CN115300505A

CN115300505A - Application of FB23 in preparation of medicine for treating liver cancer

Info

Publication number: CN115300505A
Application number: CN202210909595.7A
Authority: CN
Inventors: 宋关斌; 陈哲; 蒋灵丽; 罗庆
Original assignee: Chongqing University
Current assignee: Chongqing University
Priority date: 2022-07-29
Filing date: 2022-07-29
Publication date: 2022-11-08

Abstract

The invention relates to an application of FB23 in a medicine for treating liver cancer, and the FB23 can inhibit the proliferation and migration behaviors of liver cancer cell lines MHCC97H, HCCLM and HepG2 and has the effect of promoting the apoptosis of MHCC97H and HepG 2. The FB23 has good application prospect in treatment of liver cancer.

Description

Application of FB23 in preparation of medicine for treating liver cancer

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of FB23 in preparation of a medicine for treating liver cancer.

Background

The primary liver cancer is one of the most common clinical malignant tumors, and the morbidity and the mortality of the primary liver cancer are in the front of the malignant tumors. According to the latest data of the international agency for research on cancer (IARC) in world health, liver cancer is the sixth most frequently diagnosed cancer and the third leading cause of death worldwide in 2020, and hepatocellular carcinoma (HCC) accounts for 75% -85% of primary liver cancer. With the awareness of the severity and risk of HCC, drug and non-drug treatment have made great progress in the HCC treatment field. Non-drug therapies include radical resection, liver transplantation, transcatheter arterial chemoembolization, percutaneous ablation, and immunotherapy. The drug therapy mainly comprises small molecule targeted drug therapy, such as sorafenib, lenvatinib and the like. Despite the diversity of HCC treatment, the overall survival of patients remains far from satisfactory. Therefore, it is of great importance to develop more effective treatments for patients.

m6A modification is a methylation modification of RNA that is widely found in many species, including mammals. The m6A modification is enriched in the stop codon, 3' untranslated region and near the coding region, which are critical for regulating mRNA stability, splicing, transport, translation, primary microRNA processing and protein-RNA interactions. The precise regulation of m6A modifications on RNA plays an important role in a variety of biological and pathological processes.

The m6A modification is mediated by methylases and demethylases, FTO belonging to the Fe family ²⁺ And the 2-oxoglutarate-dependent AlkB dioxygenase family, which predominantly catalyze m6A demethylation. The effects of FTO demethylation dysfunction are associated with human disease, particularly in cancer. Studies have demonstrated that FTO small molecule inhibitors can transiently interfere with mRNA methylation.

FB23 is a synthetic compound, an inhibitor of FTO, having a molecular formula of C ₁₈ H ₁₄ Cl ₂ N ₂ O ₃ The molecular weight of 377.22 has the following structure.

The research shows that FB23 can inhibit the proliferation of human acute myeloid cell line and promote the apoptosis of cells.

In the research on the influence of FB23 on liver cancer cell lines, the applicant finds that FB23 can inhibit the proliferation and migration of liver cancer cell lines and promote the apoptosis of cell lines, and the result is not reported at home and abroad at present.

Disclosure of Invention

The invention aims to provide application of FB23 in preparation of a medicament for treating liver cancer, wherein the FB23 can effectively inhibit proliferation and migration of a liver cancer cell line and promote apoptosis. Under the premise of lacking FTO inhibitor at present, FB23 has good application prospect.

The technical scheme of the invention is as follows:

FB23 can be used for preparing medicine for treating hepatocarcinoma.

The drug is an inhibitor.

The FB23 can inhibit the proliferation of MHCC97H, HCCLM3 and HepG2 cells of liver cancer cells.

The FB23 promotes the apoptosis of MHCC97H and HepG2 cells of liver cancer cells.

The FB23 inhibits the migration of MHCC97H, HCCLM3 and HepG2 cells of liver cancer cells.

The applicant shows that the drug (compound FB 23) can inhibit the proliferation and migration behaviors of the liver cancer cell lines MHCC97H, HCCLM and HepG2 and has the effect of promoting the apoptosis of MHCC97H and HepG2 through the experiments of the influence of FB23 on the proliferation of the liver cancer cell lines, the influence of FB23 on the apoptosis rate of the liver cancer cell lines and the influence of FB23 on the migration of the liver cancer cell lines, so that the drug can play a role in inhibiting cancer in HCC by inhibiting the proliferation and migration of the liver cancer cell and promoting the apoptosis, and provides a new treatment strategy for HCC.

The medicine can be used for treating liver cancer and has good application prospect.

Drawings

FIG. 1 is the effect of FB23 on hepatocarcinoma cell proliferation;

FIG. 2 is the effect of FB23 on apoptosis of hepatoma cells;

FIG. 3 shows the effect of FB23 on liver cancer cell migration.

Detailed Description

FB23 used in the experiments of this example was purchased from Selleck;

human hepatoma cell lines MHCC97H and HCCLM3 were purchased from Shanghai Oriental hepatobiliary Hospital, and HepG2 was purchased from American type culture Bank (ATCC, manassas, WV, USA);

Annexin-PI apoptosis kit purchased from BD company;

transwell chambers were purchased from Corning corporation;

DMEM medium and newborn bovine serum used in the experiments of this example were purchased from Gibco;

other reagents used in the experiments of this example were commercially available analytical grade reagents.

Example 1: effect of FB23 on the proliferation of liver cancer cell lines

1.1 Experimental methods

The initial number of inoculations in each 24-well plate was 1X 10 ⁴ MHCC97H, HCCLM3 and HepG2 cell lines per well, then added exogenously at concentrations of 0.1% DMSO, 5. Mu.M FB23 and 10. Mu.M FB23, respectively. The number of cells was counted every 24 hours, and counting was stopped after day 4 and a proliferation curve was plotted.

1.2 results of the experiment

The cell proliferation curve results show that FB23 can significantly inhibit the proliferation of MHCC97H, HCCLM3 and HepG2 cells (see fig. 1).

And (4) conclusion: FB23 can inhibit proliferation of hepatocarcinoma cell.

Example 2: effect of FB23 on the apoptosis Rate of liver cancer cell lines

2.1 Experimental methods

MHCC97H and HepG2 cell lines were inoculated, respectively, into 12-well plates, and after 24 hours of culture, treated with 0.1% DMSO and 10. Mu. MFB23, respectively, for 48 hours. After the cells are collected, annexin V and PI staining solution is prepared, namely under a 100 mu L Binding buffer system, 2.5 mu L V-FITC and 2.5 mu L PI are added, 100 mu L staining solution is added into each sample, and after 1 hour of normal temperature staining, the samples are analyzed and detected by a flow cytometer.

2.2 results of the experiment

Apoptosis analysis showed that 10 μ MFB23 was effective in promoting apoptosis in MHCC97H and HepG2 cells with Annexin V positive ratios increasing from 6.48% and 9.05% to 25.88% and 26.72%, respectively (see fig. 2).

And (4) conclusion: FB23 can increase apoptosis of hepatocarcinoma cell.

Example 3: effect of FB23 on migration of liver cancer cell lines

3.1 Experimental methods

The number of inoculations in each upper chamber of the Transwell chamber was 2X 10 ⁴ The MHCC97H, HCCLM3 and HepG2 cell lines of (1) were prepared by adding 600. Mu.l of 10% FBS high-sugar medium to the upper chamber and then exogenously adding DMSO at a concentration of 0.1% to the lower chamber and 5. Mu.M FB23 and 10. Mu.M FB23, respectively. After 24 hours of action, the filter was fixed with 4% paraformaldehyde for 30min after swabbing the non-migrated cells from the upper layer of the filter with a cotton swab. After fixation was completed, the plate was stained with 0.1% crystal violet for 30min. Washing off excessive crystal violet dye solution with PBS, air drying, and developingAnd (3) randomly selecting 5 different areas in the upper, lower, left and right directions for photographing in each small chamber under the micro-mirror, finally counting the number of migrated cells, and calculating the number of migrated cells in each group of samples.

3.2 results of the experiment

Migration results showed that FB23 can inhibit MHCC97H, HCCLM3 and HepG2 cell migration (see fig. 3).

And (4) conclusion: FB23 can block the migration of liver cancer cells.

Claims

An application of FB23 in preparing the medicine for treating liver cancer.
2. Use according to claim 1, characterized in that: the drug is an FTO inhibitor.
3. Use according to claim 1, characterized in that: the FB23 can inhibit the proliferation of MHCC97H, HCCLM3 and HepG2 cells of liver cancer cells.
4. Use according to claim 1, characterized in that: the FB23 promotes the apoptosis of MHCC97H and HepG2 cells of liver cancer cells.
5. Use according to claim 1, characterized in that: the FB23 can inhibit the cell migration of MHCC97H, HCCLM3 and HepG2 of liver cancer cells.