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Application of glaucescent fissistigma root extract C21 steroid in preparation of medicine for promoting gastric cancer cell apoptosis

A technology for gastric cancer cells and cancer cells, which is applied in the field of application of C21 steroids of the extract of Radix vulgaris in the preparation of drugs for promoting gastric cancer cell apoptosis, which can solve the problems of unresearched anti-cancer effects, and achieve the enhancement of gastric cancer cell apoptosis. Effect

Inactive Publication Date: 2020-09-01
ZHEJIANG PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the potential efficacy of C21 steroids on autophagy and whether autophagy inhibition could contribute to the anticancer effects of C21 have not been investigated

Method used

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  • Application of glaucescent fissistigma root extract C21 steroid in preparation of medicine for promoting gastric cancer cell apoptosis
  • Application of glaucescent fissistigma root extract C21 steroid in preparation of medicine for promoting gastric cancer cell apoptosis
  • Application of glaucescent fissistigma root extract C21 steroid in preparation of medicine for promoting gastric cancer cell apoptosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Inhibitory effect of C21 steroids on the proliferation of gastric cancer cells

[0036] 50 μL of culture medium was added to each well of E-plate 96 (Roche) to obtain equilibrium; 2000 cells were then seeded in E-plate 96 . After 24 hours, add 40 μg / mL, 80 μg / mL, and 160 μg / mL of C21 steroids, respectively, and lock the E-plate 96 in the RTCA-MP device at 37 °C, CO 2 At a concentration of 5%, measured changes in electrical impedance, which directly reflect cell proliferation on the surface of the biocompatible microelectrode coating, were recorded, with automatic readings every two minutes.

[0037] 5000 cells were seeded in 96-well plates containing 200 μL RPIM1640 medium at 37 °C, 5% CO 2 and then treated with C21 steroids (0, 20 μg / mL, 40 μg / mL, 80 μg / mL, 160 μg / mL, 320 μg / mL) for 24 h; add 40 μL of MTS solution to each well and incubate for 2 h. Absorbance was measured at 490 nm with a microplate reader.

[0038] The result is as figure 1 Shown: RTCA growth curv...

Embodiment 2

[0041] Induction of cell cycle arrest and apoptosis in gastric cancer cells BGC-823 and AGS

[0042] Cells were seeded into 12-well plates incubated at 37°C for 24h, and then treated with C21 steroids (0, 80μg / mL, 160μg / mL) and incubated at 37°C for 24h, the cells were collected, and stained with 500μL staining buffer A and 5 μL of reagent B (Lianke Biotechnology) were treated, and then these cells were incubated in the dark at room temperature for 30 min, and the cell cycle was analyzed by flow cytometry.

[0043] The percentage of apoptotic cells was detected by Annexin-V-FITC and PI double staining, and then they were treated in the same manner as the cell cultures used in cell cycle analysis. Cells were collected, treated with 5 μL Annexin V-FITC and 10 μL PI, then incubated at room temperature in the dark for 5 min, and analyzed by flow cytometry for fluorescence-activated cells.

[0044] The result is as figure 2 Shown in a and b: the present invention adopts flow cyt...

Embodiment 3

[0048] Reactive Oxygen Species (ROS) Measurement

[0049] Cells were cultured for 24 h in the presence or absence of C21 steroid alone or with autophagy inhibitor CQ. After the cells were harvested and resuspended in 500 μL of PBS, they were stained with DCFH-DA (5 mm) for 30 min according to the manufacturer's instructions (Beyotime), and then the DCFH-DA signal was analyzed by flow cytometry.

[0050] To explore the biological role of C21 steroid-induced autophagy in cell survival or death, autophagy was blocked by autophagy inhibitor CQ, and gastric cancer cells were treated with C21 steroid alone or in combination with CQ for 24 hours. like Figure 4 Shown: Apoptosis experiments show that CQ promotes C21 steroid-induced apoptosis in gastric cancer cells. like Figure 4 As shown in a and 4b, the percentage of apoptosis in BGC-823 cells increased by 11.20% ± 1.69% (C21 steroid) and 17.70% ± 0.39% (C21 steroid + CQ) compared with the control group. The percentage of apopt...

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Abstract

The invention discloses application of a C21 steroid in preparation of a medicine for promoting cancer cell apoptosis. The C21 steroid inhibits cell proliferation by inhibiting autophagy to promote cell apoptosis. The invention further discloses application of the C21 steroid in preparation of a medicine for treating gastric cancer; the C21 steroid is a glaucescent fissistigma root extract; and the C21 steroid inhibits proliferation of gastric cancer cells by inhibiting autophagy and promoting apoptosis. The C21 steroid disclosed by the invention can obviously inhibit the growth of cancer cells and promote the cell apoptosis, and provides a theoretical basis for revealing a new mechanism of the anti-gastric cancer activity of the C21 steroid.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of the C21 steroid of the extract of Aegypti sinensis in the preparation of drugs for promoting the apoptosis of gastric cancer cells. Background technique [0002] Gastric cancer is the third most common cancer worldwide and one of the leading causes of cancer-related death. The lack of effective treatment methods leads to short survival period and poor quality of life of cancer patients. And at present, patients with gastric cancer are mainly treated by surgical resection and chemotherapy, but the therapeutic effect is limited. Therefore, there is an urgent need to develop new therapeutic options to improve patient outcomes. [0003] Autophagy is a conserved eukaryotic catabolic reaction that catabolizes cytoplasmic proteins and organelles. As a cytoprotective process, it helps maintain the homeostasis of cancer cells and acts as an alternative energy source in h...

Claims

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Application Information

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IPC IPC(8): A61K31/704A61P35/00A61K36/185
CPCA61K31/704A61K36/185A61P35/00
Inventor 张威王震应优敏李凯强张昱陈以栗郝珂
Owner ZHEJIANG PROVINCIAL PEOPLES HOSPITAL
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