miRNA (micro ribonucleic acid) for diagnosis and treatment on hepatocellular carcinoma

A technology of hepatocellular carcinoma and reagents, applied in the field of miRNA

Active Publication Date: 2020-05-19
THE THIRD HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although small-molecule targeted therapy drugs can improve the overall survival of HCC patients, however, only a slight improvement in the survival of HCC patients has been observed with these treatments due to increasing drug resistance in patients

Method used

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  • miRNA (micro ribonucleic acid) for diagnosis and treatment on hepatocellular carcinoma
  • miRNA (micro ribonucleic acid) for diagnosis and treatment on hepatocellular carcinoma
  • miRNA (micro ribonucleic acid) for diagnosis and treatment on hepatocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 QPCR detection of miR-4772-3p expression in hepatocellular carcinoma

[0051] 1. Experimental materials

[0052] 1. Experimental reagents and instruments

[0053] miRNA first-strand cDNA synthesis, Sangon, Cat. No. B532451-0020,

[0054] miRNA Fluorescent Quantitative PCR Kit, Sanko, Cat. No. B532461-0002,

[0055] NanoVue Plus, BIOCHROM LTD, model 28956057,

[0056] Fluorescent quantitative PCR instrument, Applied Biosystems, model ABI7300

[0057] 2. Experimental samples

[0058] 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.

[0059] Exclusion criteria: patients who had undergone anti-tumor treatment before operation, and had a...

Embodiment 2

[0093] Example 2 Verifying the diagnostic efficacy of biomarkers

[0094] 1. Data collection

[0095] 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-4772-3p in liver cancer tissues and paracancerous tissues were analyzed.

[0096] 2. ROC curve analysis

[0097] The receiver operating characteristics of miR-4772-3p were analyzed using the pROC package in R language, the binomial exact confidence space was calculated, and the ROC curve was drawn.

[0098] 3. Results

[0099] The ROC analysis results of miR-4772-3p are as follows figure 2 As shown, the AUC value of miR-4772-3p as the test result variable is 0.746, which has a high area under the curve, indicating that the use of miR-4772-3p in the diagnosis of liver cancer has high sensitivity and specificity, and has a good diagnostic performance.

Embodiment 3

[0100] Example 3 Detection of the expression level of miRNA in liver cancer cells

[0101] 1. Cell culture

[0102] 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.

[0103] 2. Extraction of cellular RNA

[0104] Total cellular RNA was extracted using the TRIzol method.

[0105] 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.

[0106] 3. Reverse transcription and fluorescence quantitative detection

[0107] The experimental process is the same as in Example 1.

[0108] 4. St...

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Abstract

The invention discloses a miRNA (micro ribonucleic acid) for diagnosis and treatment on hepatocellular carcinoma. The invention finds for a first time that expression of miR-4772-3p is decreased in liver cancer, and further verifies that miR-4772-3p has high diagnosis efficiency in liver cancer. The invention further discloses that miR-4772-3p plays significant roles in proliferation, migration and invasion of hepatoma carcinoma cells, and reminds that miR-4772-3p can be applied to treatment on liver cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to miRNA for the diagnosis and treatment of liver cancer. Background technique [0002] Primary liver cancer is the fourth leading cause of cancer-related deaths worldwide and has the fastest-growing incidence among all cancers. About 800,000 people die from primary liver cancer every year, and the mortality rate is still increasing at a rate of 3% per year (Siegel RL, MillerKD. Cancer statistics, 2019[J]. 2019; 69(1): 7-34; Bertuccio P, Turati F, Carioli G, et al.Global trends and predictions in hepatocellμlar carcinoma mortality[J].Journal of hepatology.2017;67(2):302-9.), the histological types of primary liver cancer include hepatocellular Hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC), mixed hepatocellular-cholangiocarcinoma, among which HCC has the highest incidence, accounting for about 90% of primary liver cancer (Wakizaka K, Yokoo H , Kamiyama T, et al. Cli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K31/7105A61P35/00A61P1/16
CPCA61K31/7105A61K45/00A61P1/16A61P35/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 窦剑赵鑫张颖刘文鹏曾强王洋刘宝旺崔自强滕亮张军红曹经琳高庆军
Owner THE THIRD HOSPITAL OF HEBEI MEDICAL UNIV
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