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Application of SHMT2 inhibitor in preparation of drugs for inhibiting growth and metastasis of bladder cancer cells

1. The technology of SHMT2 and inhibitors, which is applied in the field of medicine, can solve the problems that the therapeutic effect of bladder cancer has not been reported yet.

Active Publication Date: 2021-05-07
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] There is no report on the therapeutic effect of SHIN1 and Crolibulin on bladder cancer

Method used

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  • Application of SHMT2 inhibitor in preparation of drugs for inhibiting growth and metastasis of bladder cancer cells
  • Application of SHMT2 inhibitor in preparation of drugs for inhibiting growth and metastasis of bladder cancer cells
  • Application of SHMT2 inhibitor in preparation of drugs for inhibiting growth and metastasis of bladder cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Culture, expansion and passage of bladder cancer cells;

[0051]Human bladder cancer cell lines BIU-87, EJ, and T24 were selected and grown in RPMI-1640 medium. Human kidney epithelial cell line 293T cells were grown in DMEM medium. Both the RPMI-1640 medium and the DMEM medium contain penicillin (100 U / ml) and streptomycin (100 mg / ml) and 10% fetal bovine serum. The above cells were incubated at 37°C in a humidified atmosphere of 5% CO2.

Embodiment 2

[0053] Cell Proliferation Assay

[0054] Bladder cancer cell lines BIU87, T24, EJ were routinely cultured in RPMI-1640 medium containing 10% fetal bovine serum, and human kidney epithelial cell line 293T was routinely cultured in DMEM medium containing 10% fetal bovine serum. Digest BIU87, T24, EJ, 293T cells, wash once with 5ml PBS, and count accurately. Plate 5000 cells / 100 μl / well in a 96-well plate and culture overnight. After the cells adhered to the wall the next day, they were treated with different concentrations of Crolibulin and SHIN1 for 24 hours and 48 hours. The concentrations of Crolibulin included 0 μM, 1 μM, 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, and the concentrations of SHIN1 included 0 nM and 1 nM , 10nM, 20nM, 30nM, 40nM, 50nM, 60nM, 70nM, the concentration of RBC8 includes 0μM, 5μM, 10μM, 15μM, the concentration of UCPH-101 includes 0.5μM, 1μM, 1.5μM, 2μM. At each time point, remove the original medium, add 100 μl of CCK8-containing medium, inc...

Embodiment 3

[0058] Apoptosis assay

[0059] BIU87 cells were divided into 4 × 10 per well 5 The cells were seeded in two 6-well plates, and after adhering to the wall the next day, fresh medium containing 2 μl DMSO was added to well 3, fresh medium containing 11 μM Crolibulin inhibitor was added to well 3, and fresh medium containing 29 nM SHIN1 inhibitor was added to well 3 fresh medium for 24 hours. After 24 hours, suck out the cell culture medium into a suitable centrifuge tube, wash the adherent cells once with pre-cooled PBS, and add an appropriate amount of trypsin cell digestion solution to digest the cells. Incubate at room temperature until the adherent cells can be blown down by gentle pipetting, then aspirate the trypsin cell digestion solution. Overdigestion with trypsin should be avoided. Add the collected cell culture solution, mix slightly, transfer to a centrifuge tube, and centrifuge at 1000rpm for 5 minutes to pellet the cells. Dilute the Binding Buffer 1:10 with dei...

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Abstract

The invention provides an application of an SHMT2 inhibitor and an analogue thereof in preparation of drugs for inhibiting the growth and metastasis of tumor cells. The SHMT2 inhibitor and the analogue thereof are SHIN1 and Crolibulin. In-vitro research shows that the SHMT2 inhibitor and the structural analogues thereof (SHIN1 and Crolibulin) can effectively inhibit bladder cancer cell proliferation, promote bladder cancer cell apoptosis, inhibit bladder cancer cell cycle, inhibit bladder cancer cell migration, reduce bladder cancer cell clone formation ability and inhibit stemness maintenance of bladder cancer stem cells, so that the SHMT2 inhibitor and the analogue thereof can be used for preventing and / or treating bladder cancer and can be used as the safe and effective treatment drugs for treating the bladder cancer.

Description

technical field [0001] The invention belongs to the field of medicine, and specifically relates to the application of SHMT2 inhibitors and analogues thereof in the preparation of drugs for inhibiting tumor cell growth and metastasis, in particular to the application of the same in the preparation of drugs for inhibiting bladder cancer cell growth and metastasis. Background technique [0002] Bladder cancer refers to malignant tumors that occur on the bladder mucosa, usually bladder urothelial cell carcinoma. In my country, bladder cancer is the most common malignant tumor in the urinary system. In the United States, bladder cancer is also the second most common malignant tumor in the urinary system after prostate cancer. In general, the incidence of bladder cancer increases with age and is much more common in men than in women. Due to the development of industrialization and population aging, the incidence of bladder cancer in my country is still severe. Statistics show th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P35/00
CPCA61K45/00A61P35/00
Inventor 温廷益刘喆张芸邓爱华刘树文
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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