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Application of miR-34c in preparation of medicaments or health-care products for treating or preventing liver cancers

1. The technology of mir-34c and health products, applied in the application field of primary liver cancer, can solve the problems that there is no research on the regulation mechanism of miR-34 liver cancer genes, and it is difficult to determine

Inactive Publication Date: 2011-08-10
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The current research on the miR-34 family is mainly focused on miR-34a, and it is still difficult to determine whether the conclusions derived from miR-34a are also applicable to miR-34b / c
There are few studies on miR-34 in liver cancer, and there is no research on the gene regulation mechanism of miR-34 on HBV infection and its related liver cancer

Method used

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  • Application of miR-34c in preparation of medicaments or health-care products for treating or preventing liver cancers
  • Application of miR-34c in preparation of medicaments or health-care products for treating or preventing liver cancers
  • Application of miR-34c in preparation of medicaments or health-care products for treating or preventing liver cancers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Gene chip detection of expression of miRNAs and mRNAs in HBV transgenic Balb / c mouse liver cancer tissue and normal mouse liver tissue

[0020] According to the instructions of Trizol reagent, extract the total RNA of cells: take 6, 12, and 20-month-old HBV transgenic Balb / c mouse liver tissue and normal Balb / c mouse liver tissue of bean size, add 1ml Trizol extract, grind, Pipette and mix well, collect the suspension into DEPC-treated Ep tubes, and incubate the samples at 15°C to 30°C for 5 minutes to fully separate the nucleic acid and protein. Add 0.2ml chloroform, shake vigorously for 15s, incubate at 15°C-30°C for 2-3min, centrifuge at 12000rpm at 4°C for 15min, take the RNA supernatant, transfer it to a new Ep tube, add 0.5ml isopropanol, mix upside down Evenly, incubate at 15°C-30°C for 10min, centrifuge at 12000rpm at 4°C for 10min, discard the supernatant, add 1ml 75% DEPC-ethanol, rinse the precipitate, centrifuge at 7500rpm, 2°C-8°C for 5min. Disc...

Embodiment 2

[0022] Example 2: Prediction of miR-34c target genes

[0023] In conjunction with the result of embodiment 1 and bioinformatics method, utilize online database (TargetScan: http: / / www.targetscan.org / ; Pictar: http: / / pictar.bio.nyu.edu / ; miRanda: http: / / microrna.sanger.ac.uk / ) predicts the target gene related to miR-34c, tentatively confirms that the target gene of miR-34c is TGIF 2 .

Embodiment 3

[0024]Example 3: Real-time quantitative RT-PCR detection of miR-34c and TGIF in liver cancer cell lines 2 The expression level

[0025] The total RNA of liver cancer cell lines HepG2 and HepG2.2.15 cells were extracted by Trizol method. Wash the cells twice with PBS, add 1ml Trizol to each bottle, pipette to mix, and incubate at room temperature for 5min; transfer the cell solution into a DEPC-treated 1.5ml centrifuge tube, add 0.2ml chloroform to each tube, shake vigorously for 15s to mix, Stand on ice for 3-5 minutes to separate layers; centrifuge at 12,000 rpm for 15 minutes at 4°C; carefully absorb the upper layer liquid, transfer it to another DEPC-treated 1.5ml centrifuge tube, add an equal volume (0.5ml) that has been pre-prepared at 4°C Cold isopropanol, upside down and gently mix for 15-30s, let stand on ice (room temperature) for 5-10min; 4℃, 12000rpm, centrifuge for 15min; discard the supernatant, add 1ml 75% ethanol, gently wash the precipitate 2 centrifuge at 75...

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Abstract

The invention discloses application of non-coded small RNA gene miR-34c in diagnosis and treatment of primary liver cancers. The miR-34c can effectively inhibit hepatitis B virus (HBV) related liver cancers. Discovered by researches, the miR-34c has remarkable low expression in HBV transgenic mice liver cancer tissues and HBV infected liver cancer cell strains; and after expression vectors of the miR-34c are transfected to liver cancer cells HepG2.2.15, the miR-34c can remarkably inhibit replication and expression of HBV DNA, inhibit proliferation of liver cancer cells and promote apoptosis of liver cancer cells, and the researches show that the miR-34c can be helpful for early diagnosis and prognosis and treatment of the HBV related liver cancers, so the miR-34c can be used for preparing medicaments or health-care products for treating or preventing the liver cancers, particularly the primary liver cancers caused by HBV infection.

Description

technical field [0001] The invention relates to non-coding small RNA gene miR-34c and its target gene TGIF 2 especially related to its application in early diagnosis, prevention and treatment of primary liver cancer, especially primary liver cancer caused by HBV infection. Background technique [0002] Primary hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. Its morbidity and mortality are high, and it seriously endangers people's health. Fetal protein (AFP) is the most commonly used detection method, but sometimes missed and misdiagnosed, and there is no satisfactory treatment. [0003] Hepatitis B virus (HBV) infection is considered to be the main cause of HCC. 60% of HCC cases worldwide and 70% of cases in endemic areas are thought to be caused by HBV. The mechanism of HBV-induced primary liver cancer is extremely complex. [0004] Studies have shown that in the process of some virus infection, the virus can regulate the gene exp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/7105A61P35/00
Inventor 刘玉刚王燕王春梅范春光孙汶生
Owner SHANDONG UNIV