Liver cancer-specific biomarker
A biomarker, liver cancer technology, applied in the fields of biomaterial analysis, biochemical equipment and methods, and microbial determination/inspection.
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Embodiment 1
[0061] Example 1. Screening of blood markers using the database
[0062] 1-1. Screening of liver cancer cell-specific markers
[0063] Blood samples (15 normal liver patient samples (Normal liver, NL); 20 chronic hepatitis patient samples (Chronic hepatitis, CH); 10 3 samples from patients with liver cirrhosis (Liver Cirrhosis, LC); 18 samples from patients with early-stage hepatocellular carcinoma (Early HCC, eHCC); and 45 samples from patients with advanced hepatocellular carcinoma (Advanced HCC, avHCC)) After extracting total RNA using TRIzol reagent, the sequencing library was prepared using RNA Library Prep Kit for Illumina (Cat#E7420L), and sequenced by Illumina HiSeq 2000 according to the standard method of Illumina Company. After using STAR and Gencode v.25 to map the entire transcriptome of the analyzed liver, the expression profiles were replaced by FPKM values, then Gencode v.25 was used to classify the genotypes, and SignalP 4.1 was used to derive 12654 signal p...
Embodiment 2
[0068] Example 2. ELISA analysis of selected markers for the first time
[0069] 2-1. Confirmation of marker expression profiles
[0070] Blood samples (135 samples from 16 normal liver patients (Normal liver, NL ); 65 samples from 13 patients with chronic hepatitis (Chronic hepatitis, CH); 103 samples from 15 patients with liver cirrhosis (LiverCirrhosis, LC); 227 samples from 35 patients with early-stage hepatocellular carcinoma (Early HCC , eHCC); and 241 samples from 24 patients with advanced hepatocellular carcinoma (Advanced HCC, avHCC)) ( Figure 7 ), the expression levels of the selected 10 marker genes ( Figures 8a-8b ).
[0071] As a result, in the case of CCNB2, an average of 0.02 ng / ml was measured in patients with normal liver (NL), 0.2029 ng / ml in patients with chronic hepatitis (CH), and 0.43 ng / ml in patients with cirrhosis (LC), Patients with early stage hepatocellular carcinoma (eHCC) exhibited 0.27 ng / ml, and patients with advanced hepatocellular carc...
Embodiment 3
[0075] Example 3. Verification of early cancer diagnostic markers HMMR, NXPH4, PITX1, THBS4 and UBE2T
[0076] 3-1. Confirmation of marker expression profiles
[0077] Blood samples (222 samples from 49 normal liver patients (Normal liver, NL ); 115 samples from 31 patients with chronic hepatitis (Chronic hepatitis, CH); 183 samples from 46 patients with liver cirrhosis (LiverCirrhosis, LC); 345 samples from 77 patients with early-stage hepatocellular carcinoma (Early HCC , eHCC); and 283 samples from 64 patients with advanced hepatocellular carcinoma (Advanced HCC, avHCC)) ( Figure 10 ), the expression levels of the selected 10 marker genes ( Figures 11a-11b ).
[0078] As a result, when the protein expression levels of the five markers were measured in the validation cohort, in the case of HMMR, by comparing the results of the normal group with each liver disease group among a total of 230 samples, it was shown that in addition to Except for the liver cirrhosis group...
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