| Wang et al., 2018 [102] | 88 PC, 35 CP, 19 BPD |
3 HC | Magnetic nanoparticle trace capture probe and PCR | KRAS in stool and tumor tissue | Mut. KRAS in 81.8% of PC and 18.5% of BPD, 0% of HC; ST and SF for detecting PC: 81.8% and 81.5%. | n.a. | Serum CA 19–9 |
ST and SF of fecal mut. KRAS for detection of PC was slightly higher than that of serum CA 19–9. By combining both markers, sensitivity could be increased to 97.9% while specificity stayed the same. |
| Haug et al., 2007 [103] | 875 unselected older adults |
n.a. | Mut.-enr. PCR and allele-specific hybr. reaction | KRAS codons 12 and 13 in stool | 8% overall prevalence of mut. KRAS. | n.a. | n.a. | Tentative association between decreased fecal pancreatic elastase 1 and mut. KRAS in stool, but no KRAS mutations detected in cases that later developed CRC. This assay could be used for early detection of PC, but not for CRC screening. |
| Kisiel et al., 2012 [92] | 58 PDAC | 65 HC | Sequence specific gene capture (stool), MSP (tissue and stool); QuARTS | Mut. KRAS and meth. BMP3, NDRG4, EYA4, UCHL1, MDFI, Vimentin, CNTNAP2, SFRP2, TFPI2 in stool and tissue | Meth. BMP3 detected 51%, mut. KRAS detected 50% and combination of both detected 67% of PDAC. | n.a. | n.a. | PC can be detected from stool assay of methylated gene markers; BPM3 performed well alone; combining it with mut. KRAS increased detection rate for PDAC. |
| Link et al., 2012 [106] | 15 PC, 15 CP | 15 HC | Taq-Man miRNA assay | miR-21, -143, -155, -196a, -210, -216a, -375 |
Lower expression of miR-216a, -196a, -143 and -155 in PC compared to HC. | n.a. | n.a. | Differentially expressed miRNAs can be detected in stool of PC patients. This may be used as biomarker for PC screening. |
| Ren et al., 2012 [107] | 29 PC, 22 CP | 13 HC | Taq-Man miRNA assay | miR-16, -21, -155, -181a, -181b, -196a and -210 | mi-RNAs discriminated PC from HC; miR-181b at a ST and SF of 84.6% and 51.7%; miR-210 at a ST and SF of 84.6% and 65.5%. | n.a. | n.a. | Fecal miRNAs may be used as novel biomarkers for PC screening. |
| Yang et al., 2014 [108] | 30 PDAC, 10 CP | 15 HC | miRNA expr. analysis with qRT-PCR | miR-21, -155, -196a, -216 and -217 | Sign. higher expr. of miR-21 and -155 and lower expr. of miR-216 in PC compared to HC; ST of miR-21 and -155: 93.33%; ST and SF of miR-21, -155 and -216: 83.33% and 83.33%. | TT/PT/PJ: sign. higher expr. of miR-21 and -155; sign. lower expr. of miR-216 in PDAC compared to CP. | n.a. | MiRNA stool sampling and analysis is highly reproducible. Consistency in expression levels of miR-21, -155 and -216 in matched PC tissue, PJ, and stool samples. Combination of two or three miRNA markers yields enough ST and SF for their possible use as biomarkers for PC screening. |
BPD = benign pancreatic disease, CA = cystadenoma, CAC = cystadenocarcinoma, CP = chronic pancreatitis, CRC = colorectal cancer, expr. = expression, HC = healthy controls, hybr. = hybridization, MSP = real-time methylation-specific PCR, meth. = methylated, mut. = mutant, mut.-enr. = mutant-enriched, NEC = neuroendocrine carcinoma, n.a. not applicable, PA = papillary adenocarcinoma, PAC = periampullary carcinoma, PC = pancreatic cancer, PDAC = pancreatic ductal adenocarcinoma, PET = pancreatic endocrine tumor, PI = pancreatic insulinoma, PJ = pancreatic juice, PT = pancreatic tissue, PTu = pancreatic tuberculosis, QuARTS = quantitative allele-specific real-time target and signal amplification, qRT-PCR = quantitative Reverse Transcription PCR, RFLP = restriction fragment length polymorphisms, SF = specificity, sign. = significantly, SSCP = single-strand conformation polymorphism, ST = sensitivity, TT = tumor tissue.