miRNAs for the diagnosis and treatment of hepatocellular carcinoma

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

Active Publication Date: 2020-11-17
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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  • miRNAs for the diagnosis and treatment of hepatocellular carcinoma
  • miRNAs for the diagnosis and treatment of hepatocellular carcinoma
  • miRNAs for the diagnosis and treatment of 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, biotechnology, catalog number B532451-0020,

[0054] miRNA fluorescence quantitative PCR kit, Shenggong, catalog number B532461-0002,

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

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

[0057] 2. Experimental samples

[0058] Collect 32 cases of histopathologically diagnosed cancer tissues of patients with primary liver cancer and their corresponding> The 3cm para-cancer tissue samples, all samples were put into cryopreservation tubes within 30 minutes of surgical excision, put into liquid nitrogen tank quickly, and transferred to the laboratory -80℃ ultra-low temperature refrigerator.

[0059] Exclusion criteria: Having undergone anti-tumor treatment before surgery, suffering from othe...

Embodiment 2

[0093] Example 2 Verification of 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 adjacent tissues were collected from the TCGA database, and the expression level of miR-4772-3p in liver cancer tissues and adjacent tissues was analyzed.

[0096] 2. ROC curve analysis

[0097] Use the pROC package in R language to analyze receiver operating characteristics of miR-4772-3p, calculate the binomial accurate confidence space, and draw the ROC curve.

[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 higher area under the curve, indicating that the use of miR-4772-3p for the diagnosis of liver cancer has higher sensitivity and specificity, and has better Diagnostic efficiency.

Embodiment 3

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

[0101] 1. Cell culture

[0102] Cultivation of human normal liver cell line L-02, low invasion and metastasis liver cancer cell line HepG2, and high invasion and metastasis liver cancer cell line Huh7. Cells are routinely cultured in 10% FBS, 100 units / ml penicillin and 100ug / ml chain In the 1640 cell culture medium of mycin, at 37℃, 5% CO 2 And cultivate under saturated humidity conditions. The cells were changed every 2d and passaged at 1:3.

[0103] 2. Extraction of cellular RNA

[0104] The TRIzol method was used to extract total cellular RNA.

[0105] Collect the cells in the log phase, add 1ml TRIzol, and mix well; use a 1ml syringe to repeatedly rupture the cells and cut the DNA, leave it at room temperature for 5 minutes, and the remaining 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 ...

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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 miRNAs for diagnosis and treatment of liver cancer. Background technique [0002] Primary liver cancer is the world's fourth leading cause of cancer deaths and has the fastest increase in incidence among all cancers. About 800,000 people die from primary liver cancer each 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 hepatocytes Hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (Intrahepaticcholangiocarcinoma, ICC), hepatocellular-cholangiocyte mixed carcinoma, among which HCC has the highest incidence, accounting for approximately 90% of primary liver cancer (Wakizaka K, Yokoo H ,K...

Claims

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Application Information

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Patent Type & Authority Patents(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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