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Application of reagent for reducing expression or activity of LC-3B in preparing of drugs for enhancing anti-hepatocarcinoma activity of lycorine

An anti-liver cancer, lycorine technology, applied in anti-tumor drugs, drug combinations, organic active ingredients and other directions, can solve problems such as unclear molecular mechanism

Active Publication Date: 2019-02-05
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its precise molecular mechanism is still unclear

Method used

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  • Application of reagent for reducing expression or activity of LC-3B in preparing of drugs for enhancing anti-hepatocarcinoma activity of lycorine
  • Application of reagent for reducing expression or activity of LC-3B in preparing of drugs for enhancing anti-hepatocarcinoma activity of lycorine
  • Application of reagent for reducing expression or activity of LC-3B in preparing of drugs for enhancing anti-hepatocarcinoma activity of lycorine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1. Autophagy inhibitors enhance the anti-liver cancer activity of lycorine (the combination of autophagy inhibitors and lycorine enhances the apoptosis of liver cancer cells)

[0064] 1. Lycorine inhibits the growth of hepatocellular carcinoma

[0065] The cell viability was determined by MTT assay, and the growth inhibitory effect of lycorine on three typical human hepatocellular carcinoma cell lines (HepG2, SMMC-7721, HuH-7) and the non-tumorigenic human liver cell line (ChangLiver ). HepG2, SMMC-7721, HuH-7 and Chang Liver were cultured for 48 hours with the medium containing lycorine at 0, 10, 20, 30, 40 and 50 μM respectively, and the cell viability was determined by MTT method. The results showed that 10, 20 , 30, 40, and 50 μM lycorine were treated for 48 hours, and the viability of liver cancer cells was significantly reduced. At the same time, lycorine did not affect the cell viability of normal liver cells Chang Liver at the same dose ( figure 1 Midd...

Embodiment 2

[0084] Example 2. siRNA targeting LC-3B enhances the anti-liver cancer activity of lycorine, and siRNA targeting LC-3B and lycorine have a synergistic effect in anti-liver cancer

[0085] NCBI Reference Sequence of Human Microtubule-Associated Protein 1 Light Chain 3β (Homo sapiens microtubule associated protein 1 light chain 3beta, MAP1LC3B, LC-3B for short) gene: NM_022818.4 (Update Date: 10-JUL-2017).

[0086] This experiment selects random double-stranded RNA as negative control siRNA, and the name of this random double-stranded RNA is siNC, and the nucleotide sequence from the 5' end to the 3' end of a strand of siNC is (5'-uucuccgaacgugucacguTT-3'), The nucleotide sequence from the 3' end to the 5' end of the other chain of siNC is (3'-TTaagaggcuugcacagugca-5').

[0087] In this experiment, two siRNAs targeting human LC-3B were selected, named LC-3B-1 and LC-3B-2 respectively, the nucleotide sequence from the 5' end to the 3' end of a chain of LC-3B-1 For (5'-gcucuucuag...

Embodiment 3

[0099] Example 3, Akt inhibitor enhances the anti-liver cancer activity of lycorine (the combination of Akt inhibitor and lycorine enhances the apoptosis of liver cancer cells)

[0100] 1. Lycorine promotes apoptosis and autophagy of liver cancer cells through TCRP1 / Akt / mTOR signaling pathway

[0101] The experiment was carried out with reference to the corresponding experimental method in Example 1.

[0102] Tongue cancer resistance-associated protein 1 (TCRP1) was reported to promote tumorigenesis in radiation-resistant oral squamous cell carcinoma by selectively activating PI3K / Akt and NF-KB pathways. Such as Figure 4 As shown in middle a, lycorine treatment significantly reduced the expression of TCRP1 protein level in hepatoma cells. However, lycorine treatment did not significantly alter the mRNA levels of TCRP1 ( Figure 4 In b), this indicates that the TCRP1 protein is largely transferred through proteasomal degradation. Subsequently, treatment with MG-132 (a prot...

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Abstract

The invention discloses application of a reagent for reducing expression or activity of LC-3B in preparing of drugs for enhancing anti-hepatocarcinoma activity of lycorine, and particularly provides application of a reagent for reducing expression or activity of LC-3B in preparing of products for enhancing anti-hepatocarcinoma activity or anti-hepatocarcinoma cell activity of lycorine. Experimental results show that the apoptotic rate of hepatocarcinoma cells treated by drug combination of the reagent for reducing the expression or activity of the LC-3B and the lycorine is greater than the sumof the apoptotic rate of the hepatocarcinoma cells treated alone by the reagent for reducing the expression or activity of the LC-3B and the apoptotic rate of the hepatocarcinoma cells treated aloneby the lycorine, indicating that the reagent for reducing the expression or activity of the LC-3B and the lycorine have a synergistic effect on anti-hepatocarcinoma, and the reagent for reducing the expression or activity of the LC-3B enhances the anti-hepatocarcinoma activity of the lycorine.

Description

technical field [0001] The present invention relates to the application of the reagent for reducing the expression or activity of LC-3B, in particular to the application of the reagent for reducing the expression or activity of LC-3B in the preparation of a drug for enhancing the anti-liver cancer activity of lycorine. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the most common aggressive tumors worldwide. Liver cancer is a highly lethal tumor with median survival remaining below 1 year after diagnosis. HCC is a heterogeneous disease originating from chronic liver diseases such as chronic inflammation and cirrhosis. So far, the treatment strategies for liver cancer mainly include resection, transplantation, local ablation and chemotherapy. Because the early symptoms of liver cancer are not obvious, most patients with liver cancer are diagnosed at advanced stage and lose the chance of surgery. Most patients who have undergone surgery in time will ...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K31/7105A61K48/00A61K31/437A61P35/00
CPCA61K45/06A61K31/437A61K31/7105A61P35/00A61K2300/00
Inventor 于海洋邱玉玲王涛刘二伟高秀梅陈璐吴红华尹双双
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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