Application of lupenone in preparation of medicine for treating colorectal cancer

Abstract

The invention belongs to the field of colorectal cancer treatment, and particularly relates to application of lupenone in preparation of a medicine for treating colorectal cancer. The research finds that the lupenone has an excellent treatment effect on the colorectal cancer: on one hand, the lupenone can directly kill a colorectal cancer cell line CT26, inhibit the growth of in-vivo and in-vitro CT26 cells and reduce the growth of tumors; on the other hand, the lupenone is combined with the chemotherapeutic drug for use, so that the anti-tumor effect of the chemotherapeutic drug can be enhanced. Therefore, lupenone can be used as a traditional Chinese medicine compound to be applied to clinical treatment of colorectal cancer, and has a good application prospect.

Description

Technical Field

[0001] The invention belongs to the field of colorectal cancer treatment, and specifically relates to an application of lupine ketone in the preparation of a drug for treating colorectal cancer. Background Art

[0002] Colorectal cancer is one of the most common malignant tumors worldwide, and its incidence and mortality rates are increasing year by year. The incidence of colorectal cancer is more common in people over 50 years old, and the proportion of young patients is gradually increasing, which may be related to lifestyle, eating habits and genetic factors. Because the early symptoms are not obvious, many patients are already in the late stage when diagnosed, which greatly increases the difficulty. At present, the main treatments for colorectal cancer are surgery and chemotherapy, but the treatment effect is poor, which makes the treatment of colorectal cancer face huge challenges. Therefore, while improving surgical treatment, finding new anti-tumor drugs that are safe, efficient and non-resistant has become the focus of attention.

[0003] my country has a vast territory and rich resources of Chinese medicinal materials. If compounds that can be used to fight colorectal cancer can be extracted from Chinese medicine, it will be of great significance for the treatment of colorectal cancer. Musaba root (scientific name: MusabasjooSieb.etZucc.) is a Chinese medicine name. It belongs to the Musabas order, Musaceae, and Musa genus. It is distributed in Fujian, Guangdong, Hainan, Guangxi, Yunnan and other places. The rhizome is used as medicine, with a light nature and cool taste. It has the effects of clearing heat, quenching thirst, diuresis, and detoxification. It is often used for natural fever, depression, thirst, jaundice, edema, beriberi, hematuria, hemorrhage, carbuncle, furuncle, and erysipelas.

[0004] Lupeneone (Lup) is a compound extracted from banana roots and has great pharmacological potential. Currently, there is no report on the use of lupeneone in the treatment of colorectal cancer. Based on this, the technical solution of the present invention is proposed. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the present invention provides an application of lupine ketone in preparing a drug for treating colorectal cancer.

[0006] Preferably, the colorectal cancer cells are CT26 cells.

[0007] Preferably, the drug is a drug that reduces the viability of CT26 cells.

[0008] Preferably, the drug is a drug that inhibits tumor cell growth.

[0009] Preferably, the drug is a drug that causes an increase in intracellular ROS levels.

[0010] Preferably, the drug is a drug that causes an increase in the level of ERS in cells.

[0011] Preferably, the drug is a drug that causes increased intracellular autophagy and autophagic flux disorder.

[0012] Preferably, the drug is a preparation prepared by using lupine ketone as a raw material and adding pharmaceutically acceptable excipients.

[0013] Preferably, the preparation is an oral preparation or an injectable preparation.

[0014] Based on the same technical concept, the present invention further provides an application of lupine ketone and cisplatin in the preparation of drugs for treating colorectal cancer.

[0015] The beneficial effects of the present invention are: The present invention has found that lupine ketone has an excellent therapeutic effect on colorectal cancer: on the one hand, lupine ketone can directly kill the colorectal cancer cell line CT26, inhibit the growth of CT26 cells in vivo and in vitro, and reduce the growth of tumors; on the other hand, lupine ketone combined with chemotherapy drugs can enhance the anti-tumor effect of chemotherapy drugs. Therefore, lupine ketone can be used as a traditional Chinese medicine compound in the treatment of clinical colorectal cancer, and has a good application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0017] Figure 1 This is the effect of Lupenone on the viability of CT26 cells; where: *p<0.05, **p<0.01 indicate statistical significance compared with the Lup-0 μM group.

[0018] Figure 2 This is a graph showing the effect of Lupenone on the growth of CT26 cell clones.

[0019] Figure 3 This is a graph showing the effect of Lupenone on ROS in CT26 cells.

[0020] Figure 4 The figure is the effect of Lupenone on the expression of ERS-related genes Grp78 and Chop in CT26 cells; where: *p<0.05, **p<0.01 indicate statistical significance compared with the Lup-0 μM group.

[0021] Figure 5The figure shows the effect of Lupenone on the expression of autophagy-related proteins p62 and LC3B (including LC3I and LC3II) in CT26 cells; among them: *p<0.05, **p<0.01 indicate statistical significance compared with the Lup-0 μM group.

[0022] Figure 6 This is a graph showing the effect of Lupenone on colon cancer tumor growth.

[0023] Figure 7 Graph showing the effect of lupetone and cisplatin combination therapy on colon cancer tumor growth.

[0024] Figure 8 Volcano plots of differential gene expression and gene enrichment analysis results of tumor tissue transcriptome sequencing in the cisplatin group and the cisplatin + lupine ketone combined treatment group; among which: Figure 8 A in the figure is a volcano map; Figure 8 B in the figure is the enrichment diagram of IL17 signaling pathway; Figure 8 C in the figure is the enrichment diagram of inflammatory bowel disease pathway; Figure 8 D in the figure is the enrichment diagram of TNF signaling pathway; Figure 8 E in the figure is a diagram of TH1 and TH2 cell differentiation and enrichment; Figure 8 F in the figure is a diagram showing the differentiation and enrichment of TH17 cells. DETAILED DESCRIPTION

[0025] To make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation methods obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0026] Example 1

[0027] Colorectal cancer cell line CT26 cells were cultured, 10,000 CT26 cells were seeded in a 96-well plate, different concentrations of lupeone (Lup) were added (0, 10, 20, 40 μM), and treated for 24 hours. Cell viability was detected using the cell proliferation detection kit CCK8. Figure 1 As shown, the results showed that lupeolone could reduce the viability of CT26 cells in a concentration-dependent manner.

[0028] Example 2

[0029] Culture colorectal cancer cell line CT26 cells, inoculate 500 CT26 cells in a 6-well plate, add different concentrations of lupeone (Lup) (0, 20, 40 μM), change the culture medium every 3 days, culture for 10 days, fix the cells with paraformaldehyde, perform crystal violet staining, and take pictures to observe the cell mass. Figure 2 As shown, the results showed that lupeolone could significantly kill CT26 cells and inhibit the growth of tumor cells.

[0030] Example 3

[0031] CT26 colorectal cancer cell line was cultured and 10×10 4 Different concentrations of lupeone (Lup) were added to CT26 cells (0, 20, 40 μM) and cultured for 24 hours. The intracellular ROS level was detected using a ROS detection kit. Figure 3 As shown, the results showed that lupeone caused an increase in intracellular ROS levels.

[0032] Example 4

[0033] CT26 colorectal cancer cell line was cultured and 10×10 4 CT26 cells were treated with different concentrations of lupeone (Lup) (0, 20, 40 μM) and cultured for 24 h. The cells were collected and RNA was extracted to detect the expression of endoplasmic reticulum stress (ERS) related genes Grp78 and Chop. Figure 4 As shown, the results showed that lupeone caused an increase in the intracellular ERS level.

[0034] Example 5

[0035] CT26 colorectal cancer cell line was cultured and 10×10 4 CT26 cells were treated with different concentrations of lupeinone (Lup) (0, 20, 40 μM) and cultured for 24 hours. Cell proteins were collected and the expressions of autophagy-related proteins LC3B and p62 were detected. Figure 5 As shown, the results indicated that lupetone caused increased intracellular autophagy and impairment of autophagic flux.

[0036] Example 6

[0037] Colorectal cancer cell line CT26 cells were cultured and injected subcutaneously into the right axilla of mice with 20×10 4 The mice were randomly divided into two groups: a control group (treated with saline) and a lupine ketone treatment group (16 mg / Kg). The drug was injected for 14 days, and the growth of tumors in the two groups was observed. Figure 6 As shown, the results showed that lupeolone could significantly inhibit the growth level of tumors.

[0038] Example 7

[0039] Colorectal cancer cell line CT26 cells were cultured and injected subcutaneously into the right axilla of mice with 20×10 4 CT26 cells were randomly divided into three groups: control group, cisplatin group (3 mg / Kg), and cisplatin (3 mg / Kg) + lupine ketone (16 mg / Kg) combined treatment group. The drugs were injected for 14 days, and the growth of tumors in the three groups was observed. Figure 7 As shown, the results indicate that lupeone can enhance the antitumor effect of cisplatin.

[0040] Example 8

[0041] Tumor tissues from Experiment 7 were taken out for transcriptomic sequencing to analyze differentially expressed genes and gene enrichment. Figure 8 As shown, the results indicate that lupine ketone regulates immune cell differentiation and inflammatory signaling pathways, affects the tumor immune microenvironment, and enhances the anti-tumor effect of cisplatin.

[0042] in conclusion: (1) Lupeone (Lup) can effectively inhibit the growth of colorectal cancer tumors, mainly by regulating the cellular autophagy pathway triggered by the ROS-ERS cascade in CT26 cells.

[0043] (2) Lupeneone (Lup) combined with chemotherapy drugs can enhance the anti-tumor effect of chemotherapy drugs, mainly by regulating the differentiation of Th1, Th2, and Th17 cells and regulating the tumor immune microenvironment.

[0044] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. Application of lupine ketone in the preparation of drugs for the treatment of colorectal cancer.

2. The use according to claim 1, characterized in that: The colorectal cancer cells are CT26 cells.

3. The use according to claim 2, characterized in that: The drug is a drug that reduces the activity of CT26 cells.

4. The use according to claim 2, characterized in that: The drug is a drug that inhibits the growth of tumor cells.

5. The use according to claim 2, characterized in that: The drug is a drug that causes an increase in the level of ROS in cells.

6. The use according to claim 2, characterized in that: The drug is a drug that causes an increase in the level of ERS in cells.

7. The use according to claim 2, characterized in that: The drug is a drug that causes increased autophagy in cells and autophagy flow disorder.

8. The use according to any one of claims 1 to 7, characterized in that: The medicine is a preparation prepared by taking lupine ketone as a raw material and adding pharmaceutically acceptable auxiliary materials.

9. The use according to claim 8, characterized in that: The preparation is an oral preparation or an injection preparation.

10. Application of lupine ketone and cisplatin in the preparation of drugs for the treatment of colorectal cancer.

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