New medical uses of Xanthium sibiricum

Abstract

The present invention relates to the field of medical technology, in particular to the field of natural medicines, and more specifically to a new medical use of Xanthate. The present invention first discovered that Xanthate can significantly inhibit the cell activity of BxPC-3 cells and PANC-1 cells, and this inhibitory effect shows a significant concentration dependence, and can significantly inhibit the cloning and division proliferation ability of BxPC-3 cells and PANC-1 cells, and Xanthate can also significantly inhibit the migration ability of BxPC-3 cells, and Xanthate can induce apoptosis of BxPC-3 cells, thereby providing a new idea for preparing a drug for treating pancreatic cancer. The drug preparation prepared by using Xanthate or Xanthate in combination with other drugs is safe and effective, and is a new pancreatic cancer treatment method.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, in particular to the field of natural medicines, and more specifically to a new medical application of Xanthium sibiricum. Background Art

[0002] Xanthatin is a sesquiterpenoid monomer component extracted from the traditional Chinese medicine Xanthium sibiricum. Its structural formula is as follows:

[0003]

[0004] Pancreatic cancer is a highly malignant digestive tract tumor. In recent years, the incidence of pancreatic cancer has been on the rise both at home and abroad. Due to the lack of early diagnostic methods, pancreatic cancer is mostly in the advanced metastatic stage when it is discovered, so its 5-year survival rate is only 9% to 10%. According to the American Cancer Society, the mortality rate of pancreatic cancer is expected to rank third among related malignant tumors in 2022, and its incidence rate is increasing year by year. According to data from the National Cancer Center of China, pancreatic cancer is expected to account for the sixth place in malignant tumor-related mortality in my country in 2022, and the incidence rate in men has increased significantly. In the past decade, the incidence and mortality rates of pancreatic cancer have increased significantly, and with the growth and aging of my country's population, the incidence of pancreatic cancer will become higher and higher in the future.

[0005] Pancreatic cancer is characterized by high malignancy, rapid development, and high mortality. According to the 2022 Pancreatic Cancer Diagnosis and Treatment Guidelines, the treatment of pancreatic cancer mainly includes surgery, radiotherapy, chemotherapy, intervention, immunotherapy, etc. Although surgery is the only way to cure pancreatic cancer, since most pancreatic cancers are in a locally advanced metastatic state when discovered, chemotherapy is still an important means of treating pancreatic cancer. In chemotherapy, according to the physical condition of pancreatic cancer patients, gemcitabine combined with albumin paclitaxel, cisplatin, capecitabine, and Segalovirus are generally used as first-line treatment options. However, chemotherapy drugs usually have toxic side effects on the human body. In addition, chemotherapy also faces the problem of drug resistance. Therefore, in order to improve the survival rate of pancreatic cancer patients and improve the quality of life of pancreatic cancer patients, it is particularly urgent to find safe and effective therapeutic drugs. Summary of the invention

[0006] The present invention aims to find a novel Chinese medicinal compound suitable for treating pancreatic cancer.

[0007] The present invention provides one of the new medical uses of Xanthium sibiricum, which refers to the use of Xanthium sibiricum in preparing a drug for inhibiting the activity of BxPC-3 cells.

[0008] At the same time, the present invention also provides a second new medical use of Xanthium sibiricum, which refers to the use of Xanthium sibiricum in the preparation of drugs for inhibiting the cloning and division ability of BxPC-3 cells.

[0009] The present invention also provides a third new medical use of Xanthium sibiricum, which refers to the use of Xanthium sibiricum in preparing a drug for inhibiting BxPC-3 cell migration.

[0010] The present invention also provides a fourth new medical use of Xanthium sibiricum, which refers to the use of Xanthium sibiricum in the preparation of a drug for inducing apoptosis of BxPC-3 cells.

[0011] In addition, the present invention also provides a fifth new medical use of Xanthium sibiricum, which refers to the use of Xanthium sibiricum in the preparation of a drug for inhibiting PANC-1 cell activity.

[0012] At the same time, the present invention also provides a sixth new medical use of Xanthium sibiricum, which refers to the use of Xanthium sibiricum in the preparation of drugs for inhibiting the cloning and division ability of PANC-1 cells.

[0013] Finally, the present invention also provides the use of Xanthium sibiricum in preparing medicine for treating pancreatic cancer.

[0014] A use of a pharmaceutical composition in preparing a drug for treating pancreatic cancer, wherein the pharmaceutical composition comprises Xanthium sibiricum.

[0015] The advantage of the present invention is that it is the first time to find that Xanthium sibiricum can significantly inhibit the cell activity of BxPC-3 cells and PANC-1 cells, and this inhibitory effect shows significant concentration dependence, and can significantly inhibit the cloning and division proliferation ability of BxPC-3 cells and PANC-1 cells, and Xanthium sibiricum can also significantly inhibit the migration ability of BxPC-3 cells, and Xanthium sibiricum can induce apoptosis of BxPC-3 cells, thereby providing a new idea for preparing drugs for treating pancreatic cancer. The drug preparation prepared by using Xanthium sibiricum or Xanthium sibiricum in combination with other drugs is safe and effective, and is a new pancreatic cancer treatment method. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Schematic diagram of the effects of different concentrations of Xanthium sibiricum on the viability of pancreatic cancer BxPC-3 cells.

[0017] Figure 2 Schematic diagram of the effects of different concentrations of Xanthium sibiricum on the viability of pancreatic cancer PANC-1 cells.

[0018] Figure 3 Schematic diagram of the effects of different concentrations of Xanthium sibiricum on the clone-forming ability of pancreatic cancer BxPC-3 cells.

[0019] Figure 4Schematic diagram of the effects of different concentrations of Xanthium sibiricum on the clone-forming ability of pancreatic cancer PANC-1 cells.

[0020] Figure 5 Schematic diagram of the effects of different concentrations of Xanthium sibiricum on the division ability of pancreatic cancer BxPC-3 cells.

[0021] Figure 6 Schematic diagram of the effects of different concentrations of Xanthium sibiricum on the cell division ability of pancreatic cancer PANC-1 cells.

[0022] Figure 7 This is a schematic diagram of the effects of different concentrations of Xanthium sibiricum on the cell migration ability of pancreatic cancer BxPC-3 cells after 0 to 24 hours.

[0023] Figure 8 Schematic diagram of the effects of different concentrations of Xanthium sibiricum on apoptosis of pancreatic cancer BxPC-3 cells. DETAILED DESCRIPTION

[0024] In order to better understand the present invention, the present invention is further described below in conjunction with specific embodiments.

[0025] In the following examples, Xanthium sibiricum was purchased from Huzhou Zhanshu Biotechnology Co., Ltd.; human pancreatic cancer cell lines BxPC-3 and PANC-1 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences; CCK was purchased from Dalian Meilun Biotechnology Co., Ltd.; crystal violet was purchased from Tianjin Damao Chemical Reagent Factory; CFDA SE cell proliferation and tracing detection kit was purchased from Shanghai Biyuntian Biotechnology Co., Ltd.; and AnnexinV-FITC apoptosis detection kit was purchased from BD Company of the United States.

[0026] Example 1

[0027] Human pancreatic cancer cell lines BxPC-3 cells and PANC-1 cells were inoculated into 96-well plates and cultured in a cell culture incubator. After the cells adhered to the wall and grew to a density of about 60% to 70%, the cells were treated with culture medium containing 0 μM, 10 μM, 20 μM, and 30 μM Xanthate at final concentrations for 24 hours, and the drug-containing culture medium was aspirated and discarded. The pre-prepared CCK-8 solution was added to each well of the 96-well plate at 100 μl, and incubated in an incubator for 1 to 2 hours. The OD value at 450 nm was measured with an enzyme marker and the cell viability was calculated. The experimental results refer to Figure 1 and Figure 2 .

[0028] The results showed that Xanthate could significantly inhibit the viability of pancreatic cancer BxPC-3 and PANC-1 cells, and with the increase of Xanthate concentration, its inhibitory effect on pancreatic cancer cell viability became stronger and dose-dependent. Compared with the blank group, the viability rates of BxPC-3 cells treated with 10μM, 20μM, and 30μM Xanthate were (74.92±2.56)%, (61.36±7.30)%, and (49.98±3.56)%, respectively, and the viability rates of PANC-1 cells treated with 10μM, 20μM, and 30μM Xanthate were (88.93±4.91)%, (47.06±4.97)%, and (40.07±3.32)%, respectively.

[0029] Example 2

[0030] Human pancreatic cancer cell lines BxPC-3 cells and PANC-1 cells were respectively plated at 3×10 3 The cells were inoculated at a density of 100 cells in a 6-well plate and placed in a cell culture incubator. After the cells adhered to the wall, the cells were treated with culture media containing final concentrations of 0 μM, 10 μM, 20 μM, and 30 μM Xanthate for 24 hours, and the drug-containing culture media was aspirated and discarded. Then, complete culture media containing 10% fetal bovine serum was added to the 6-well plate. The medium was changed every 2 to 3 days. When visible clones appeared in the 6-well plate, the culture was terminated, the culture media was aspirated and discarded, and the plate was washed twice with PBS and fixed with paraformaldehyde for 20 to 30 minutes. The paraformaldehyde was aspirated and discarded, and the plate was stained with crystal violet and photographed. The experimental results refer to Figure 3 and Figure 4 .

[0031] The results showed that Xanthium sibiricum could inhibit the cloning ability of pancreatic cancer BxPC-3 and PANC-1 cells in a concentration-dependent manner. Taking the number of clones in the blank group as the reference, that is, the number of clones in the blank group as 100%, compared with the blank group, the clone formation rates of BxPC-3 cells were 0.72±0.10, 0.29±0.09, and 0.060±0.02 when 10μM, 20μM, and 30μM Xanthium sibiricum were used, and the clone formation rates of PANC-1 cells were 0.85±0.03, 0.59±0.07, and 0.06±0.05 when 10μM, 20μM, and 30μM Xanthium sibiricum were used, respectively. It can be seen that under the action of Xanthium sibiricum, the clone formation rates of BxPC-3 cells and PANC-1 cells were significantly reduced.

[0032] Example 3

[0033] Human pancreatic cancer cell lines BxPC-3 and PANC-1 were respectively obtained, labeled with CFDA and inoculated into 6-well plates, and grown in a cell culture incubator to a density of about 60% to 70%. The cells were treated with culture medium containing Xanthate at final concentrations of 0 μM, 10 μM, 20 μM, and 30 μM, respectively, and then the culture was terminated after 24 hours. After the cells were collected, the cell fluorescence intensity was detected by flow cytometry. The experimental results refer to Figure 5 and Figure 6 .

[0034] The results of flow cytometry showed that with the increase of Xanthate concentration, the cell fluorescence value increased. Compared with the blank group, the increase ratio of 30 μM Xanthate to BxPC-3 cell CFDA fluorescence value was 1.37±0.10, and the increase ratio of 30 μM Xanthate to PANC-1 cell CFDA fluorescence value was 1.43±0.07, indicating that Xanthate can inhibit the cell division ability of pancreatic cancer cells.

[0035] Example 4

[0036] Human pancreatic cancer cell line BxPC-3 cells in logarithmic growth phase were inoculated into 6-well plates, placed in a cell culture incubator until the cells adhered to the wall and grew to a density of about 80% to 90%, and then horizontal scratches were made perpendicular to the 6-well plates with a 200 μl pipette tip. Culture medium containing Xanthium sibiricum at final concentrations of 0 μM, 10 μM, 20 μM, and 30 μM was added. Cell migration was observed by taking photos at 0 h, 12 h, and 24 h after drug addition. The experimental results are shown in Figure 7 .

[0037] Microscopic observations revealed that as the concentration and duration of Xanthate increased, the cell migration distance decreased, indicating that Xanthate can inhibit the migration ability of pancreatic cancer cells.

[0038] Example 5

[0039] Human pancreatic cancer cell line BxPC-3 cells were inoculated into 6-well plates and cultured in a cell culture incubator. After the cells adhered to the wall and grew to a density of about 60% to 70%, the culture was terminated after 24 hours of culture medium containing a final concentration of 0μM, 10μM, 20μM, and 30μM Xanthate. After trypsin digestion, the cells were collected by centrifugation, washed with pre-cooled PBS and resuspended, and 5μl of Annexin V-FITC staining solution and PI staining solution were added and mixed. The cells were placed at room temperature in the dark for staining, and the apoptosis of the cells was detected by flow cytometry after 15 minutes. The results are shown in Figure 8 .

[0040] The results of flow cytometer analysis showed that with the increase of Xanthate concentration, the Annexin V positivity of pancreatic cancer cells increased. The Annexin V positivity of cells under the action of 30 μM Xanthate was (22.72±4.60)%, which was significantly different from that of the blank group. The above results indicate that Xanthate can induce apoptosis of pancreatic cancer cells.

[0041] The above is a specific implementation of the present invention. It should be noted that, for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications are also considered to be within the protection scope of the present invention.

Claims

1. Application of Xanthium sibiricum in the preparation of drugs for treating pancreatic cancer.

2. Use of a pharmaceutical composition in the preparation of a drug for treating pancreatic cancer, characterized in that: The pharmaceutical composition includes Xanthium sibiricum.

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