Evaluation of biomarkers, genetic mutations, and epigenetic modifications in early diagnosis of pancreatic cancer

Figure 1 Distribution of samples (cases) based on subtypes of pancreatic cancer. PC: Pancreatic cancer.

Figure 2 Analysis of tumor biological marker (carbohydrate antigen 19-9, tissue polypeptide specific antigen, carcinoembryonic antigen, vascular endothelial growth factor-A, and epidermal growth factor receptor) in the serum of pancreatic cancer patients. PC: Pancreatic cancer; CA19-9: Carbohydrate antigen 19-9; TPS: Tissue polypeptide specific antigen; CEA: Carcinoembryonic antigen; VEGF-A: Vascular endothelial growth factor-A; EGFR: Epidermal growth factor receptor.

Figure 3 Mutational analysis of KRAS (Codon 12 and 13), DPC-4, and BRCA-2 gene mutations within pancreatic cancer cases. KRAS: Kirsten rat sarcoma; DPC-4: Deleted in pancreatic cancer-4; BRCA-2: Breast cancer type 2.

Figure 4 Representative agarose gel picture of polymerase chain reaction amplification (A) and restriction fragment length polymorphism using BstN1 (B) for KRAS codon 12. The arrow represents the 157 bp amplicon and M denotes the DNA marker (50 bp). Lane M represents a DNA marker (50 bp). Lanes 2, 4, 7, 8, and 10 represent the mutant band (undigested) of 157 bp. Lane 1, 3, 6, and 9 represent the wild band (digested) of 128 bp. U represents the undigested band used as mutant control.

Figure 5 Representative gel picture of polymerase chain reaction amplification (A) and restriction fragment length polymorphism using BglI (B) for K-RAS codon 13. The arrow represents the 157 bp amplicon and M denotes the DNA marker (100 bp). Lane M represents a DNA marker (100 bp). Lanes 2, 3, 6, 7, 8, 9, 10, and 11 represent the mutant band (undigested) of 157 bp. Lanes 1, 4, 5, and 8 represent a wild band (digested) of 125 bp.

Figure 6 Representative gel picture of polymerase chain reaction amplification and restriction fragment length polymorphism using MnlI of DPC-4. The arrow represents the 184bp amplicon and M denotes the DNA marker (100 bp). Lane M represents a DNA marker (50 bp). 117 bp and 67 bp represent the wild bands (digested). U represents the mutant control band of 184 bp (undigested).

Figure 7 Representative agarose gel picture of AS-polymerase chain reaction amplification (A) and gel picture representing polyacrylamide gel electrophoresis (20% gel) (B) of BRCA-2. All the lanes show 151 bp amplicon which is the wild band and M denotes the DNA marker (100 bp). M is the marker lane (25 bp). Here also, the only wild band (151 bp) is observed in all the lanes.

Figure 8 Gel picture representing MS-polymerase chain reaction for hMLH1 (A, 100 bp) and RASSF1A (B, 25 bp). M represents DNA marker; UMC represents unmethylated control; MC represents methylated control; NM represents normal methylated; NUM represents normal unmethylated; TUM represents tumor unmethylated; TM represents tumor methylation.

Figure 9 Methylation and unmethylation status in the promoter region of p16,RASSF1A, and hMLH1 within pancreatic cancer subjects. PC: Pancreatic cancer.

Figure 10  Interaction of KRAS, SMAD4 (DPC4), and BRCA2 with other genes based on various parameters. KRAS: Kirsten rat sarcoma; DPC-4: Deleted in pancreatic cancer-4; BRCA-2: Breast cancer type 2.

Figure 11  Interaction of MLH1, RASSF1, and CDKN2A (p16) with other important genes which have a role in the progression of pancreatic cancer.