Application of subprostrate as inducer for inhibiting autophagy of thyroid undifferentiated cancer cells

Abstract

The invention discloses an application of subprostrate sophora as an autophagy inducer for undifferentiated thyroid cancer cells, and the subprostrate sophora can promote autophagy of the undifferentiated thyroid cancer cells by inhibiting AKT / mTOR pathway, inhibit proliferation of the undifferentiated thyroid cancer cells and further promote death of tumor cells. A new theoretical and experimental basis is provided for treating the thyroid undifferentiated cancer, a brand-new anti-tumor way for the thyroid undifferentiated cancer is provided at the same time, and the potential of the subprostrate as a clinical auxiliary drug is provided.

Description

(I) Technical field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of shantonin as an autophagy inducer for anaplastic thyroid cancer cells. (II) Background technology

[0002] Thyroid cancer is a common malignant tumor of the endocrine system, and its incidence has increased significantly in recent years. Among thyroid cancers, anaplastic thyroid carcinoma (ATC) has the worst prognosis and is the most aggressive subtype of thyroid cancer. The median survival of ATC is only 4-6 months. Although its incidence accounts for only about 5% of the total number of thyroid tumors, it is extremely deadly and prone to recurrence. There is currently no effective treatment for patients with distant metastasis. In clinical practice, surgical resection, targeted therapy and radiotherapy can benefit patients with relatively good health, but the mortality rate of cancer recurrence is still close to 100%.

[0003] my country has rich resources of traditional Chinese medicine. As a new source of anti-cancer drugs that are gaining more and more attention, Chinese medicine has the advantages of few toxic side effects, multiple therapeutic targets, and improving the adverse reactions of tumor chemotherapy. With the discovery of active ingredients of Chinese medicine and the continuous exploration of anti-tumor mechanisms, the integration of Chinese and Western medicine is becoming a new trend in cancer prevention and treatment in modern society.

[0004] Sophoranone, also known as Guangdougen, is the dried root and rhizome of the leguminous plant Sophora tonkinensis Gagnep. It is mainly produced in Yunnan, Guizhou, Guangxi and other places. It has the effects of clearing away heat and detoxifying, reducing swelling and relieving sore throat. Sophoranone is a natural compound belonging to the isoflavone class of compounds. It is extracted from Sophora tonkinensis and is widely used in traditional Chinese medicine formulas. It has anti-inflammatory, antibacterial, antioxidant, anti-tumor and other pharmacological effects. There are literature reports that Sophoranone has found significant anti-tumor ability in liver cancer cells, but there have been few reports on Sophoranone in recent years, and there has never been any report on its effect in anaplastic thyroid cancer.

[0005] Autophagy is an important process of intracellular material degradation and metabolism. Misfolded, damaged, denatured or aged proteins or organelles are wrapped in a double membrane and then fused with lysosomes. Under the action of lysosomal acid hydrolases, they are broken down into small molecules such as amino acids and recycled and reused by cells. In the early stage of tumorigenesis, autophagy can maintain cell homeostasis to inhibit the occurrence and development of tumors. In addition, autophagy can promote tumor apoptosis and inhibit the migration and invasion of tumor cells. In many mouse models, knocking out autophagy-related genes leads to accelerated tumor development in mice. Therefore, promoting autophagy may be an auxiliary means of treating tumors. It has been reported that autophagy agonists such as rapamycin can inhibit the expression of the PI3K / AKT / mTOR pathway, promote autophagy, help tumor cells form autophagosomes, and enhance tumor cell apoptosis. Therefore, further exploring the specific mechanism of drug-induced autophagy in anaplastic thyroid cancer will help provide more accurate prevention and treatment strategies for cancer patients. (III) Summary of the invention

[0006] The purpose of the present invention is to provide a use of scutellaria baicalensis as an autophagy inducer for anaplastic thyroid cancer cells. Scutellaria baicalensis can promote the autophagy of anaplastic thyroid cancer cells by inhibiting the AKT / mTOR pathway, inhibit the proliferation of anaplastic thyroid cancer cells, and then promote the death of tumor cells.

[0007] The technical solution adopted by the present invention is:

[0008] The present invention provides an application of scutellaria baicalensis as an autophagy inducer for anaplastic thyroid cancer. The inducer can promote the autophagy of anaplastic thyroid cancer cells.

[0009] Furthermore, the inducer promotes autophagy of anaplastic thyroid cancer cells by inhibiting the AKT / mTOR pathway, thereby promoting the death of tumor cells.

[0010] The present invention also provides an application of scutellaria baicalensis in preparing a medicine for treating anaplastic thyroid cancer.

[0011] Furthermore, the drug can inhibit the proliferation of anaplastic thyroid cancer cells.

[0012] The structural formula of sophora indica is:

[0013]

[0014] Compared with the prior art, the beneficial effects of the present invention are mainly reflected in:

[0015] The present invention provides a new application of shantonin as an autophagy inducer for anaplastic thyroid cancer. Shantonin can regulate tumor cell autophagy by inhibiting the AKT / mTOR pathway and inhibit the proliferation of anaplastic thyroid cancer cells, providing a new theoretical and experimental basis for the treatment of anaplastic thyroid cancer. It also provides a new anti-tumor pathway for anaplastic thyroid cancer and the potential of shantonin as a clinical auxiliary drug. (IV) Description of the drawings

[0016] Figure 1 The inhibitory effect of shantonin on thyroid cancer cell lines; A represents the relative cell viability curve of different thyroid cancer cell lines after treatment with shantonin; B represents the microscopic images of the scratch test of 8505C and KMH-2 cells after treatment with shantonin; C represents the microscopic images of the cloning test of different thyroid cancer cell lines after treatment with shantonin; D represents the microscopic images of the migration test of 8505C and KMH-2 cells after treatment with shantonin.

[0017] Figure 2 The experimental results of shantonin-induced autophagy in ATC cells; Figure A represents the cell viability of 8505C cells and KMH-2 cells after pretreatment with necroptosis inhibitor Necrostatin-1 (Nec-1) and apoptosis inhibitor Z-VAD-FMK (Z-Vad-Fmk) and then reversing shantonin; Figure B represents the micrograph of 8505C cells and KMH-2 cells after treatment with shantonin; Figure C represents the cell viability of 8505C cells and KMH-2 cells after pretreatment with autophagy inhibitor chloroquine (CQ) and then reversing shantonin; Figure D represents the fluorescence staining image after treatment with shantonin.

[0018] Figure 3 These are the experimental results of studying the effects of shantonin on autophagy-related proteins in ATC cells. (V) Specific implementation methods

[0019] The present invention is further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited thereto:

[0020] The methods are conventional methods unless otherwise specified. The raw materials can be obtained from public commercial channels unless otherwise specified. The quantitative tests in the following examples are repeated three times and the results are averaged. The room temperature in the present invention is 25-30°C.

[0021] Example 1. Materials and methods

[0022] 1. Drugs and reagents

[0023] Sophora flavescens was purchased from Sichuan Cuiyirun Biotechnology Co., Ltd., and dimethyl sulfoxide (DMSO) was purchased from BBI (Shanghai, China). Sophora flavescens was prepared with DMSO in the form of a 100 mM stock solution.

[0024] CCK-8 kit was purchased from Novagen Biotech. ER-Tracker Red (endoplasmic reticulum red fluorescent probe), Lyso-Tracker Red (lysosome red fluorescent probe), Mito-Tracker Green (mitochondrial green fluorescent probe), Western and IP cell lysate, BCA protein concentration assay kit, 5X Evo M-MLV RT Master Mix were all purchased from Shanghai Bio-Tech Biotechnology Co., Ltd.; primary antibody Phospho-mTOR (Ser2448) (D9C2) Rabbit mAb #5536 (referred to as p-mTOR), Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060 (referred to as p-AKT473), Phospho-Akt (Thr308) (D25E6) Rabbit mAb#13038 (referred to as p-AKT308), Phospho-4E-BP1(Thr37 / 46)(236B4)Rabbit mAb#2855 (referred to as p-4EBP1), β-Actin(D6A8)Rabbit mAb#8457, Phospho-S6Ribosomal Protein(Ser235 / 236)

[0025] Antibody #2211 (referred to as p-S6K S235 / 236), LC3B Antibody #2775 (including LC3B-Ⅰ / LC3B-Ⅱ) and goat anti-rabbit and goat anti-mouse secondary antibodies (lot number 7074S) were purchased from CST, USA. The rest of the consumables were purchased from Bio Sharp.

[0026] Cell lines

[0027] Human anaplastic thyroid cancer cell 8505C was obtained from the German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen), and human anaplastic thyroid cancer cell lines BHT101, CAL62, and KMH-2 were purchased from Wuhan Procell Life Science & Technology Co., Ltd. All of the above cells were STR certified and cultured in HyClone RPMI-1640 medium (hereinafter referred to as complete medium) containing 10% FBS, 100U / mL penicillin, and 0.1mg / mL streptomycin, and were placed in an incubator with a CO2 concentration of 5% and a temperature of 37°C.

[0028] 2. Methods

[0029] 2.1 Effect of shantonin on the activity of thyroid cancer cells

[0030] (1) CCK-8 assay to detect cell viability

[0031] Human anaplastic thyroid cancer cells 8505C, BHT101, CAL62, and KMH-2 in logarithmic growth phase were selected and the cell count of each cell was 2×10 3 The number of cells was inoculated in a 96-well cell culture plate containing 100 μL of complete medium in each well. After the cells adhered, the supernatant was discarded, and 200 μL of complete medium containing 0, 20, 25, 30, 35, 40, 45, 50, 55, and 60 μM shantonin was added to the corresponding wells of the 96-well plate. In addition, 200 μL of complete medium containing DMSO with an equal volume of 60 μM shantonin was set up as a control group, and 200 μL of complete medium was added as a blank control group, with 3 replicates in each group. After the 96-well plate was placed in an incubator with a concentration of 5% CO2 and a temperature of 37°C for 48 hours, 10 μL of CCK-8 reagent was added to each well and placed in an incubator with a concentration of 5% CO2 and a temperature of 37°C for incubation for 2 hours; the absorbance value (A) of each well was measured using an enzyme reader at a wavelength of 450 nm. The survival rate of all cells at 48 hours was calculated, and the survival rate % = (A 药物组 -A 空白组 ) / (A 对照组 -A 空白组 )×100%, the experiment was repeated 3 times.

[0032] (2) Scratch test

[0033] First, use a pen and ruler to draw parallel lines evenly on the back of the 6-well plate. The spacing between each horizontal line is 0.5-1 cm. Draw 3 horizontal lines in each well to facilitate the observation of each area under the microscope. Take anaplastic thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase and count the cells at 2×10 5 The amount of cells / well was inoculated in a 6-well plate supplemented with 2000μL complete medium. After the cells adhered to the wall, a micropipette tip was used to draw a line in the central area of ​​cell growth so that the scratch and the bottom line intersected, and the cells in the central part were removed. Each well of the 6-well plate was divided into a control group, a 30μM drug group, and a 35μM drug group. 2000μL of complete medium was added to the control group, and 2000μL of complete medium containing 30μM and 35μM shantonin was added to the drug group, respectively, with 3 replicates in each group. Place in an incubator with a 5% CO2 concentration and a temperature of 37°C and culture for 0, 12, and 24h. The gaps between the cells were photographed at 0, 12, and 24h, respectively, and the cells were imaged using the cell imaging system EVOS TM Imaging and shooting in the M7000 imaging system.

[0034] (3) Clone formation experiment

[0035] Undifferentiated thyroid cancer cells 8505C, BHT101, CAL62 and KMH-2 in the logarithmic growth phase were collected and the cells were counted and the number of cells was 5×10 3 The amount of cells / well was inoculated in a 6-well plate with 5000μL complete medium, and the culture medium was discarded after the cells adhered to the wall. Set up a control group and a drug group. The drug group added 5000μL of complete medium containing 25μM, 30μM, and 35μM shantonin, and the control group added 5000μL of complete medium, with 3 replicates in each group. Each group was placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for about 14 days. When obvious colonies were formed in the 6-well plate, the culture was stopped and the old culture medium was discarded. After washing with PBS twice, 4% tissue fixative was added to fix the cells at room temperature for 20 minutes. After discarding the fixative, wash with PBS twice, add crystal violet to stain the cells for 20 minutes, and wash with PBS twice and discard the liquid. After air drying at room temperature, use Deli 15161S high-speed camera A3 soft pad (gray) (table) to photograph the colonies.

[0036] (4) Migration and invasion assay

[0037] 30 minutes before the invasion experiment, 8 permeable cell culture chambers were placed in a 24-well plate, of which the upper chambers of 4 chambers were added with Mogengel Matrix and HyClone RPMI-1640 medium at a volume ratio of 1:4. The 4 chambers with matrix gel were used for invasion experiments (1 for the control group and 3 for the experimental group), and the remaining 4 chambers were used for migration experiments (1 for the control group and 3 for the experimental group). Cell migration and invasion experiments were performed in groups of 8 chambers.

[0038] Undifferentiated thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase were obtained. After digestion, the cells were uniformly resuspended in HyClone RPMI-1640 medium and counted. 5×10 4 The number of cells was inoculated in the upper chamber of all chambers. 800 μL of complete culture medium was added to the control group in the lower chamber. 800 μL of complete culture medium containing 25 μM, 30 μM, and 35 μM shantonin was added to the experimental groups, respectively. Each group was placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for 24 hours. All chambers were taken out, washed twice with PBS, and 4% tissue fixative was added to fix the cells at room temperature for 20 minutes. After discarding the fixative, wash twice with PBS, add crystal violet to stain the cells for 20 minutes, wash twice with PBS and discard the liquid. After air drying at room temperature, use the cell imaging system EVOS TM The M7000 imaging system captures the image.

[0039] 2.2 Experiment on scutellarin-induced autophagy in ATC cells

[0040] (1) Effects of scutellaria baicalensis on apoptosis inhibitors and necrosis inhibitors

[0041] Human anaplastic thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase were selected and the cell count of each cell was 2×10 3The number of cells was inoculated in a 96-well cell culture plate containing 100 μL complete medium in each well. After the cells adhered to the wall, the supernatant was discarded, and 100 μL of complete medium containing 10 μM necroptosis inhibitor Necrostatin-1 (Nec-1) and apoptosis inhibitor Z-VAD-FMK (Z-Vad) were added as drug pretreatment wells. After the 96-well plate was placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for 4 hours, the supernatant was discarded, and 200 μL of complete medium containing 0, 30, 40, and 50 μM shantonin were added to the corresponding small wells of the above drug pretreatment. In addition, 200 μL of complete medium with an equal volume of DMSO as 50 μM shantonin was set as a control group, and 200 μL of complete medium was added as a blank control group, with 3 replicates in each group. After the 96-well plate was placed in an incubator with 5% CO2 and a temperature of 37°C for 48 hours, 10 μL of CCK-8 reagent was added to each well and placed in an incubator with 5% CO2 and a temperature of 37°C for 2 hours; the absorbance value (A) of each well was measured using an ELISA reader at a wavelength of 450 nm. The survival rate of all cells at 48 hours was calculated, and the survival rate % = (A 药物组 -A 空白组 ) / (A 对照组 -A 空白组 )×100%, the experiment was repeated 3 times.

[0042] (2) Cell imaging experiments

[0043] Undifferentiated thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase were selected and the cells were counted and 2×10 5 Cells were inoculated in a 6-well plate. After the cells adhered to the wall, the original culture medium was discarded and 2000 μL of complete culture medium containing 50 μM shantonin was added. The control group was added with 2000 μL of complete culture medium. The 6-well plate was placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for 24 hours. The six-well plate was taken out and placed in the cell imaging system EVOS TM Imaging and shooting in the M7000 imaging system.

[0044] (3) Effects of shantonin on autophagy inhibitors

[0045] Human anaplastic thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase were selected and the cell count of each cell was 2×10 3The number of cells was inoculated in a 96-well cell culture plate containing 100 μL complete medium in each well. After the cells adhered to the wall, the supernatant was discarded and 100 μL of complete medium containing 10 μM autophagy inhibitor chloroquine (CQ) was added. After the 96-well plate was placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for 4 hours, the supernatant was discarded and 200 μL of complete medium containing 0, 30, 40, and 50 μM shantonin was added. In addition, a control group with an equal volume of DMSO complete medium of 200 μL and a blank control group with 200 μL complete medium was set up, with 3 replicates in each group. After the 96-well plate was placed in an incubator with 5% CO2 and a temperature of 37°C for 48 hours, 10 μL of CCK-8 reagent was added to each well and placed in an incubator with 5% CO2 and a temperature of 37°C for 2 hours; the absorbance value (A) of each well was measured using an ELISA reader at a wavelength of 450 nm. The survival rate of all cells at 48 hours was calculated, and the survival rate % = (A 药物组 -A 空白组 ) / (A 对照组 -A 空白组 )×100%, the experiment was repeated 3 times.

[0046] (4) Confocal experiment

[0047] Undifferentiated thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase were selected and the cells were counted and 2×10 4 Cells were inoculated in 4-well dishes, with a volume of 2000 μL per well. After the cells adhered to the wall, the culture medium was discarded, and 2000 μL of complete culture medium containing 50 μM shantonin was added to each well. 2000 μL of complete culture medium was added to the control group. The 4-well dishes were placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for 24 hours. The culture medium in the wells was discarded, and the cells in each well were fixed with 4% paraformaldehyde for 20 minutes. 2000 μL of complete culture medium containing 5% endoplasmic reticulum tracer, lysosome tracer and mitochondrial tracer were added respectively, and incubated in the dark for 8 hours. The supernatant was discarded, and the cells in the dish were washed with PBS, and placed in a STELLARISDIVE multiphoton microscope for imaging.

[0048] 2.3 Study on the effect of shantonin on autophagy-related proteins in ATC cells

[0049] Undifferentiated thyroid cancer cells 8505C and KMH-2 in the logarithmic growth phase were selected and the cells were counted and 2×10 5Each cell was inoculated in a 6-well plate with a volume of 2000 μL per well. After the cells adhered to the wall, the original culture medium was discarded, and 2000 μL of complete culture medium containing 0, 30, 35, 40, 45, and 50 μM of sophuronosyl ester was added to each well. 2000 μL of complete culture medium was added to the control group. The six-well plate was placed in an incubator with a 5% CO2 concentration and a temperature of 37°C for 24 hours. The six-well plate was taken out, the cells were digested with trypsin, and the cells were collected by centrifugation. After discarding the supernatant, the cells were lysed using Western IP lysis buffer, which contained 10% protease inhibitors. The cells were lysed on ice for 20 minutes and lysed twice with an ultrasonic disruptor at 25W for 10 seconds each time. The supernatant was extracted by centrifugation to obtain the protein, and the protein concentration was determined using a biuret protein assay kit. The protein was analyzed by FuturePAGE TM 4-20% 12-well gel was separated and then transferred to PVDF membrane by rapid transfer system. PVDF membrane was blocked with 5% skim milk powder for 2h at room temperature. All PVDF membranes were incubated with primary antibodies (p-mTOR, p-AKT473, p-AKT308, p-4EBP1, β-Actin, p-S6K S235 / 236, LC3B-Ⅰ, LC3B-Ⅱ) at 4°C for 8h, then washed with TBST and incubated with goat anti-rabbit IgG, goat anti-mouse IgG and monkey anti-goat antibodies at room temperature for 2h. The target bands were pretreated with FDbioDura ECL kit. Exposure observation was performed by ChemiDoc-MP imager.

[0050] 2.4 Statistical analysis

[0051] All quantitative data were statistically analyzed using Graphpad Prism 9.0 software, and multifactorial analysis of variance was used to compare the means among multiple groups. When the statistical value P < 0.05, the difference was statistically significant.

[0052] 3 Results and analysis

[0053] 3.1 Sophora flavescens inhibits the proliferation of anaplastic thyroid cancer cells

[0054] Figure 1 The growth of different anaplastic thyroid cancer cells after 48 hours of treatment with sauerkraut; A shows that sauerkraut has a significant inhibitory effect on the growth of anaplastic thyroid cancer cell lines, and the sensitivity of different cell lines is different. Among them, 8505C and KMH-2 cells are the most sensitive to sauerkraut, so these two cells are selected as the main research cells. At the same time, the scratch experiment (B) and the clone formation experiment (C) show that the cell growth rate is significantly slowed down under the treatment of sauerkraut. In the migration and invasion experiment (D), the results show that sauerkraut can significantly inhibit the migration ability of cancer cells, and the inhibitory effect increases with the increase of concentration.

[0055] 3.2 Sophora flavescens promotes cell autophagy

[0056] When cells were treated with apoptosis inhibitors Necrostatin-1 (Nec-1) and Z-VAD-FMK (Z-AD) and then treated with scutellaria baicalensis, it was found that cell survival rate increased and apoptosis was reversed ( Figure 2 (A).

[0057] Autophagy helps tumor cells form autophagosomes and enhances tumor cell apoptosis. After 24 hours of scutellaria baicalensis treatment, we found through microscopic imaging that cells produced autophagic vesicles ( Figure 2 In B). Figure 2 Based on the results in B, we used Chloroquine (CQ) autophagy inhibitor to pretreat cells and then added scutellaria baicalensis to treat them. The results showed that the cell survival rate was also improved to a certain extent ( Figure 2 (C). Subsequent endoplasmic reticulum and mitochondrial probes showed a significant increase in endoplasmic reticulum and mitochondria in the cells, indicating the occurrence of endoplasmic reticulum autophagy and mitochondrial autophagy. Lysosomal probes found that compared with the control group, a large amount of fluorescence intensity was generated in the cells, indicating that after scutellaria baicalensis treatment, the cells tended to form autophagosomes and use them to guide cell apoptosis ( Figure 2 (middle D).

[0058] 3.3 Effects of shantonin on autophagy-related proteins

[0059] mTOR is a serine / threonine kinase and a member of the PI3K-related kinase family. It can participate in the formation of mTORC1 and mTORC2. mTORC1 inactivates the autophagy regulatory complex formed by ULK1 and its interacting proteins through phosphorylation, thereby affecting the biogenesis of autophagosomes. LC3B, as a widely used autophagy marker, is involved in multiple processes of autophagosome formation. When autophagy is induced, LC3B is proteolytically cleaved by Atg4 to generate LC3-I. LC3-I is activated by Atg7 and then coupled with phosphatidylethanolamine (PE) in the membrane to generate processed LC3-II. The processed LC3-II is recruited to the growing phagosome.

[0060] In order to explore the role of shantonin as an inducer of autophagy in anaplastic thyroid cancer cells, the protein levels of p-mTOR, p-AKT308, p-AKT473, p-4EBP1, p-S6K, LC3B-Ⅰ and LC3B-Ⅱ were detected by Western Blot experiment. Figure 3). The results showed that phosphorylated AKT308, phosphorylated AKT473 and phosphorylated EBP1 proteins indicated that the cell's response to survival and growth signal transduction pathways were inhibited, proving that the cells gradually died. Phosphorylated mTOR was inhibited, inhibiting downstream phosphorylated S6K protein, unable to participate in downstream gene activation and regulation, and promoting cell autophagy. LC3B produced obvious cleavage forms and increased with increasing concentration, indicating that with increasing concentration, cells produced obvious autophagy. After the cells were treated with shantonin, the AKT / mTOR pathway-related proteins were downregulated and the LC3B cleavage form was produced. Shantonin induced autophagy through the AKT / mTOR pathway and caused tumor cell death through autophagy.

[0061] The above embodiments are only used to help understand the method of the present invention and its core concept. It should be noted that, for those skilled in the art, without departing from the principles of the present invention, several improvements and modifications may be made to the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A use of scutellaria baicalensis as an autophagy inducer for anaplastic thyroid cancer.

2. The use according to claim 1, characterized in that The inducer can promote autophagy of anaplastic thyroid cancer cells.

3. The use according to claim 2, characterized in that The inducer promotes autophagy of anaplastic thyroid cancer cells by inhibiting the AKT / mTOR pathway.

4. Use of sophora japonicum in the preparation of a drug for treating anaplastic thyroid cancer.

5. The use according to claim 4, characterized in that The drug can inhibit the proliferation of anaplastic thyroid cancer cells.

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