Application of luteolin in sensitizing cisplatin in the preparation of drugs for the treatment of esophageal squamous cell carcinoma

Abstract

The present invention relates to the use of luteolin to enhance the sensitization of cisplatin in the preparation of a drug for the treatment of esophageal squamous cell carcinoma. The present invention finds that luteolin can block the progression of esophageal squamous cell carcinoma cells in multiple aspects, including apoptosis induction, growth inhibition, and invasion inhibition. Luteolin can also significantly enhance the ability of cisplatin to inhibit the lymphatic metastasis of esophageal squamous cell carcinoma cells in a mouse model, demonstrating a significant synergistic effect between the two.

Description

Technical Field

[0001] The present invention relates to a combination drug, and in particular to the use of luteolin to enhance the sensitization of cisplatin in the preparation of a drug for treating esophageal squamous cell carcinoma. Background Art

[0002] Esophageal cancer is a malignant tumor of the digestive tract with high morbidity and mortality. Esophageal squamous cell carcinoma (ESCC) is usually diagnosed with advanced disease or metastasis, and cannot be treated with radical surgery. The currently recommended standard first-line treatment for this advanced or metastatic disease is chemotherapy, including fluoropyrimidine- and platinum-based treatment regimens. Adjuvant or neoadjuvant therapy based on cisplatin has become the standard treatment for advanced esophageal cancer. Cisplatin alone is prone to drug resistance and is the main cause of chemotherapy failure. The reason is related to the activation of multiple key cancer-promoting molecules and the signaling pathways they regulate. Therefore, based on drug resistance considerations, the combination of drugs is an important strategy to combat drug resistance.

[0003] In recent years, traditional Chinese medicine has been widely used by researchers in combination with other drugs to enhance synergy and reduce toxicity due to its comprehensive effects of multiple components, multiple pathways, and multiple targets. Some traditional Chinese medicines also have anti-tumor effects. Therefore, the research on the anti-tumor effect of traditional Chinese medicine as an auxiliary chemotherapy drug has become a current hot topic.

[0004] Cynoside (Cy), also known as luteolin-7-O-glucoside, is a flavonoid glycoside compound widely found in plants, mainly found in plants such as honeysuckle, snow chrysanthemum, Chinese lantern, and celery. It has antioxidant, antitussive, anti-inflammatory, liver-protective, and anti-tumor activities. The structural formula of cynoside is as follows:

[0005]

[0006] However, whether luteolin combined with cisplatin can enhance the effect of cisplatin against esophageal squamous cell carcinoma still needs further study. Summary of the Invention

[0007] The present invention found that the combination of luteolin and cisplatin can well enhance the anti-esophageal squamous cell carcinoma effect of cisplatin, and the combination shows an unexpected synergistic effect.

[0008] Therefore, in a first aspect, the present invention provides a pharmaceutical composition comprising luteolin or a pharmaceutically acceptable salt, prodrug, metabolite thereof and cisplatin.

[0009] In addition, the present invention also provides a kit comprising luteolin or a pharmaceutically acceptable salt, prodrug, metabolite thereof and cisplatin.

[0010] In one embodiment, the ratio of luteolin or a pharmaceutically acceptable salt, prodrug, or metabolite thereof to cisplatin is 0.5-60:1, based on the mass of luteolin and cisplatin.

[0011] Preferably, the ratio of luteolin or its pharmaceutically acceptable salt, prodrug, metabolite to cisplatin, based on the mass of luteolin and cisplatin, is 0.5:1, 1:1, 1.5:1, 3:1, 10:1, 20:1, 30:1, 35:1, 40:1, 50:1, 60:1, preferably, it is 1-40:1, or 1.5-35:1, most preferably, it is 1.5-3:1, or 35:1.

[0012] In a second aspect, the present invention also provides use of the pharmaceutical composition of the present invention in preparing a drug for treating esophageal squamous cell carcinoma.

[0013] In the present invention, the treatment of esophageal squamous cell carcinoma includes at least one of inhibiting the growth of esophageal squamous cell carcinoma cells, inhibiting the invasion of esophageal squamous cell carcinoma cells, inducing apoptosis of esophageal squamous cell carcinoma cells, and inhibiting the metastasis of esophageal squamous cell carcinoma cells.

[0014] Preferably, the metastasis includes lymphatic metastasis.

[0015] In one embodiment, the esophageal squamous cell carcinoma cells include esophageal squamous cell carcinoma cells KYSE410, esophageal squamous cell carcinoma cells KYSE450, and esophageal squamous cell carcinoma cells KYSE510.

[0016] In a third aspect, the present invention also provides a use of luteolin or a pharmaceutically acceptable salt, prodrug, or metabolite thereof in enhancing the anti-esophageal squamous cell carcinoma effect of cisplatin.

[0017] The present invention also provides a use of luteolin or a pharmaceutically acceptable salt prodrug or metabolite thereof in the preparation of a drug that enhances the anti-esophageal squamous cell carcinoma effect of cisplatin.

[0018] In the present invention, the pharmaceutically acceptable salt of luteolin includes acid addition salt or base addition salt of luteolin. In one embodiment, the pharmaceutically acceptable salt of luteolin is hydrochloride, hydrobromide, sulfate, phosphate, metaphosphate, acetate, propionate, hexanoate, cyclopentanepropionate, glycolate, pyruvate, lactate, malonate, succinate, malate, L-malate, maleate, oxalate, fumarate, trifluoroacetate, tartrate, L-tartrate, citrate, benzoate, 3-(4-hydroxybenzoyl)benzoate, cinnamate , mandelate, methanesulfonate, ethanesulfonate, 1,2-ethanedisulfonate, 2-hydroxyethanesulfonate, benzenesulfonate, toluenesulfonate, 2-naphthalenesulfonate, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylate, glucoheptonic acid, 3-phenylpropionic acid, pivalate, t-butylacetate, lauryl sulfate, gluconate, glutamate, hydroxynaphthoate, salicylate, stearate, muconate, butyrate, phenylacetate, phenylbutyrate, or valproate.

[0019] In the present invention, the prodrug refers to a derivative that can directly or indirectly provide luteolin after being administered to a patient, such as an ester of tagetoidin.

[0020] In the present invention, the metabolite refers to a pharmaceutically acceptable metabolic derivative of luteolin or a pharmaceutically acceptable salt thereof.

[0021] Preferably, the medicine or pharmaceutical composition of the present invention further comprises a pharmaceutically acceptable carrier.

[0022] Preferably, the luteolin or its pharmaceutically acceptable salt, prodrug, or metabolite and cisplatin in the pharmaceutical composition of the present invention can be administered in the same or different pharmaceutical formulations. The pharmaceutical dosage forms of luteolin or its pharmaceutically acceptable salt, prodrug, or metabolite and cisplatin can be the same or different. Luteolin or its pharmaceutically acceptable salt, prodrug, or metabolite and cisplatin can be administered simultaneously or sequentially.

[0023] In the medical uses described above, the administration time, number of administrations, and frequency of administration of luteolin or its pharmaceutically acceptable salts, prodrugs, metabolites, and cisplatin, etc., need to be determined according to the specific diagnosis results of the disease, which is within the technical scope mastered by those skilled in the art.

[0024] For example, when a treatment regimen for mice or rats is applied to humans, the effective doses of all drugs for humans can be converted by the effective doses of the drugs for mice or rats, which is also easy to achieve for ordinary technicians in this field.

[0025] As used herein, "synergy," "synergistic action," "synergistic effect," and the like are used to mean that the result of the combination of two compounds, components, or targeting agents is greater than the sum of the sum of each compound, component, or targeting agent alone. This improvement in the disease, condition, or disorder being treated is a "synergistic" effect.

[0026] As used herein, "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0027] The pharmaceutically acceptable carrier may comprise any conventional pharmaceutical carrier or excipient. The choice of carrier and / or excipient will largely depend on factors such as the particular mode of administration, the effect of the carrier or excipient on solubility and stability, and the nature of the dosage form.

[0028] Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents (such as hydrates and solvates). If necessary, pharmaceutical compositions can contain additional ingredients, such as flavorings, adhesives, excipients, etc. Therefore, for oral administration, tablets containing various excipients (such as citric acid) can be used with various disintegrants (such as starch, alginic acid and some composite silicates) and with adhesives (such as sucrose, gelatin and gum arabic). The example of excipient includes calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols without limitation. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc can often be used for tableting purposes. Similar types of solid compositions can also be used in soft and hard-filled gelatin capsules. Therefore, the non-limiting example of material includes lactose and high molecular weight polyethylene glycol. When aqueous suspensions or elixirs are desired for oral administration, the active compound therein may be combined with various sweetening or flavoring agents, colorings or dyes and, if desired, emulsifying or suspending agents and diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.

[0029] In the present invention, the drug or pharmaceutical composition can be, for example, in a form suitable for oral administration (as tablets, capsules, pills, powders, solutions, suspensions), a form suitable for parenteral injection (as sterile solutions, suspensions or emulsions), a form suitable for topical administration (as ointments or creams) or a form suitable for rectal administration (as suppositories).

[0030] In the present invention, the drug or pharmaceutical composition may be in a unit dosage form suitable for single administration of a precise dosage.

[0031] The medicament or pharmaceutical composition of the present invention can be administered orally. Oral administration can include swallowing, whereby the compound enters the gastrointestinal tract; or buccal or sublingual administration can be employed, whereby the compound enters the bloodstream directly from the mouth.

[0032] Formulations suitable for oral administration include solid preparations such as tablets; capsules containing microparticles, liquids, or powders; lozenges; chewable tablets; multiparticulates and nanoparticulates; gels; solid solutions; liposomes; films; ovoids; sprays, and liquid preparations. Liquid preparations include suspensions, solutions, syrups, and elixirs. Such preparations can be used as fillers in soft or hard capsules and typically include a carrier, for example, water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or a suitable oil, and one or more emulsifiers and / or suspending agents. Liquid preparations can also be prepared by reconstituting a solid.

[0033] For tablet dosage forms, depending on the dosage, the drug can account for 1% to 80% by weight of the dosage form, more typically 5% to 60% by weight of the dosage form. In addition to the drug, tablets generally contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinyl pyrrolidone, methylcellulose, microcrystalline cellulose, low alkyl substituted hydroxypropyl cellulose, starch, pregelatinized starch and sodium alginate. Typically, the disintegrant accounts for 1% to 25% by weight of the dosage form, preferably 5% to 20% by weight.

[0034] Binders are generally used to impart cohesiveness to tablet formulations. Suitable binders include microcrystalline cellulose, gelatin, sugar, polyethylene glycol, natural and synthetic gums, polyvinyl pyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methylcellulose. Tablets can also contain diluents, such as lactose (monohydrate, spray-dried monohydrate, anhydrous lactose etc.), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch and calcium hydrogen phosphate dihydrate.

[0035] The tablets may also optionally include surfactants, such as sodium lauryl sulfate and polysorbate 80, and glidants, such as silicon dioxide and talc. When present, the amount of surfactant is typically 0.2% to 5% by weight of the tablet, and the amount of glidant is typically 0.2% to 1% by weight of the tablet.

[0036] Tablets generally also contain lubricants such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and a mixture of magnesium stearate and sodium lauryl sulfate. Lubricants are generally present in an amount of 0.25% to 10% by weight, preferably 0.5% to 3% by weight of the tablet.

[0037] Other conventional ingredients include antioxidants, colorants, flavorings, preservatives and taste-masking agents.

[0038] Exemplary tablets contain up to about 80% by weight of the drug, about 10% to about 90% by weight of the binder, about 0% to about 85% by weight of the diluent, about 2% to about 10% by weight of the disintegrant, and about 0.25% to about 10% by weight of the lubricant.

[0039] Tablet blends can be compressed directly or by rollers to form tablets. Alternatively, tablet blends or portions of blends can be wet, dry, or melt granulated, melt-congealed, or extruded before tableting. The final formulation can include one or more layers and can be coated or uncoated; or encapsulated.

[0040] Solid dosage forms for oral administration can be formulated to be immediate and / or modified release. Modified release formulations include delayed, sustained pulsed, controlled, targeted, and programmed release.

[0041] The medicament or pharmaceutical composition of the present invention can also be administered directly into the bloodstream, muscle or internal organs. Suitable means of parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle) injectors, needle-free injectors and infusion techniques.

[0042] The medicament or pharmaceutical composition of the present invention can also be applied topically to the skin or mucous membrane, that is, transdermally or transdermally. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibers, bandages, and microemulsions. Liposomes can also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerol, polyethylene glycol, and propylene glycol.

[0043] The medicament or pharmaceutical composition of the present invention may also be administered intranasally or by inhalation, typically in dry powder form from a dry powder inhaler, or as an aerosol spray from a pressurized container, pump, sprayer, nebulizer or nebulizer.

[0044] Beneficial effects:

[0045] The present invention provides the use of luteolin to enhance cisplatin sensitivity in the preparation of a drug for the treatment of esophageal squamous cell carcinoma. Luteolin can block the progression of esophageal squamous cell carcinoma cells in multiple ways, including apoptosis induction, growth inhibition, and invasion inhibition. MTS results showed that luteolin enhanced the growth of cisplatin-resistant KYSE450 cells. Transwell assay results showed that luteolin enhanced the cisplatin-resistant KYSE450 cell invasion. Luteolin enhanced the apoptosis-inducing effect of cisplatin in KYSE450 cells and promoted PARP activation. In a KYSE450 tumor-bearing mouse model, luteolin combined with cisplatin effectively inhibited tumor cell volume growth, suppressed the expression of multiple tumor markers, promoted tumor cell apoptosis, and enhanced the anticancer effect of cisplatin. Luteolin also significantly enhanced the cisplatin-resistant lymphatic metastasis of the esophageal squamous cell carcinoma cell line KYSE450 in the mouse model. Luteolin and cisplatin exhibit significant synergistic effects in the treatment of esophageal squamous cell carcinoma. Their combined use can effectively improve anti-tumor efficacy, reduce drug dosage, minimize toxic side effects, and reduce the development of drug resistance. BRIEF DESCRIPTION OF THE DRAWINGS Description of the drawings:

[0047] Figure 1 This is a schematic diagram showing the results that luteolin in Example 1 can enhance the growth inhibition of esophageal squamous cell carcinoma cell lines by cisplatin.

[0048] Figure 2 This is a schematic diagram showing the results of Example 2 showing that luteolin can enhance the effect of cisplatin in inhibiting the invasion of esophageal squamous cell carcinoma cell lines.

[0049] Figure 3 This is a schematic diagram of the results of Example 3 showing that luteolin enhances cisplatin-induced apoptosis of esophageal squamous cell carcinoma cell lines and upregulation of cleaved PARP expression.

[0050] Figure 4 This is a schematic diagram of the results of Example 4 showing that luteolin enhances the effect of cisplatin in inhibiting the growth of KYSE450 tumors and the expression of multiple cancer-promoting markers in a xenogeneic tumor-bearing mouse model; and significantly upregulates the induction effect of cisplatin on the expression of cleaved PARP in KYSE450 tissues.

[0051] Figure 5 This is a schematic diagram of the results of Example 5 in which luteolin enhances cisplatin's ability to inhibit the lymphatic metastasis of KYSE450 tumors in a xenogeneic tumor-bearing mouse model.

[0052] Figure 6 The luteolin in Example 6 can achieve a more effective anti-cancer effect of sensitizing cisplatin in esophageal squamous cell carcinoma cells than in other types of tumor cells.

[0053] In the above figures, ** represents P < 0.01; *** represents P < 0.001. DETAILED DESCRIPTION

[0054] The following describes a preferred embodiment of the invention in detail. The examples are provided to better illustrate the invention and are not intended to limit the invention to the examples. Non-essential improvements and adjustments to the embodiments based on the invention remain within the scope of the invention.

[0055] The experimental methods in the following examples are all conventional methods unless otherwise specified. If no specific techniques or conditions are specified in the examples, they were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. Specific implementation method:

[0057] Example 1: Luteolin enhances the growth inhibitory effect of cisplatin on esophageal squamous cell carcinoma cell lines

[0058] Cell growth ability assay

[0059] KYSE450 cells were cultured at 3 × 10 3 Cells were seeded at a density of 100 cells / well in a 96-well plate. After the cells adhered to the wall, luteolin (10 μM) and different concentrations of cisplatin were added, used alone or in combination. After 72 hours, 10% MTS solution was prepared with RPMI 1640 medium, the cultured cells were removed, the upper medium was discarded, and the prepared MTS was added and incubated for 2 hours. The absorbance value was measured at 490 nm using a microplate reader. The growth inhibition rate of different concentrations of cisplatin was compared with the control group and input into GraphPadPrism software, and IC was analyzed using the "Dose-response inhibition" model. 50 The fitting value.

[0060] Compare with Figure 1 , the results showed that luteolin could enhance the growth inhibition of esophageal squamous cell carcinoma cell line by cisplatin.

[0061] Example 2: Luteolin enhances the inhibitory effect of cisplatin on esophageal squamous cell carcinoma cell line invasion

[0062] Cell invasion ability assay

[0063] KYSE450 cells were cultured in serum-free medium for 24 hours. Using an 8μm pore transwell, 100μL of Matrigel was added to the upper chamber of the transwell and incubated at 37°C for 1 hour to allow the Matrigel to solidify. The prepared KYSE450 cells were seeded into the upper chamber of the transwell. 800μL of RPMI 1640 medium containing 20% ​​fetal bovine serum was added to the lower chamber, along with luteolin (10μM) and cisplatin (10μM), either alone or in combination. The transwell was then placed in an incubator and incubated for 24 hours. After removal, the inner wall of the upper chamber was cleaned with a cotton swab. The invading cells in the lower chamber were isolated using dissociation solution, and a cell dye was added. The invasion rate was calculated using a microplate reader.

[0064] Compare with Figure 2 , the results showed that luteolin could enhance the cisplatin-inhibitory effect on the invasion of esophageal squamous cell carcinoma cell lines.

[0065] Example 3: Luteolin enhances the apoptosis-promoting effect of cisplatin on esophageal squamous cell carcinoma cell lines

[0066] 1. Determination of Cell Apoptosis Index

[0067] KYSE450 cells were seeded in RPMI 1640 medium supplemented with 10% fetal bovine serum. After cell attachment, luteolin (10 μM) and cisplatin (5 μM) were added, either alone or in combination. After 24 hours of incubation, the supernatant was collected by centrifugation. The supernatant was added to the wells of a 96-well apoptosis detection kit, sealed with a blocking membrane, and incubated at room temperature for 1 hour. The sample was removed, the 96-well plate was washed repeatedly, and the substrate solution (100 μL / well) was added. After shaking for 15 minutes, the 96-well plate was gently tapped, and the absorbance was measured at 405 nm.

[0068] 2. Detection of key apoptosis proteins in esophageal squamous cell carcinoma cell lines

[0069] KYSE450 cells were seeded in RPMI 1640 medium supplemented with 10% fetal bovine serum. After cell attachment, luteolin (10 μM) and cisplatin (5 μM) were added, either alone or in combination. After 24 hours of incubation, the cells were harvested along with the supernatant and centrifuged at 12,000 rpm for 10 minutes at 4°C. The precipitate was then collected. RIPA lysis buffer was added and the cells were lysed on ice for 1 hour, with shaking every 10 minutes. After lysis, the supernatant was collected after centrifugation at 12,000 rpm for 20 minutes at 4°C to obtain the protein lysate. The expression of cleaved poly (ADP-ribose) polymerase (PARP) was further analyzed using a high-throughput enzyme-linked immunosorbent assay (ELISA). PARP is a key apoptosis protein, and the expression of cleaved PARP can clearly measure the degree of cell apoptosis. Samples (100 μL / well) were sequentially added to a 96-well cleaved PARP ELISA plate and incubated at 37°C. After incubation, the 96-well plate was repeatedly washed. The cleaved PARP antibody was added to the 96-well plate and incubated at room temperature for 1 hour, then repeatedly washed and discarded. The reaction substrate was added to each well and incubated at room temperature in the dark. The reaction stop solution was then added and the absorbance was measured at 450 nm using a microplate reader.

[0070] Compare with Figure 3 The results showed that luteolin enhanced cisplatin-induced apoptosis of esophageal squamous cell carcinoma cell lines and upregulation of cleavage-type PARP expression.

[0071] Example 4: Luteolin enhances the anti-tumor growth and apoptosis-promoting effects of cisplatin on esophageal squamous cell carcinoma cell line KYSE450 in mice

[0072] KYSE450 cells were injected into the subcutaneous tissue of the upper limbs of nude mice (4-week-old female BALB / c-nu nude mice (average weight 14-15 g), purchased from Vital River Company) until the tumor grew to about 100 mm. 3 After about 3 weeks, the KYSE450 tumor-bearing mice were randomly divided into four groups: control group, luteolin group, cisplatin group and luteolin plus cisplatin group. The luteolin group (25 mg / kg) was administered orally once a day, the cisplatin group (5 mg / kg) was intraperitoneally injected once a week, and the luteolin plus cisplatin group was intraperitoneally injected with cisplatin (5 mg / kg, intraperitoneally injected once a week) on the basis of luteolin (25 mg / kg, administered orally, once a day) for a total of 3 weeks. The control group was given the same dose of normal saline by gavage every day for 3 consecutive weeks. The formula for calculating tumor volume is: Volume = Tumor Long Diameter × Tumor Transverse Diameter 2 ×0.5. Tumor tissues were collected on day 28, and protein lysates were prepared. ELISA was performed to observe the expression of a key proliferation protein (Ki67), an angiogenesis marker (CD31), and a key apoptosis protein (cleaved PARP) in KYSE450 tumors.

[0073] Compare with Figure 4 The results showed that luteolin enhanced the effect of cisplatin in inhibiting the growth of KYSE450 tumors and the expression of multiple cancer-promoting markers in a xenograft-bearing mouse model; and significantly upregulated the induction effect of cisplatin on the expression of cleaved PARP in KYSE450 tissues.

[0074] The Jin Zhengjun Q value method was used to evaluate the synergistic effect of the combination of luteolin and cisplatin in inhibiting tumor growth. The Q value was calculated using the following formula: Q = E a+b / (E a +E b -E a ×E b ). E a+b is the inhibition rate of the drug combination, E a and E b is the inhibition rate of drug treatment when administered alone. Q < 0.85 indicates antagonism, 0.85 ≤ Q < 1.15 indicates additive effect, and Q ≥ 1.15 indicates synergism.

[0075] After calculation, the Q value for tumor volume was 1.34; for Ki67, the Q value was 1.51; for CD31, the Q value was 1.39; and for cleaved PARP, the Q value was 1.44.

[0076] Example 5: Luteolin enhances the anti-tumor invasion promoting effect of cisplatin on esophageal squamous cell carcinoma cell line KYSE450 in mice

[0077] KYSE450 cells were inoculated into the footpads of nude mice (4-week-old female BALB / c-nu nude mice (average weight 14-15 g), purchased from Vital River). Approximately one week later, luteolin (25 mg / kg / day) and / or cisplatin (5 mg / kg / week) were administered via gavage. The experiment lasted for 5 weeks. Popliteal lymph nodes were collected, and lymphatic volume was calculated. Volume calculation formula: Volume = Long diameter × Transverse diameter 2 ×0.5.

[0078] Compare with Figure 5 The results showed that luteolin enhanced the effect of cisplatin in inhibiting the lymphatic metastasis of KYSE450 tumors in a xenograft-bearing mouse model.

[0079] Similarly, the Jin Zhengjun Q value judgment method was used to evaluate the synergistic effect of the combination of luteolin and cisplatin in inhibiting tumor lymphatic metastasis, and the Q value was 1.16.

[0080] Example 6: Luteolin can achieve a more effective anti-cancer effect of sensitizing cisplatin in esophageal squamous cell carcinoma cells than other types of tumor cells

[0081] Cell growth ability assay:

[0082] KYSE450 cells and the nasopharyngeal carcinoma cell line C666-1 were seeded in 96-well plates. After cell attachment, luteolin (10 μM) and various concentrations of cisplatin were added, either alone or in combination. After 72 hours, a 10% MTS solution was prepared in RPMI 1640 medium. The cultured cells were removed, the supernatant medium discarded, and the prepared MTS solution was added for incubation for 2 hours. Absorbance was measured at 490 nm using a microplate reader.

[0083] Compare with Figure 6 The results showed that luteolin could achieve a more effective anti-cancer effect of sensitizing cisplatin in esophageal squamous cell carcinoma cells than other types of tumor cells.

[0084] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. Use of a pharmaceutical composition in the preparation of a drug for treating esophageal squamous cell carcinoma, the pharmaceutical composition comprising luteolin or a pharmaceutically acceptable salt thereof and cisplatin, wherein the ratio of luteolin or a pharmaceutically acceptable salt thereof to cisplatin, based on the mass of luteolin and cisplatin, is 35:

1.

2. The use according to claim 1, characterized in that The treatment of esophageal squamous cell carcinoma includes at least one of inhibiting the growth of esophageal squamous cell carcinoma cells, inhibiting the invasion of esophageal squamous cell carcinoma cells, inducing apoptosis of esophageal squamous cell carcinoma cells, and inhibiting lymphatic metastasis of esophageal squamous cell carcinoma cells.

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