Gene marker of predicting primary hepatic carcinoma metastatic potential
A technology of sulfur transfer and gene chip, applied in the field of medicine
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preparation example Construction
[0033] The preparation of the gene chip of the present invention can adopt the conventional manufacturing method of biochip. For example, if the solid phase carrier is a modified glass slide or a silicon wafer, and the 5' end of the probe contains amino-modified poly dT strings, the oligonucleotide probe can be formulated into a solution, and then the spotting instrument will spot it on The gene chip of the present invention can be obtained by modifying glass slides or silicon slices, arranging them into a predetermined sequence or array, and then fixing them by placing them overnight. If the oligonucleotide probe does not contain amino modification, its preparation method can also refer to: "Gene Diagnosis Technology-Non-radioactive Operation Manual" edited by Wang Shenwu; J.L.erisi, V.R.Iyer, P.O.BROWN.Exploring the metabolic and genetic control of gene expression on a genomic scale. Science, 1997; 278: 680 and Ma Liren, edited by Jiang Zhonghua. Biochip. Beijing: Chemical I...
Embodiment 1
[0050] Embodiment 1, the preparation of RNA sample
[0051] Tissue sample:
[0052] 49 cases of cancer tissue (including 28 samples with metastases and 21 samples without metastases) were obtained from surgical resection specimens of patients with primary liver cancer. Normal livers were obtained from accidental deaths. All the above-mentioned specimens were obtained with the approval of the ethics committee of the WHO partner organization authorized by the Shanghai Municipal Government. The clinical data of tissue samples include: gender, age, tumor size, pathological grade (Edmonson), liver cirrhosis, metastasis, recurrence, HBV and HCV infection, etc. All patients did not receive chemotherapy before operation, and the postoperative follow-up was up to 60 months.
[0053] Gene chip:
[0054] Whole-genome expression profile chip, using the crystal core of Capital Bio Co., Ltd. (http: / / www.capitalbio.com) Human whole genome oligonucleotide microarray chip V1.0 (dual-chan...
Embodiment 2
[0060] Example 2, screening genes with significant differences
[0061] 28 cases of cancer samples that have metastasized and normal liver samples (10 cases mixed), 21 cases of non-metastatic cancer samples and normal liver samples (10 cases mixed) were labeled with two fluorescent dyes, cy3 and cy5, and expressed in the whole genome Competitive hybridization of probes on a spectrum chip. Chip core after hybridization LuxScan TM The 10K microarray chip scanner scans to obtain the result image, and then quantifies the hybridization result through the LuxScan 3.0 general microarray image analysis software that comes with it. Thus, the expression profiles of 28 cases with transfer samples to normal samples (with transfer / normal, Metastasis / Normal) mRNA chips and 21 cases without transfer samples to normal samples (without transfer / normal, Non-Metastasis / Normal) mRNA chips were obtained data.
[0062] The microarray results of the above 28 cases with metastasis / normal and 10...
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