CN111374979A

CN111374979A - Medical application of compound in treatment of oral cancer by inducing apoptosis of oral cancer cells

Info

Publication number: CN111374979A
Application number: CN202010198394.1A
Authority: CN
Inventors: 孙强; 郑宏雨; 李紫璇; 陈怡恒; 赵军方; 薛文华; 李光辉; 孙明磊
Original assignee: First Affiliated Hospital of Zhengzhou University
Current assignee: First Affiliated Hospital of Zhengzhou University
Priority date: 2020-03-19
Filing date: 2020-03-19
Publication date: 2020-07-07
Anticipated expiration: 2040-03-19
Also published as: CN111374979B

Abstract

The invention discloses a medical application of a compound in treating oral cancer by inducing apoptosis of oral cancer cells. The invention discovers that the two chalcone derivatives can inhibit the growth of oral cancer by inhibiting the proliferation and inducing the apoptosis of oral cancer cells on one hand and can inhibit the migration activity of the oral cancer cells on the other hand. Therefore, the two chalcone derivatives have the prospect of being developed into medicines for resisting the growth of oral cancer and resisting the metastasis of the oral cancer, and are used for treating the oral cancer.

Description

Medical application of compound in treatment of oral cancer by inducing apoptosis of oral cancer cells

Technical Field

The invention belongs to the field of medicines, relates to screening discovery of oral cancer medicines, and particularly relates to a medical application of a compound in treating oral cancer by inducing apoptosis of oral cancer cells.

Background

Oral cancer is a common head and neck tumor with a high incidence rate. In recent years, with the rapid development of global economy and the improvement of the living standard of people, the incidence rate of oral cancer is remarkably increased, and the incidence population tends to be younger.

At present, surgical treatment is the most effective treatment method in clinic, but oral cancer is relatively hidden and is often found in the late stage, so that the best time for surgical treatment is missed. In addition, chemotherapy is also an important treatment method for oral cancer, and therapeutic drugs include fluorouracil, cisplatin and the like, but the existing chemotherapy method not only has great toxic and side effects, but also is easy to generate drug resistance. Therefore, the search for high-efficiency and low-toxicity oral cancer prevention and treatment medicines is still a serious task for researchers.

Chalcone and derivatives thereof are α -unsaturated carbonyl-containing compounds, the natural chalcone compounds mainly exist in natural plants such as liquorice and the like, and have small toxic and side effects and high reaction activity, so that the chalcone and the derivatives thereof become important chemical raw materials and medical intermediates and are used for screening and discovering medicaments.

The application of chalcone derivatives with the following chemical structures in preparing anti-oral cancer drugs is not reported at present.

Disclosure of Invention

The invention aims to provide a medical application of a compound in treating oral cancer by inducing apoptosis of oral cancer cells.

The above purpose of the invention is realized by the following technical scheme:

the medical application of chalcone derivative with the following chemical structure in preparing medicines for treating oral cancer is as follows:

the chalcone derivative is used for preparing medicines for promoting the apoptosis of oral cancer and inhibiting the growth of the oral cancer.

The chalcone derivative is used for preparing medicines for inhibiting oral cancer migration.

Furthermore, the medicine takes the chalcone derivative as a medicinal component and also contains pharmaceutically acceptable auxiliary materials, and is prepared into pharmaceutically acceptable dosage forms.

Further, the adjuvant is a liquid, solid or semi-solid adjuvant.

Still further, the dosage forms include tablets, capsules, injections.

Has the advantages that:

the invention discovers that two chalcone derivatives with the following chemical structures can inhibit the growth of oral cancer by inhibiting the proliferation and inducing the apoptosis of oral cancer cells on one hand and can inhibit the migration activity of the oral cancer cells on the other hand. Therefore, the two chalcone derivatives have the prospect of being developed into medicines for resisting the growth of oral cancer and resisting the metastasis of the oral cancer, and are used for treating the oral cancer.

Drawings

FIG. 1 shows the chemical structures of chalcone derivative 1 and chalcone derivative 2;

fig. 2 is an inhibition curve of chalcone derivative 1 on human oral cancer KB cells, with drug concentration on the abscissa and inhibition rate on the ordinate; the results show that the chalcone derivative 1 has a remarkable inhibition effect on the proliferation activity of human oral cancer KB cells, and the proliferation inhibition effect presents a relatively obvious dose-effect relationship;

fig. 3 is an inhibition curve of chalcone derivative 2 on human oral cancer KB cells, with drug concentration on the abscissa and inhibition rate on the ordinate; the results show that the chalcone derivative 2 has a remarkable inhibition effect on the proliferation activity of human oral cancer KB cells, and the proliferation inhibition effect presents a relatively obvious dose-effect relationship;

FIG. 4 shows the apoptosis rates of the various groups; from the results, the apoptosis rate of the chalcone derivatives 1 and 2 is significantly higher than that of the negative control group, which indicates that the chalcone derivatives 1 and 2 have the activity of inducing apoptosis of human oral cancer KB cells;

FIG. 5 shows the number of migrating cells in each group; from the results, it can be seen that the cell migration number of the chalcone derivatives 1 and 2 groups was significantly lower than that of the negative control group, indicating that the chalcone derivatives 1 and 2 have the activity of inhibiting the migration of human oral cancer KB cells.

Detailed Description

The following detailed description of the present invention is provided in connection with the accompanying drawings and examples, but not intended to limit the scope of the invention.

First, experimental material

The human oral cancer KB cells are frozen in liquid nitrogen and recovered by a conventional method for use.

Fetal bovine serum, RPMI1640 medium, trypsin under the brand Gibco.

The brands of penicillin and streptomycin are Sigma, and the PBS solution is prepared according to the formula. The purity of the chalcone derivative 1 and the chalcone derivative 2 is more than or equal to 98 percent, and the chemical structures are shown in figure 1.

Cell culture plates were purchased from Thermo Scientific.

Transwell chambers (24 wells) were purchased from Corning.

The CCK-8 kit was purchased from Homophilus Japan.

The brand of the annexin V-FITC/PI apoptosis detection kit is Solarbio.

Second, Experimental methods

1. Cell recovery and subculturing

Taking out human oral cancer KB cells from a liquid nitrogen tank, placing the human oral cancer KB cells in a water bath kettle at 37 ℃, slightly shaking the human oral cancer KB cells for melting, blowing and uniformly mixing the human oral cancer KB cells by using a pipette gun after melting, transferring the human oral cancer KB cells into a 15mL centrifuge tube, adding a proper amount of complete culture solution (containing 10% of embryo)RPMI1640 medium of bovine serum, 100U/mL penicillin, 100. mu.g/mL streptomycin), centrifuging, discarding supernatant, adding appropriate amount of complete culture solution to resuspend cells, transferring to cell culture flask, placing at 37 deg.C and 5% CO₂Culturing in incubator, removing culture medium when cell bottle is full of cells, washing with PBS, digesting cells with 0.25% trypsin, centrifuging when cell gap is enlarged, removing supernatant, washing with PBS, adding complete culture medium, inoculating into new cell bottle, placing at 37 deg.C and 5% CO₂Culturing in an incubator and carrying out passage. Well-grown cells in logarithmic growth phase were collected for experiments.

2. Cell grouping and drug delivery culture

The experimental groups included:

(1) negative control group: culturing with complete culture medium without medicine;

(2) chalcone derivatives group 1 (derivatives group 1): culturing with complete medium containing 20, 50, 100, 200, 500nM chalcone derivative 1;

(3) chalcone derivatives group 2 (derivatives group 2): the cells were cultured in complete medium containing 20, 50, 100, 200, 500nM chalcone derivative 2, respectively.

3. CCK-8 method for measuring cell proliferation activity

Digesting human oral cancer cells in logarithmic growth phase with 0.25% trypsin to obtain cell suspension, and adjusting cell density to 5 × 10⁴and/mL, transferring the cell suspension to a 96-well plate, wherein each well is 200 mu L, and pre-culturing for 24h until the cells adhere well. Grouping according to the '2, cell grouping and administration culture', each group comprises 6 multiple wells, after culturing for 48h, absorbing and discarding the original culture solution, adding 100 mu L of fresh culture medium and 10 mu L of CCK-8 reaction solution into each well, incubating for 4h, measuring the absorbance value (OD) of each group at 450nm by using an enzyme-labeling instrument, calculating the inhibition rate of the drug on human oral cancer cells according to the following formula, and drawing a concentration-inhibition rate curve.

Inhibition rate (1-OD drug/OD negative control) × 100%

4. Determination of apoptosis rate by flow cytometry

Taking human oral cancer cells in logarithmic growth phase, digesting with 0.25% trypsinForming cell suspension, adjusting cell density to 5 × 10⁴and/mL, transferring the cell suspension to a 96-well plate, wherein each well is 200 mu L, and pre-culturing for 24h until the cells adhere well. Grouping according to the '2, cell grouping and administration culture' (only 50nM and 100nM concentrations are set for the drug group), repeating the hole for 6 each group, after culturing for 48h, respectively adding PI and Annexin V/FITC in 5 muL under the condition of keeping out of the sun, after 20min, strictly according to the procedures of an Annexin V-FITC/PI apoptosis detection kit operation instruction, and measuring the apoptosis rate of the human oral cancer KB cell by using a flow cytometer.

5. Transwell method for measuring cell migration activity

Collecting human oral cancer cells in logarithmic growth phase, digesting, centrifuging, culturing with complete culture medium containing or not containing drug for 48h (negative control group contains no drug, and derivative 1 group and derivative 2 group respectively contain 20nM drug), starving with serum-free culture medium for 12h, digesting, centrifuging, re-suspending with serum-free culture medium, counting, adding 200 μ L cell suspension into each upper chamber, each well containing 2 × 10⁵And (4) cells. Putting the whole small chamber into a 24-pore plate containing complete culture medium for culturing for 24h, taking out the upper chamber, fixing with anhydrous methanol, staining with crystal violet, wiping off the upper surface without membrane cells, sealing, observing with an inverted microscope, randomly selecting 5 visual fields, counting under a high power microscope, and averaging.

6. Statistical treatment

Processing was performed using SPSS version 17.0 software. Data are presented as mean ± standard deviation, with t-test for inter-group comparisons and one-way anova for inter-group comparisons. The difference is statistically significant when P is less than 0.05.

Third, experimental results

1. Effect of chalcone derivatives on cell proliferative Activity

The results are shown in table 1 and fig. 2-3, the chalcone derivatives 1 and 2 can obviously inhibit the proliferation activity of human oral cancer KB cells, and the proliferation inhibition effect presents a relatively obvious dose-effect relationship.

TABLE 1 inhibition of proliferation of human oral cancer KB cells by chalcone derivatives 1 and 2

2. Effect of chalcone derivatives on apoptosis

The results are shown in table 2 and fig. 4, and it can be seen from the results that the apoptosis rate of chalcone derivatives 1 and 2 group is significantly higher than that of the negative control group, indicating that chalcone derivatives 1 and 2 have the activity of inducing apoptosis of human oral cancer KB cells.

TABLE 2 apoptosis rates of the groups

3. Effect of drugs on cell migration Activity

As shown in table 3 and fig. 5, it can be seen from the results that the cell migration number of chalcone derivatives 1 and 2 was significantly lower than that of the negative control group, indicating that chalcone derivatives 1 and 2 have the activity of inhibiting the migration of human oral cancer KB cells.

TABLE 3 number of migrating cells in each group

In conclusion, chalcone derivatives 1 and 2 can inhibit the growth of oral cancer by inhibiting the proliferation and inducing the apoptosis of oral cancer cells, and can inhibit the migration activity of oral cancer cells. Therefore, the two chalcone derivatives have the prospect of being developed into medicines for resisting the growth of oral cancer and resisting the metastasis of the oral cancer, and are used for treating the oral cancer.

The above-described embodiments are intended to be illustrative of the nature of the invention, but those skilled in the art will recognize that the scope of the invention is not limited to the specific embodiments.

Claims

1. The medical application of chalcone derivative with the following chemical structure in preparing medicines for treating oral cancer is as follows:

2. the use of the chalcone derivative according to claim 1 for preparing a medicament for promoting apoptosis and inhibiting growth of oral cancer.

3. The use of a chalcone derivative according to claim 1 for the preparation of a medicament for inhibiting metastasis of oral cancer.

4. The pharmaceutical use according to any one of claims 1 to 3, wherein: the medicine takes the chalcone derivative as a medicine effect component and also contains pharmaceutically acceptable auxiliary materials, and is prepared into pharmaceutically acceptable dosage forms.

5. The medical use according to claim 4, wherein: the auxiliary material is liquid, solid or semisolid auxiliary material.

6. The medical use according to claim 4, wherein: the dosage forms comprise tablets, capsules and injections.