Use of endoperoxide in combination with glutamine deprivation in the preparation of iron death-sensitive acsl4-independent tumor cell metastasis inhibiting drugs

Abstract

The application provides an application of intracellular peroxide combined glutamine deprivation in an ACSL4-independent tumor cell metastasis inhibiting drug sensitive to ferroptosis. The technical problem in the prior art that ferroptosis tolerance is more conducive to tumor cell metastasis is solved, and a new intervention strategy for effectively inhibiting tumor cell metastasis with low ferroptosis sensitivity is developed.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to application of endogenous peroxide combined with glutamine deprivation in preparation of tumor cell metastasis inhibiting drugs sensitive to iron death independent of ACSL4. BACKGROUND

[0002] Iron death is a form of programmed cell death different from apoptosis and necrosis caused by the accumulation of iron ion-dependent lipid peroxidation, which is of great significance for the development of new tumor treatment strategies. Iron death and tumor metastasis, especially the process of epithelial-mesenchymal transition (EMT), are closely related to lipid metabolism.

[0003] Recent studies have shown that the accumulation of lipid peroxidation can promote tumor cell metastasis, and metastatic EMT cancer cells also become more sensitive to the iron death pathway. The intercellular interaction mediated by E-cadherin (E-cad) in epithelial cells can inhibit iron death by activating intracellular NF2 and Hippo signaling pathways, and inhibition of the signaling axis can promote iron death by up-regulating lipid peroxidation modulators through transcriptional coactivator YAP. When head and neck cancer cells are treated with the classic iron death inducer Rsl3, silencing the EMT marker ZEB1 reduces lipid peroxide production. In addition, phospholipids containing polyunsaturated fatty acids (PUFAs) (PUFA-PLs) are the main substrates of lipid peroxidation. Long-chain acyl-coenzyme A synthetase 4 (ACSL4) is a key component of membrane lipids and a key enzyme in the synthesis and modification of PUFA-PLs, which can induce cell sensitivity to iron death and increase cell membrane fluidity. Existing research believes that ACSL4 is an important pharmacological target for the treatment of iron death-related diseases, but there is currently no compound that directly targets ACSL4. The transformation process of EMT requires high levels of PUFA-PLs to increase the fluidity of the cell membrane, so this may also be the reason why EMT cancer cells become more sensitive to iron death.

[0004] Glutamine (Gln) is the most abundant amino acid in the human and animal circulation and in the body's amino acid pool, and it is also the most functional amino acid. Because the survival of cancer cells rapidly consumes exogenous Gln, leading to a lower content of Gln in tumor cells. Therefore, the rapid development of tumors depends on the reprogramming and utilization of Gln to maintain its growth. Gln deprivation can induce EMT and further metastasis of cancer cells by up-regulating the master regulator of EMT, Slug, through the MEK / ERK signal and ATF4 pathway. On the other hand, the metabolism of Gln is also closely related to the sensitivity of cells to ferroptosis. The core sensing signal pathway of amino acid metabolism is mTORC1, and mTORC1 is closely related to EMT and ferroptosis of cancer cells. In recent years, some documents have reported that the mutation of β-catenin in hepatocellular carcinoma leads to the overexpression of Wnt-specific targeting GLUL, which promotes the synthesis of Gln by activating the activity of glutamine synthetase (GS), and then activates mTORC1 and phosphorylates its downstream targets S6K1 and 4EBP1 to increase protein synthesis. Endoperoxides are a class of organic compounds containing a peroxide bridge. They contain a peroxide bridge (O-O) in their structure and exhibit a variety of properties and reactions in chemical reactions. Endoperoxides are mainly divided into 1,2-dioxane, 1,2,4-trioxane, 1,2,4,5-tetraoxane, 1,2-dioxolane and 1,2,4-trioxolane, etc. according to the size of the ring and the substitution position of the heteroatom. These endoperoxides have various biological activities, such as anti-malaria, anti-tumor, etc., and their physicochemical properties are relatively stable, which have attracted widespread attention.

[0005] Among them, artemisinin (ART) is a safe and reliable natural source compound containing a peroxide bridge group in its structure. In recent years, it has been found that ART can not only inhibit the progression of tumors, but also inhibit the metastasis of tumors by affecting EMT and angiogenesis. The effect of ART may also be related to the metabolism of Gln. Literature reports that inhibiting Gln hydrolysis enzyme GLS1 can not only block the metabolic reprogramming of hepatocellular carcinoma cells, but also reduce the dosage of dihydroartemisinin (DHA). FINO2, as an organic endoperoxide, has good anti-tumor effect.

[0006] Based on this, by studying the mechanism and characteristics of ferroptosis induced by endoperoxides and its influence on the EMT process of tumor cells caused by Gln deprivation, it is expected to provide a new intervention strategy for inhibiting the metastasis of tumor cells. SUMMARY

[0007] To this end, the technical problem to be solved by the present application is to provide the application of endoperoxide combined with glutamine deprivation in tumor cell metastasis inhibition drugs independent of ACSL4 iron death sensitivity. The technical problem of the prior art that iron death tolerance is more conducive to tumor cell metastasis is solved, and a new intervention strategy is developed for low iron death sensitivity, thereby effectively inhibiting tumor cell metastasis.

[0008] The present application provides a technical solution: the application of endoperoxide combined with glutamine deprivation in tumor cell metastasis inhibition drugs independent of ACSL4 iron death sensitivity, wherein the glutamine deprivation includes blocking glutamine supply and / or synthesis.

[0009] Preferably, an mTORC1 inhibitor is further included, and the endoperoxide combined with glutamine deprivation and the mTORC1 inhibitor inhibit the epithelial-mesenchymal transition of tumor cells.

[0010] Preferably, the epithelial-mesenchymal transition of tumor cells is inhibited by regulating the mTORC1 signaling pathway.

[0011] Preferably, the regulation of the mTORC1 signaling pathway is up-regulation of the expression of mTORC1 protein. Preferably, the endoperoxide is 1,2,4-trioxane endoperoxide.

[0012] Preferably, the 1,2,4-trioxane endoperoxide is artemisinin or FINO2.

[0013] Preferably, the tumor includes triple-negative breast tumor.

[0014] Preferably, the tumor cell metastasis inhibition drug contains an effective amount of artemisinin and / or FINO2 for inhibiting the epithelial-mesenchymal transition of tumor cells.

[0015] Beneficial effects:

[0016] The application of endoperoxide combined with glutamine deprivation in tumor cell metastasis inhibition drugs independent of ACSL4 iron death sensitivity is provided. The characteristics of endoperoxide-mediated iron death sensitivity under Gln deprivation conditions are first confirmed. Based on the differential expression of ACSL4 under different Gln supply conditions, it is found that endoperoxide can be used to interfere with the mechanism of Gln deprivation leading to resistance of tumor cells, especially TNBC cells, to classic iron death inducers and EMT.

[0017] The relationship between the lipid metabolism changes caused by Gln deprivation and EMT and ferroptosis tolerance is first confirmed. The role of Gln deprivation in regulating EMT and tolerance to classic ferroptosis inducers is confirmed; and the mechanism of mTORC1-mediated lipid metabolism changes in the role of Gln deprivation in inducing tolerance to classic ferroptosis inducers and promoting EMT is revealed. A new solution is provided for inhibiting tumor cell metastasis caused by the development of tumor cells in clinical glutamine starvation therapy or interference with glutamine metabolism.

[0018] In the method of the present application, 1,2,4-trioxane endoperoxides such as ART, FINO2 endoperoxides combined with glutamine deprivation and mTORC1 inhibitors achieve the effect of inhibiting tumor cell metastasis by inducing ferroptosis. Compared with classic ferroptosis inducers, the method of the present application clarifies that 1,2,4-trioxane endoperoxides such as ART, FINO2 combined with glutamine deprivation and mTORC1 inhibitors can be used for the treatment of breast cancer metastasis and clarify the mechanism thereof. The present application lays a foundation and provides a new idea for the development of effective drugs for inhibiting tumor cell metastasis, especially under the condition of Gln deprivation. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to make the content of the present application more easily understood, the present application will be further described in detail below according to specific embodiments of the present application and in conjunction with the accompanying drawings.

[0020] Figure 1 The technical path of the present application is shown in the schematic diagram;

[0021] Figure 2 The experimental results of Gln deprivation induced TNBC cells to be sensitive to ferroptosis induced by classic ferroptosis inducers in the present application are shown in the schematic diagram;

[0022] Figure 3 The experimental results of Gln deprivation induced TNBC cells to be sensitive to ferroptosis induced by FINO2 in the present application are shown in the schematic diagram;

[0023] Figure 4 The results of the characteristics of endoperoxides in TNBC cells induced by ferroptosis in Example 1 of the present application are shown in the schematic diagram;

[0024] Figure 5 The experimental results of the influence of endoperoxides on Gln sufficient mediated resistance to classic ferroptosis inducers independent of ACSL4 in Example 1 of the present application are shown in the schematic diagram A;

[0025] Figure 6 The experimental results of the influence of endoperoxides on Gln sufficient mediated sensitivity to FINO2 independent of ACSL4 in Example 1 of the present application are shown in the schematic diagram B;

[0026] Figure 7 Schematic diagram C shows the experimental results of the effect of peroxides on DNA sensitivity mediated by sufficient ACSL4-independent Gln in Example 1;

[0027] Figure 8 Schematic diagram D shows the experimental results of the effect of peroxides on resistance to classical ferroptosis inducers mediated by ACSL4-independent Gln deprivation in Example 1.

[0028] Figure 9 This is a schematic diagram (E) showing the experimental results of the effect of peroxides on FINO2 sensitivity mediated by ACSL4-independent Gln deprivation in Example 1.

[0029] Figure 10 This is a schematic diagram (F) showing the experimental results of the effect of peroxides on DHA sensitivity mediated by ACSL4-independent Gln deprivation in Example 1.

[0030] Figure 11 This is a schematic diagram (A) showing the experimental results of ferroptosis induced in ACSL- / -MEF cells by peroxide via iron oxide in Example 1 of this embodiment;

[0031] Figure 12 This is a schematic diagram (B) showing the experimental results of ferroptosis induced in ACSL- / -MEF cells by peroxide via iron oxide in Example 1 of this embodiment;

[0032] Figure 13 This is a schematic diagram (C) showing the experimental results of ferroptosis induced in ACSL- / -MEF cells by peroxide via iron oxide in Example 1 of this embodiment;

[0033] Figure 14 This is a schematic diagram (D) showing the experimental results of ferroptosis induced in ACSL- / -MEF cells by peroxide via iron oxide in Example 1 of this embodiment;

[0034] Figure 15 This is a schematic diagram (E) showing the experimental results of ferroptosis induced in ACSL- / -MEF cells by peroxide via iron oxide in Example 1 of this embodiment;

[0035] Figure 16 This is a schematic diagram of the experimental results of Gln deprivation promoting EMT in TNBC cells in Example 1 of this embodiment;

[0036] Figure 17 This is a schematic diagram of the cell migration experiment results of Gln-induced ferroptosis resistance in TNBC cells during EMT.

[0037] Figure 18 This is a schematic diagram (A) showing the experimental results of the potential mechanism of Gln deprivation on the metastasis of TNBC cells in Example 1 of this embodiment;

[0038] Figure 19 Figure B is a schematic diagram of experimental results of the potential mechanism of Gln deprivation for TNBC cell metastasis in Example 1;

[0039] Figure 20 Figure A is a schematic diagram of experimental results of mice, Figure B is a comparison of results of a control group, a single mTORC1 inhibitor group, a single ATS group, an ATS combined with an mTORC1 inhibitor group, and Figure C is a schematic diagram of results of tumor cell metastasis under the action of DHA combined with an mTORC1 inhibitor. DETAILED DESCRIPTION

[0040] The present application will be described in detail below with reference to the accompanying drawings and embodiments. The principles and characteristics of the present application are described below with reference to the accompanying drawings, and it should be noted that the embodiments and features in the embodiments in the present application can be combined with each other without conflict. The embodiments are only used to explain the present application and are not used to limit the scope of the present application.

[0041] The reagents used in the present specification are commercially available products unless otherwise specified.

[0042] The present application focuses on Gln deprivation inducing EMT in TNBC that is resistant to classic iron death inducers, and mainly studies 1,2,4-trioxane endoperoxides, especially ART and FINO2, which induce iron death that is not sensitive to lipid metabolism and effectively inhibit the mechanism of TNBC metastasis. The mechanism of Gln deprivation mediated tumor cell metastasis and resistance to classic iron death inducers is found and verified by Western blotting and CCK-8 and other methods; it is confirmed by animal metastasis models, lentivirus transfection and drug intervention techniques that endoperoxides may become an effective treatment for TNBC cells to resist oxidative stress in the blood and occur blood metastasis.

[0043] The research content of the present application includes the following four aspects:

[0044] (1) Study the effect of Gln deprivation on the EMT process and iron death sensitivity of TNBC

[0045] Based on the fact that Gln can regulate the accumulation of lipid peroxidation and EMT in tumor cells, first, we confirmed the effect of Gln deprivation on EMT, metastatic ability and ferroptosis sensitivity of breast cancer cells, and clarified that Gln deprivation can induce EMT in TNBC cells that is resistant to classic ferroptosis inducers. By comparing the differences in drug sensitivity of Gln-deprived cells to classic ferroptosis inducers (Rsl3 and Erastin) and internal peroxides (FINO2 and ART), and the EMT mediated by them, we clarified that internal peroxides can inhibit the EMT induced by Gln deprivation that is resistant to classic ferroptosis inducers.

[0046] (2) Study the process of lipid metabolism changes caused by Gln deprivation

[0047] Wnt-β-catenin-GS-Gln in hepatocellular carcinoma promotes cell metabolism, proliferation and survival through phosphorylation of mTORC1. The relationship between mTORC1 and lipid metabolism is complex. It can not only inhibit lipid peroxidation through SREBP1 / SCD1-mediated MUFA synthesis, but also promote lipid peroxidation accumulation by affecting the content of GSH, in addition, it can also promote cell metastasis through S6K1 / 4E-BP1. Verify that mTORC1-mediated MUFA is the key to Gln deprivation causing tumor cells to resist classic ferroptosis inducers by activating GS activity. Further study the EMT of breast cancer cells mediated by Gln deprivation through phosphorylation of mTORC1 substrates S6K1 and 4E-BP1. Clarify that mTORC1 is the key node of lipid metabolism reprogramming caused by Gln deprivation and mediates EMT and resistance to classic ferroptosis inducers in breast cancer.

[0048] (3) Study the mechanism and characteristics of internal peroxide-induced ferroptosis

[0049] ACSL4-mediated lipid metabolism is particularly critical in Rsl3-induced ferroptosis, while internal peroxides can directly oxidize PUFA-PLs. Therefore, first, we confirmed that internal peroxide-induced ferroptosis is independent of ACSL4. By comparing the expression of ACSL4 under different Gln supply conditions, we clarified that Gln deprivation-induced resistance to classic ferroptosis inducers in TNBC cells can be independent of ACSL4, and whether there is a significant relationship between Gln deprivation-mediated EMT and ACSL4, and finally confirmed that internal peroxides can induce ACSL4-independent ferroptosis and promote the killing of Gln deprivation-induced EMT cells.

[0050] (4) Study the effect of internal peroxide, especially ART, on TNBC metastasis and its mechanism

[0051] In view of the possible similarity of the mechanism of action of ART and FINO2, it is clear that ART can effectively interfere with Gln deprivation-mediated resistance of TBC cells to classic ferroptosis inducers and block Gln deprivation-mediated metastasis of TBC cells, and this process also does not depend on ACSL4. In addition, since ART can regulate tumor progression by inhibiting mTORC1, the present application will study whether ART can change the mTORC1-mediated lipid metabolism reprogramming. The in vivo metastasis of tumor cells can be effectively interfered by the endoperoxide represented by ART, and its mechanism of action is studied by verifying the in situ mouse breast cancer lung metastasis model. The technical approach adopted by the present application is shown in Figure 1 .

[0052] Embodiment 1

[0053] The present embodiment is the application of endoperoxide combined with glutamine deprivation in tumor cell metastasis inhibition drugs that are independent of ACSL4 ferroptosis sensitivity. Taking triple-negative breast cancer (TNBC) as an example, TNBC is a special type of breast cancer characterized by the absence of estrogen receptors (ER), progesterone receptors (PR), and human epidermal growth factor receptor 2 (HER2). This type of breast cancer accounts for about 10% to 20% of all breast cancer cases. Due to the lack of these receptors, triple-negative breast cancer is ineffective for hormone therapy and HER2-targeted therapy, and the treatment options are relatively limited. The incidence of triple-negative breast cancer is higher in young women and has a higher risk of invasion and recurrence. In addition, the prognosis of triple-negative breast cancer is relatively poor because of its fast growth rate and easy metastasis.

[0054] TNBC is ineffective for hormone therapy and HER2-targeted therapy, and the treatment options are relatively limited. In addition, TNBC has a relatively poor prognosis due to its fast growth rate and easy metastasis of cells, and existing technologies are difficult to effectively inhibit the metastasis of TNBC cells.

[0055] In the present embodiment, a new EMT phenomenon with ferroptosis resistance characteristics is found in TNBC, which is not consistent with the current conclusion that EMT cells are more sensitive to ferroptosis. The research process of the present embodiment shows that this ferroptosis resistance is more conducive to the cells undergoing EMT to withstand the oxidative stress environment in the blood after entering the blood, thereby occurring more effective metastasis. Therefore, the present application discloses that ferroptosis resistance is a key feature of the strong metastasis ability of TNBC.

[0056] Long-chain acyl-CoA synthetase 4 (ACSL4) is a key component of membrane lipids and a key enzyme in the synthesis and modification process of polyunsaturated fatty acids (PUFAs), which can induce cell sensitivity to ferroptosis and increase the fluidity of cell membranes.

[0057] Gln is more dependent on the rapid development of TNBC. This embodiment simulates the condition that Gln is severely consumed during the development of TNBC by removing Gln in the cell culture medium, and Gln is added to the culture medium as the Gln+ experimental group.

[0058] Referring to Figure 2 , it is shown that EMT of 4T1 cells and HCC1973 cells against iron death induced by inducer Erastin and EMT of MDA-MB-231 cells against iron death induced by inducer Rsl3 in Gln+ and Gln- conditions, both of which show that cells in the Gln- group are more sensitive to EMT against inducer-induced iron death. Referring to Figure 3 , Gln deprivation induces TNBC cells to be still sensitive to FINO2-induced iron death.

[0059] This embodiment determines whether Gln deprivation-induced TNBC cells to be resistant to classical iron death inducers can be independent of ACSL4 and whether Gln deprivation-mediated EMT has a significant relationship with ACSL4 by comparing the expression of ACSL4 under different Gln supply conditions. Specifically, 1) detect the protein expression of ACSL4 in Gln+ and Gln- group cells by Western blotting experiment; 2) detect the sensitivity of cells with knockdown or inhibition of ACSL4 to iron death inducers Rsl3, Erastin, FINO2, MLH, TBH, ART and DHA by Gln deprivation; 3) detect the migration ability of cells with knockdown or inhibition of ACSL4 after treatment with low concentration of endoperoxide. Referring to Figure 4 , by detecting the characteristics of endoperoxide-induced iron death in TNBC cells, the result of low expression of ACSL4 in Gln-deprived TNBC cells is obtained, and finally it is confirmed that endoperoxide can induce iron death independent of ACSL4.

[0060] Since ACSL4-mediated lipid oxidation pathway is considered to be an important process of lipid peroxide production in Rsl3-induced iron death caused by loss of reducing capacity, this embodiment also detects the role of ACSL4 in Rsl3, FINO2 and DHA-induced iron death, respectively, and finds that knockdown of ACSL4 has a greater impact on classical iron death inducer Rsl3, referring to Figure 5 ; but it does not affect the cytotoxicity produced by FINO2 and DHA, referring to Figure 6 and Figure 7 In addition, knockdown of ACSL4 in Gln-deprived cells shows slight resistance to Rsl3-induced iron death, and has no effect on the sensitivity of FINO2 and DHA-mediated iron death, referring to Figures 8-10The above results show that, given that the sensitivity to ferroptosis mediated by endogenous peroxides is independent of ACSL4, it can be used to treat cells that are induced by Gln deprivation to be resistant to classic ferroptosis inducers.

[0061] Further, in this embodiment, endogenous peroxides induce ferroptosis in TBC cells by oxidizing iron.

[0062] Referring to Figures 11-15 , the oxidative inducers FINO2, TBH, MLH, etc. can induce ferroptosis in cells by oxidizing iron, rather than other forms of cell death, and the process is independent of ACSL4.

[0063] Further, in this embodiment, endogenous peroxides inhibit the epithelial-mesenchymal transition (EMT) of TNBC. In this embodiment, the EMT of TNBC is inhibited by regulating the mTORC1 signaling pathway. Moreover, the mTORC1 signaling pathway is regulated by upregulating the expression of mTORC1 protein.

[0064] Based on the fact that Gln can regulate the accumulation of lipid peroxidation in tumor cells and EMT, first, the effect of Gln deprivation on the EMT, metastatic ability, and sensitivity to ferroptosis of breast cancer cells is verified, and it is determined that Gln deprivation can induce EMT in TBC cells that is resistant to classic ferroptosis inducers. By comparing the differences in drug sensitivity of Gln-deprived cells to classic ferroptosis inducers (Rsl3 and Erastin) and endogenous peroxides (FINO2 and ART) and the EMT mediated thereby, it is determined that endogenous peroxides can inhibit the EMT that is resistant to classic ferroptosis inducers caused by Gln deprivation.

[0065] To detect the mechanism of lipid metabolism changes in Gln deprivation in regulating tumor cell metastasis and resistance to ferroptosis, the following methods are used: 1) Western blotting is used to detect the content of GS and mTORC1 protein and the upstream and downstream relationship in Gln+ and Gln- groups; 2) Rapamycin (Rapa) is used to inhibit the mTORC1 signaling pathway, and the metastatic ability of Gln- group breast cancer cells and the sensitivity to ferroptosis inducers Rsl3, Erastin, FINO2, MLH, TBH, ART, and DHA are detected; the effect of rapamycin on Rsl3-induced ferroptosis after the addition of MUFA palmitoleic acid (16:1, PO) or oleic acid (18:1, OA) is detected; 3) after the phosphorylation of S6K1 and 4E-BP1 is inhibited by drugs, the E-cad / Snail protein expression and cell metastatic ability of Gln-deprived breast cancer cells are detected.

[0066] Referring to Figure 16, shows that Gln deprivation promotes the EMT process. The results show that the EMT phenomenon of 4T1 cells and MDA-MB-231 cells induced by ferroptosis occurs in the Gln+ and Gln- cases, the EMT marker E-cad is significantly reduced in the Gln- group of the two cell groups, and the EMT marker Snail is expressed in the Gln- group of the two cell groups. It can be seen that Gln deprivation can promote EMT of tumor cells resistant to classical ferroptosis inducers, which may be more conducive to EMT cells to withstand oxidative stress in the blood after entering the blood, thereby more effective metastasis. Unlike the prior art, EMT cancer cells that are metastatic also become more sensitive to the ferroptosis pathway.

[0067] Referring to Figure 17 , the migration experiment results show that the number of cells in the Gln- group without Lip-1 increases significantly, indicating that the number of migrating cells increases, i.e. glutamine deprivation induces EMT in TNBC.

[0068] To explore the possible mechanism of Gln deprivation promoting breast cancer metastasis, this embodiment also detects the expression of GS and mTORC1 protein after Gln deprivation, and the effect of mTORC1 inhibitor Rapa on EMT expression of breast cancer cells, referring to Figure 18 , shows that Gln deprivation can activate GS activity, accompanied by up-regulation of mTORC1 expression, referring to Figure 19 , shows that the mTORC1 signaling pathway can regulate EMT. The above results show that Gln deprivation may regulate breast cancer cell metastasis and resistance to classical ferroptosis inducers through mTORC1-mediated lipid metabolism changes.

[0069] In this embodiment, referring to Figure 20 , in vivo experiments detect the effect of ART on TNBC metastasis under Gln deprivation. The luciferase-labeled Control group (control group), Rapamycin group (mTORC1 inhibitor alone group), ATS group (internal peroxide alone group), and ATS+Rapamycin group (internal peroxide and mTORC1 inhibitor group) are used. Intraperitoneal injection of 5 mg / kg of drug treatment once a day, observe the metastasis of breast tumor to the lung in vivo by live animal fluorescence imaging technology, and observe the size and number of tumor nodules metastasized to the lung after dissection. In this embodiment, since the derivative ATS of ART has better stability, and the therapeutic effects of the two in tumors are similar, the in vivo experiment of ART drug treatment uses the derivative ATS of ART as the treatment drug. Referring to Figure 20The results show that, under Gln deprivation, the tumor signals of mice injected with ATS drugs alone and mice applied with mTORC1 inhibitors alone are significantly lower than those of the control group of mice without any drug injection; the number of lung metastatic nodules is significantly less; while the mice injected with ATS and mTORC1 inhibitor drugs at the same time show the lowest tumor signal and the least number of lung metastatic nodules.

[0070] It can be seen that ART, FINO2, which are representative of internal peroxides, combined with Gln deprivation and mTORC1 inhibitors are expected to develop into effective drugs for treating tumor cell metastasis.

[0071] Obviously, the above examples are only examples for the sake of clarity, and are not a limitation on the embodiments. Based on the above description, other different forms of changes or variations can also be made by those of ordinary skill in the art. Here, it is not necessary and impossible to exhaust all embodiments. The obvious changes or variations derived therefrom are still within the protection scope of the present invention.

Claims

1. Use of an inner peroxide in combination with glutamine deprivation, mTORC1 inhibitor for the preparation of a drug for inhibiting metastasis of tumor cells sensitive to ferroptosis independent of ACSL4, characterized in that, The mTORC1 inhibitor is rapamycin, the glutamine deprivation includes blocking glutamine supply and / or synthesis; the tumor is triple negative breast tumor; the endogenous peroxide is artemisinin or FINO2.

2. Use according to claim 1, characterized in that, The endogenous peroxide in combination with glutamine deprivation and mTORC1 inhibitor inhibits epithelial-mesenchymal transition of tumor cells.

3. Use according to claim 2, characterized in that, The epithelial-mesenchymal transition of tumor cells is inhibited by regulating mTORC1 signaling pathway.

4. Use according to claim 3, characterized in that, The regulation of mTORC1 signaling pathway is up-regulating the expression of mTORC1 protein.

5. Use according to claim 4, characterized in that, The tumor cell metastasis inhibiting drug contains an effective amount of artemisinin or FINO2 which inhibits epithelial-mesenchymal transition of tumor cells.

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