Diagnosis product and curing composition for hepatocellular carcinoma
A composition and technology for liver cancer, applied in the field of biomedicine, can solve the problems of lack of sensitivity and specificity in early diagnosis of liver cancer
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Embodiment 1
[0052] Example 1 QPCR detection of miR-6761-5p expression in hepatocellular carcinoma
[0053] 1. Experimental materials
[0054] 1. Experimental reagents and instruments
[0055] miRNA first-strand cDNA synthesis, Sangon, Cat. No. B532451-0020,
[0056] miRNA Fluorescent Quantitative PCR Kit, Sanko, Cat. No. B532461-0002,
[0057] NanoVue Plus, BIOCHROM LTD, model 28956057,
[0058] Fluorescent quantitative PCR instrument, Applied Biosystems, model ABI7300
[0059] 2. Experimental samples
[0060] The cancer tissues and the corresponding paracancerous tissue samples > 3cm were collected from 32 cases of primary liver cancer patients diagnosed by histopathology. All samples were put into cryopreservation tubes within 30 minutes after surgical resection, and quickly put into liquid nitrogen tanks. , and transferred to the laboratory -80°C ultra-low temperature freezer.
[0061] Exclusion criteria: patients who had undergone anti-tumor treatment before operation, and had a...
Embodiment 2
[0094] Example 2 Verifying the diagnostic efficacy of biomarkers
[0095] 1. Data collection
[0096] The miRNA expression profile data of 342 cases of liver cancer tissues and 50 cases of paracancerous tissues were collected from the TCGA database, and the expression levels of miR-6761-5p in liver cancer tissues and paracancerous tissues were analyzed.
[0097] 2. ROC curve analysis
[0098] The receiver operating characteristics of miR-6761-5p were analyzed using the pROC package in R language, the binomial exact confidence space was calculated, and the ROC curve was drawn.
[0099] 3. Results
[0100] The ROC analysis results of miR-6761-5p are as follows figure 2 As shown, the AUC value of miR-6761-5p as the test result variable is 0.744, which has a high area under the curve, indicating that the use of miR-6761-5p in the diagnosis of liver cancer has high sensitivity and specificity, and has a good diagnostic performance.
Embodiment 3
[0101] Example 3 Detection of the expression level of miRNA in liver cancer cells
[0102] 1. Cell culture
[0103] Culture human normal liver cell line L-02, liver cancer cell line HepG2 with low invasion and metastasis ability, and liver cancer cell line Huh7 with high invasion and metastasis ability. Mycin 1640 cell culture medium at 37°C, 5% CO 2 and cultured under conditions of saturated humidity. The cells were replaced every 2 days and passaged at a ratio of 1:3.
[0104] 2. Extraction of cellular RNA
[0105] Total cellular RNA was extracted using the TRIzol method.
[0106] Collect the cells in the logarithmic phase, add 1ml TRIzol, and mix well; draw repeatedly with a 1ml syringe to rupture the cells and cut the DNA, let stand at room temperature for 5min, and the rest of the steps are the same as in Example 1.
[0107] 3. Reverse transcription and fluorescence quantitative detection
[0108] The experimental process is the same as in Example 1.
[0109] 4. St...
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