Methylated biomarkers related to gastric cancer lymph node metastasis detection or combination and application thereof
A technology of biomarkers and lymph node metastasis, which is applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve the problem of limited accuracy of lymph node status assessment in gastric cancer, and achieve accurate clinical diagnosis and guidance The effect of treatment
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Embodiment 1
[0044] Example 1 Biomarker (marker) screening
[0045] 1. Sample source:
[0046] Samples must meet the following requirements: 1) patients with early gastric cancer diagnosed as T1 stage; 2) no distant metastasis or family history; 3) no neoadjuvant therapy; 4) lymphoma, multiple tumors, stump carcinoma and intraepithelial carcinoma Cases with neoplastic changes were excluded; 5) At least three gastrointestinal pathology experts evaluated the tumor stage and lymph node metastasis status. 12 pairs of gastric cancer with lymph node metastasis and 11 pairs of gastric cancer with non-lymph node metastasis, a total of 46 samples, all fresh and frozen tissue samples, the clinical information of the samples is shown in Table 1.1:
[0047]
[0048]
[0049] 2. Experiment, data analysis and biomarker screening:
[0050] The above samples are used to build a library of fresh frozen tissue or FFPE samples. The process and steps are as follows:
[0051] 1. Tissue DNA extraction ...
Embodiment 2
[0127]Using the 12 pairs of gastric cancer lymph node metastasis cancer and adjacent cancer and 11 pairs of gastric cancer non-lymph node metastasis cancer and adjacent cancer described in Example 1, a total of 46 samples, using the 60 biomarkers (markers) screened in Example 1 to establish Divide these 48 samples into 100 times according to the ratio of 7:3, use Random Forest (Random Forest) to model, use Random Forest (Random Forest) to model in the train set in each segmentation, use this model The risk score of each sample is calculated in the test set, and the risk score is compared with the standard diagnosis to obtain the discrimination sensitivity, specificity, AUC, NPV, and PPV of the combination of methylated regions. Specific allocation: Randomly select markers based on 4marker (chr12:42873140, chr8:81789921, chr8:81811441 and chr8:16884489), respectively 4marker (chr12:42873140, chr8:81789921, chr8:81811441, chr8:16884489) 10marker(chr12:42873140、chr8:81789921、chr8...
Embodiment 3
[0130]On the basis of the 60 biomarkers (markers) screened in Example 1, randomly select biomarkers from the 1366 biomarkers (markers) and 1306 biomarkers (markers) except 60 marker and 60 biomarkers (marker) combined into 100 biomarkers (marker) (chr1:2980508, chr1:9054472, chr1:53387866, chr10:15032997, chr10:129535510, chr11:460694, chr12:6729752 、chr12:42873140、chr12:52644274、chr12:107713297、chr13:50707693、chr13:113765062、chr14:48145305、chr15:21968071、chr16:22825741、chr16:31487303、chr16:55358704、chr16:86542314、chr17:6617301、chr19 :4054950、chr19:12876993、chr19:40421554、chr19:46388022、chr19:46388262、chr19:46388316、chr19:46388329、chr19:55593563、chr19:57352008、chr2:99439396、chr2:115420396、chr2:210636540、chr20:43726625 、chr20:54579248、chr20:57429986、chr20:59827184、chr20:59828473、chr20:61472147、chr20:61638536、chr21:22370447、chr4:20254852、chr5:2749179、chr5:140799056、chr5:163754234、chr6:85473377、chr6 :118228600、chr7:330023、chr7:47092638、chr7:56242574、chr7:71801447、chr7:98971536、c...
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