Application of madder extract in preparation of medicine for treating tumors

Abstract

The invention provides application of a madder extract in preparation of a medicine for treating tumors, and relates to the technical field of traditional Chinese medicines. The rubia cordifolia extract is prepared through a water extraction and alcohol precipitation method, and the specific steps comprise soaking, condensation reflux decoction twice, filtering, ethanol precipitation, standing, drying and the like. The rubia cordifolia extract disclosed by the invention can be used for remarkably inhibiting prostate cancer cell proliferation, inducing cell apoptosis and inhibiting metastasis, and has good targeting property and low toxic and side effects. Compared with a traditional treatment medicine, the rubia cordifolia extract disclosed by the invention, as a natural medicine, shows remarkable advantages in the aspects of safety, tolerance and treatment effect, and provides a new potential choice for treatment of urinary system tumors.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine, and in particular to application of a madder extract in preparing medicines for treating tumors. Background Art

[0002] Malignant tumors refer to tumors with abnormal cell growth and division, invasiveness and metastasis capabilities. They are usually characterized by rapid cell division and structural abnormalities, and can invade adjacent tissues and metastasize to distant organs. Malignant tumors can be divided into many types according to the location of occurrence, such as lung cancer, gastric cancer, breast cancer, colorectal cancer, liver cancer, esophageal cancer, lymphoma and urinary system tumors. There are many types of malignant tumors, and the pathogenic mechanism, treatment methods and drugs used for each tumor are significantly different.

[0003] Urinary system tumors are malignant tumors that occur in the urinary system (such as the kidneys, bladder, prostate, etc.), including kidney tumors, ureter tumors, bladder tumors, prostate tumors, etc.

[0004] Renal cell carcinoma (RCC) is the most common malignant tumor of the kidney, usually presenting with symptoms such as painless hematuria, low back pain, and masses. Renal pelvic cancer occurs at the junction of the ureters of the kidney, and a common symptom is hematuria. Tubulointerstitial tumors: relatively rare, often requiring imaging examinations and pathological diagnosis. Ureteral tumors are relatively rare, mainly urothelial carcinoma (i.e. urothelial carcinoma), often presenting with hematuria, and a small number of patients may have symptoms such as low back pain and abnormal urination. Bladder cancer is one of the most common tumors of the urinary system, usually urothelial carcinoma. The main symptoms include painless hematuria, frequent urination, urgency, and pain when urinating.

[0005] Prostate cancer is one of the most common malignant tumors in men, usually occurring in the peripheral area of ​​the prostate. The prostate is part of the male reproductive system, located below the bladder and in front of the rectum. Its main function is to secrete prostatic fluid, which is involved in the composition of semen. The occurrence of prostate cancer is related to many factors, including age, family history, diet, hormone levels, etc.

[0006] Prostate cancer is usually adenocarcinoma, and its specific types can be divided into peripheral prostate cancer, central prostate cancer, and transition zone prostate cancer. Peripheral prostate cancer is the most common, accounting for about 70% of cases. It is usually located in the peripheral area of ​​the prostate. Central and transition zone prostate cancers are relatively rare. Early prostate cancer usually has no obvious symptoms, so many patients are diagnosed without symptoms. As the tumor grows, urinary tract symptoms (such as frequent urination, urgency, difficulty urinating, interrupted urine flow, frequent waking up at night, etc.), bone pain (bone metastasis), hematuria or hematospermia may occur.

[0007] Prostate cancer is usually screened and diagnosed through the following methods: Prostate-specific antigen (PSA) test: PSA is a protein secreted by the prostate. PSA levels are usually elevated in patients with prostate cancer. PSA tests are used for early screening, but they are not completely reliable because certain benign diseases can also cause PSA to be elevated. Digital rectal examination (DRE): The doctor examines the prostate by feeling the rectum to feel for lumps or abnormal enlargement. Prostate ultrasound: It can be used to evaluate the size and shape of the prostate and is sometimes used to guide a biopsy. Prostate biopsy: A biopsy is usually performed under the guidance of a rectal ultrasound to collect prostate tissue for pathological examination to confirm whether it is cancer. Imaging tests: Such as CT, MRI, or bone scans are used to evaluate the spread of cancer, especially if the cancer is suspected to have spread.

[0008] Currently, the treatment of prostate cancer depends on factors such as the stage of cancer, the patient's age, health status, and whether there is distant metastasis. Common treatments include surgery, radiotherapy, hormone therapy, chemotherapy, targeted therapy, and immunotherapy. Although existing treatments can effectively treat some patients, for some patients with advanced or drug-resistant prostate cancer, the treatment effect is limited and the side effects are large. Therefore, finding new treatment approaches and drugs has become an urgent clinical need. Summary of the invention

[0009] In view of this, the present invention provides an application of a madder extract in the preparation of a drug for treating tumors. The madder extract of the present invention has higher targeting and specificity, and has low side effects and high tolerance, and has a good effect in the treatment of urinary system tumors. The extraction method of the present invention ensures the retention of effective ingredients, improves the quality of the drug, and provides a new potential treatment option for the treatment of urinary system tumors and other tumors.

[0010] The invention provides application of madder extract in preparing medicine for treating tumors.

[0011] Preferably, the tumor is a urinary system tumor.

[0012] Preferably, the Rubia cordifolia extract further comprises pharmaceutically acceptable excipients.

[0013] Preferably, the madder extract is prepared from madder slices by water extraction and alcohol precipitation.

[0014] In the prior art, drugs for treating urinary system tumors mainly rely on treatment methods such as chemical drugs and radiotherapy, which often have significant side effects. The Rubia cordifolia extract provided by the present invention has anti-tumor potential, especially for urinary system tumors, and its pharmacological mechanism includes inhibiting tumor cell proliferation, inducing tumor cell apoptosis, inhibiting tumor metastasis, etc., providing a new treatment option for the treatment of urinary system tumors.

[0015] As a natural medicine, the madder extract of the present invention is generally better than traditional chemical drugs in terms of safety and tolerance, has lower toxic and side effects, has good tolerance, and can reduce treatment discomfort and side effects of patients.

[0016] The invention adopts a water extraction and alcohol precipitation method to prepare a madder extract, which can effectively extract active ingredients in the madder, retain its anti-tumor active ingredients, and the method is relatively simple and environmentally friendly. Compared with the traditional extraction method, the water extraction and alcohol precipitation method can better retain the active ingredients in the madder, thereby ensuring the effectiveness of the drug and the quality stability and consistency of the madder extract.

[0017] The active ingredients in the Rubia cordifolia extract of the present invention can better exert their anti-tumor effects, especially for urinary system tumors, and may inhibit tumor cell proliferation, induce apoptosis, inhibit metastasis and other effects through multi-target effects. This makes the Rubia cordifolia extract a potential anti-tumor drug, especially suitable for tumor types that are insensitive to existing treatment methods or have unsatisfactory treatment effects.

[0018] The present invention is mainly aimed at urinary system tumors, but the anti-tumor effect of madder extract has a wide potential and may not be limited to the treatment of urinary system tumors. Its multi-mechanism anti-tumor effect may be extended to other types of tumor treatment fields, providing more options for cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The present invention will be further described below in conjunction with the accompanying drawings.

[0020] Figure 1 The inhibitory effects of different concentrations of Rubia cordifolia extract on the proliferation of PC3, DU145, C4-2B, and 22RV1 cell lines;

[0021] Figure 2 The results of observation under objective microscope before and after the effect of 160μg / mL Rubia cordifolia extract on PC3, DU145, C4-2B, and 22RV1 cell lines;

[0022] Figure 3 This is the result of the scratch test;

[0023] Figure 4 The inhibitory effect of different concentrations of Rubia cordifolia extract on the proliferation of OSRC2 cell line;

[0024] Figure 5 The inhibitory effect of different concentrations of Rubia cordifolia extract on the proliferation of 786-O cell line;

[0025] Figure 6 The inhibitory effect of different concentrations of Rubia cordifolia extract on the proliferation of T24 cell line;

[0026] Figure 7 The inhibitory effect of different concentrations of Rubia cordifolia extract on the proliferation of U3 cell line;

[0027] Figure 8 The figure shows the inhibitory effect of different concentrations of Rubia cordifolia extract on the proliferation of 5637 cell line. DETAILED DESCRIPTION

[0028] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] The invention provides application of madder extract in preparing medicine for treating tumors.

[0030] Rubia cordifolia L. is a plant of the genus Rubia in the Rubiaceae family. It is also known as blood madder and blood-seeing sorrow. Rubia cordifolia has medicinal value. According to the "Chinese Materia Medica Dictionary", its functions and indications are: "Cooling blood to stop bleeding, promoting blood circulation and removing blood stasis. It is mainly used to treat blood heat, hemoptysis, hematemesis, hematuria, bloody stool, metrorrhagia, amenorrhea, abdominal pain caused by blood stasis after childbirth, traumatic injuries, rheumatic pain, jaundice, sores, carbuncle, and hemorrhoids." In addition, Rubia cordifolia has certain economic value. Rubia cordifolia root can be used as a red dye to dye animal or plant fibers. Pliny's "Natural History" also records that Rubia cordifolia can be used to dye wool and leather.

[0031] In the field of traditional Chinese medicine, the root and rhizome of Rubia cordifolia are used as medicine. It tastes bitter, is cold in nature, and belongs to the liver meridian. It has the effects of cooling blood, activating blood circulation, removing blood stasis, and regulating menstruation. It is used for vomiting blood, epistaxis, metrorrhagia, traumatic bleeding, amenorrhea, joint pain, and swelling and pain due to falls.

[0032] Unlike traditional Chinese medicine, which uses madder to cool blood and stop bleeding, promote blood circulation and remove blood stasis, and remove blood stasis and promote menstruation, the present invention explores the potential of madder extract in tumor treatment through modern science and technology and pharmacological research. The new use of madder of the present invention not only broadens the boundaries of its traditional medicinal use, but also provides new ideas for the treatment of urinary system tumors and other types of tumors. Through the research of madder active ingredients by modern science and technology, its potential in tumor treatment is found, which provides a new drug selection for clinical use. Through further clinical trials and verification, madder extract is expected to become a safe and effective anti-tumor drug, especially in anti-urinary system tumors, with broad application prospects.

[0033] The tumor of the present invention is preferably a urinary system tumor, and the urinary system tumor is preferably prostate cancer.

[0034] The Rubia cordifolia extract of the present invention also includes pharmaceutically acceptable excipients, such as various organic or inorganic carrier substances commonly used as preparation materials, which are used as excipients, lubricants, adhesives, disintegrants, thickeners in solid preparations; and solvents, dispersants, solubilizers, suspending agents, tonicity agents, buffers, analgesics, etc. in liquid preparations. In addition, additives such as preservatives, antioxidants, colorants, sweeteners, etc. can be used according to conventional methods as needed.

[0035] In some specific embodiments of the present invention, the excipient is at least one of microcrystalline cellulose, lactose, pregelatinized starch, white sugar, D-mannitol, starch, and light anhydrous silicic acid; the lubricant is at least one of magnesium stearate, calcium stearate, talc, polyethylene glycol, silicon dioxide, sodium stearyl fumarate, micropowder silica gel, and hydrogenated vegetable oil; the binder is at least one of hydroxypropyl methylcellulose, povidone, pregelatinized starch, methylcellulose, sodium carboxymethyl cellulose, gum arabic, gelatin, polyvinyl alcohol, microcrystalline cellulose, and polyethylene glycol; the disintegrant is cross-linked sodium carboxymethyl cellulose, cross-linked povidone, low-substituted hydroxypropyl The invention relates to a novel nanostructured carbon foam, wherein the nanostructured carbon foam is selected from the group consisting of cellulose, corn starch, microcrystalline cellulose, sodium bicarbonate, sodium carboxymethyl starch, hydroxypropyl methylcellulose, carboxymethyl cellulose and citric acid; the thickener is selected from the group consisting of hydroxypropyl methylcellulose, polyvinyl alcohol, carbomer, xanthan gum, guar gum, sodium alginate, gum arabic, gelatin, methylcellulose, sodium carboxymethyl cellulose and polyethylene glycol; the solvent is selected from the group consisting of water for injection, alcohol, propylene glycol, polyethylene glycol, dimethyl sulfoxide, isopropyl myristate, vegetable oil, mineral oil and ethyl acetate; the dispersant is selected from the group consisting of polyvinyl pyrrolidone, hydroxypropyl methylcellulose, polyethylene glycol, Tween The solubilizing agent is at least one of Tween 80, sodium lauryl sulfate, lecithin, polyoxyethylene fatty alcohol ether, Tween 20, Tween 80, Span 20, Span 80, polyoxyethylene sorbitan monooleate, magnesium aluminum silicate, and silicon dioxide; the solubilizing agent is at least one of Tween 80, Tween20, Span 20, Span 80, Brij 35, polyoxyethylene sorbitan monooleate, lecithin, ethanol, glycerol, polyvinyl pyrrolidone, hydroxypropyl methylcellulose, sodium lauryl sulfate, Poloxamer 188, D-mannitol, benzyl benzoate, triethanolamine, sodium carbonate, and sodium citrate; the suspending agent is hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, xanthan gum, sodium alginate, magnesium aluminum silicate, polyvinyl alcohol, povidone, polyethylene glycol, gelatin, gum arabic, Tween 80. At least one of dodecyl sulfuric acid and silicon dioxide; the surfactant is at least one of polyvinyl alcohol, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose; the tonicity agent may be at least one of sodium chloride, glucose, mannitol, sorbitol, sucrose, sodium lactate, phosphate buffer, potassium chloride, calcium chloride, magnesium chloride and glycerol.

[0036] The madder extract of the present invention is prepared from madder pieces by water extraction and alcohol precipitation, which specifically comprises the following steps:

[0037] The decoction pieces of Rubia cordifolia are decocted and filtered, anhydrous ethanol is poured into the filtrate, the mixture is allowed to stand for precipitation and then filtered, the ethanol is removed and the mixture is dried to obtain the Rubia cordifolia extract.

[0038] In some preferred embodiments of the present invention, the Rubia cordifolia slices need to be soaked before decocting, and the soaking time is not less than 30 minutes.

[0039] In some specific embodiments of the present invention, the preparation method of the Rubia cordifolia extract is as follows:

[0040] Add 8 to 10 times the volume of water to the Rubia cordifolia slices, soak at room temperature for 30 to 60 minutes, condense and reflux and decoct for 60 to 90 minutes, filter, repeat the condense and reflux and decoction for 30 to 45 minutes, combine the filtrate, add 5 to 8 times the volume of anhydrous ethanol after cooling, let stand at 0°C to 4°C for 36 to 48 hours, filter, remove the solvent and freeze-dry to obtain the Rubia cordifolia extract.

[0041] The madder extract prepared in the specific example of the present invention is stored at -80°C.

[0042] In order to further illustrate the present invention, the following examples are provided for detailed description. The raw materials used in the following examples of the present invention are all commercially available.

[0043] Unless otherwise specified, all experiments were repeated three times. SPSS 21.0 was used for analysis of variance (ANOVA) and Duncan's multiple comparison analysis. The results were expressed as mean ± standard deviation, and P < 0.05 indicated a significant difference.

[0044] Embodiment 1 A method for preparing a Rubia cordifolia extract, comprising the following steps:

[0045] Add 10 times the volume of water to the Rubia cordifolia slices and soak them at room temperature for 45 minutes, condense and reflux and decoct for 60 minutes, filter, repeat the condense and reflux and decoction for 45 minutes, combine the filtrate, add 8 times the volume of anhydrous ethanol after cooling, let stand at 4°C for 48 hours, filter, remove the solvent and freeze-dry to obtain the Rubia cordifolia extract.

[0046] Test Example 1 Inhibitory effect of Rubia cordifolia extract on the proliferation of prostate cancer cells

[0047] PC3 is a human prostate adenocarcinoma cell line that exhibits high proliferation and invasiveness in vitro. p53 gene mutations are often detected in PC3 cell lines, and they usually lack androgen receptor (AR) expression, making them insensitive to androgens.

[0048] DU145 is a human prostate adenocarcinoma cell line that exhibits high proliferation and invasiveness in vitro. p53 gene mutations are also often detected in the DU145 cell line, which also lacks androgen receptor (AR) expression and is insensitive to androgens.

[0049] The C4-2B cell line was isolated from LNCaP cells after long-term androgen deprivation therapy. The androgen receptor (AR) expression in the C4-2B cell line was high, but the sensitivity to androgen was low.

[0050] The 22RV1 cell line is isolated from LNCaP cells after long-term androgen deprivation therapy and has strong androgen resistance. The androgen receptor (AR) expression in the 22RV1 cell line is high, but the sensitivity to androgen is low, and it can secrete PSA (prostate-specific antigen).

[0051] PC3, DU145, C4-2B, and 22RV1 prostate cancer cell line models were selected, and the cells were treated with different concentrations (10, 20, 40, 80, and 160 μg / mL) of Rubia cordifolia extract (prepared in Example 1) for 48 h. The CCK-8 results are as follows: Figure 1 shown.

[0052] Depend on Figure 1 Combined with the microscopic observation results ( Figure 2 ) showed that the Rubia cordifolia extract of the present invention can significantly inhibit the proliferation of prostate cancer PC3, DU145, C4-2B, and 22RV1 cells.

[0053] Test Example 2 Effect of Rubia cordifolia extract on PC3, C4-2B and 22RV1 cell metastasis

[0054] The effect of the Rubia cordifolia extract (prepared in Example 1) on prostate cancer metastasis was tested by a scratch test. The results are as follows: Figure 3 shown.

[0055] Depend on Figure 3 It can be seen that the Rubia cordifolia extract of the present invention can significantly inhibit the metastasis of different prostate cancer cells.

[0056] Test Example 3 Inhibitory effect of Rubia cordifolia extract on RM-1 cell proliferation

[0057] Cell culture: Mouse prostate cancer cell line RM-1 is an adherent cell, cultured in IMDM medium containing 10% calf serum, placed in a 5% CO2, 37°C constant temperature sealed incubator, subcultured once every 48-72 hours, and digested with 0.25% trypsin (containing 0.02% EDTA). Cells in the logarithmic growth phase were taken and divided into an experimental group and a blank control group.

[0058] The specific method of cell passaging is as follows: select cells in the logarithmic growth phase and pass them when the cells reach 80% fusion. Gently shake the culture bottle several times to suspend the debris attached to the cell surface, pour it out together with the culture medium, and wash it twice with PBS to remove the residual serum. Add 1mL of 0.25% trypsin digestion solution and observe it under a microscope. When the cells gradually become round, add culture medium containing calf serum to stop digestion, blow the cells to suspend them, transfer them to a centrifuge tube, centrifuge at 800rpm for 5min, discard the supernatant, wash once with PBS, add 5mL of IMDM culture medium containing 10% calf serum, transfer them to a culture bottle, and culture them in an incubator. The passaging ratio is 1:3.

[0059] The cells were digested with 0.25% trypsin and centrifuged. 1 mL of -20°C precooled freezing solution was added. After being pipetted evenly, the cells were transferred into cryotubes. After sealing and marking, the cells were placed at 4°C for 40 min, -20°C for 30-60 min, and -80°C overnight. Cells used within one month can be stored at -80°C. For long-term storage, the cells can be transferred into liquid nitrogen after 24 h.

[0060] When in use, take out the cryopreservation tube containing cells from liquid nitrogen, immediately put it into 37℃ warm water and shake it quickly. When the cryopreservation liquid is dissolved by more than 50%, transfer it into 5mL IMDM culture medium preheated at 37℃ in the clean bench, and ensure that it is rewarmed within 5 minutes. Use a pipette to blow evenly, centrifuge at 800rpm for 5 minutes, discard the supernatant, wash once with PBS, add 5mL IMDM culture medium, transfer it to a culture bottle, and culture it in an incubator.

[0061] MTT assay:

[0062] RM-1 cells in the logarithmic growth phase were obtained and digested with trypsin. The cell concentration was adjusted to 5 × 10 4 / mL, 100μL cell suspension was added to each well of a sterile 96-well culture plate. After 24h, the cells adhered to the wall and the culture medium was discarded. 180μL of culture medium was added to each well. The experimental group added 20μL of culture medium containing different doses of madder extract, and the control group added 20μL of culture medium. A total of 8 groups were divided, namely the control group, madder extract (prepared in Example 1) 10μg / mL group, 20μg / mL group, 40μg / mL group, 80μg / mL group, 160μg / mL group, each group was set with 4 replicates, and was divided into 12h, 24h, and 48h groups according to the action time of madder extract. 4h before the end of the drug action, 20μL of MTT (5mg / mL) was added to each well. After 4h of MTT action, the culture medium was discarded, and 150μL of DMSO was added to each well. The cells were shaken for 10min, and the absorbance value (OD570) of each well was detected by an ELISA instrument to calculate the cell growth inhibition rate.

[0063] Cell growth inhibition rate (%) = (1-experimental group / control group) × 100%

[0064] Table 1 Proliferation inhibition rate of RM-1 cells after 12h treatment with Rubia cordifolia extract

[0065]

[0066] * indicates P < 0.05 compared with the control group, ** indicates P < 0.01 compared with the control group.

[0067] The inhibitory effects of different concentrations of Rubia cordifolia extract on RM-1 cells after 12h, 24h and 48h are shown in Table 1. As shown in Table 1, the inhibitory effect of the Rubia cordifolia extract of the present invention on RM-1 cell proliferation is obviously time-dose dependent.

[0068] Test Example 4 Effect of Rubia cordifolia extract on RM-1 cell cycle

[0069] The subcultured RM-1 cells were digested with 0.25% trypsin and counted. The cells were inoculated into culture flasks at a rate of 1×10 cells per flask. 6 , add madder extract after 24 hours, the dosage is 160μg / mL, the final volume of the culture medium is 4mL, after 24 hours, collect the cells, centrifuge, blow, and wash twice with cold PBS. Suspend the above cells with 1mL70% cold ethanol and fix at 4℃ for 24 hours. After digestion with RNase, add 1.5mLPI (50mg / L) to stain DNA. After mixing, perform single parameter analysis on the flow cytometer, and calculate the proportion of cells in each stage by cumulative curve segmentation method.

[0070] The results show that there is no G1 phase arrest in the blank control group. After 24h of the action of the madder extract on RM-1 cells, the percentage of cells in the G1 phase is higher than that in the control group, and the percentage of cells in the G1 phase is (37.42 ± 0.85)%. After 48h of the action of the madder extract on RM-1 cells, the cell cycle is mainly blocked in the G1 phase, and the percentage of cells in the G1 phase is (49.86 ± 0.64)%. It can be seen that the madder extract of the present invention can significantly inhibit the G1 phase process of RM-1 cells, and as the treatment time is extended, the blocking effect of the G1 phase gradually increases. After 48 hours, the inhibitory effect of the madder extract on G1 phase cells is more significant, suggesting that it may affect the proliferation of RM-1 cells through G1 phase arrest.

[0071] Test Example 5 Inhibitory Effect of Rubia cordifolia Extract on Proliferation of Renal Cancer Cells

[0072] OSRC-2 is a human renal clear cell carcinoma cell line that exhibits high proliferation and invasiveness in vitro. VHL gene mutations are often detected in OSRC-2 cell lines, which are common genetic changes in clear cell renal carcinoma.

[0073] 786-O is a human renal clear cell carcinoma cell line that also exhibits high proliferation and invasiveness in vitro. The 786-O cell line lacks VHL gene function, which is also one of its important molecular characteristics. In addition, p53 gene mutations are often detected in the 786-O cell line.

[0074] OSRC2 and 786-O renal cancer cell line models were selected, and cells were treated with different concentrations (10, 20, 40, 80, 160 μg / mL) of Rubia cordifolia extract (prepared in Example 1) for 48 h. The CCK-8 results were as follows: Figure 4 and Figure 5 shown.

[0075] Depend on Figure 4 and Figure 5 It can be seen that the Rubia cordifolia extract of the present invention can significantly inhibit the proliferation of renal cancer cell lines OSRC2 and 786-O.

[0076] Test Example 6 Inhibitory effect of Rubia cordifolia extract on proliferation of bladder cancer cells

[0077] T24 is a human bladder transitional cell carcinoma, in which p53 gene mutations are frequently detected, a common genetic alteration in bladder cancer. In addition, the T24 cell line also expresses high levels of epidermal growth factor receptor (EGFR).

[0078] U3 is another human bladder transitional cell carcinoma cell line that exhibits certain proliferation and invasiveness in vitro. Some gene mutations associated with bladder cancer, such as p53 gene mutations, are often detected in the U3 cell line.

[0079] 5637 is a human bladder transitional cell carcinoma cell line. p53 gene mutations are often detected in 5637 cell line, which usually expresses high levels of epidermal growth factor receptor (EGFR) and chromosomal aneuploidy is often observed in the cell line, which is related to its highly malignant phenotype.

[0080] T24, U3, and 5637 bladder cancer cell line models were selected, and the cells were treated with different concentrations (10, 20, 40, 80, and 160 μg / mL) of Rubia cordifolia extract (prepared in Example 1) for 48 h. The CCK-8 results were as follows: Figure 6-8 shown.

[0081] Depend on Figure 6-8 It can be seen that the Rubia cordifolia extract of the present invention can significantly inhibit the proliferation of bladder cancer cell lines T24, U3, and 5637.

[0082] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Application of Rubia cordifolia extract in the preparation of drugs for treating tumors.

2. The use according to claim 1, characterized in that: The tumor is a urinary system tumor.

3. The use according to claim 1, characterized in that: The madder extract is prepared from madder pieces through water extraction and alcohol precipitation.

4. The use according to claim 3, characterized in that: The water extraction and alcohol precipitation method comprises the following steps: The decoction pieces of Rubia cordifolia are decocted and filtered, anhydrous ethanol is poured into the filtrate, the mixture is allowed to stand for precipitation and then filtered, the ethanol is removed and the mixture is dried to obtain the Rubia cordifolia extract.

5. The use according to claim 4, characterized in that: Soak the Rubia cordifolia slices before decocting.

6. The use according to claim 5, characterized in that: During the soaking process, the volume ratio of Rubia cordifolia slices to water is 1:(8~10), the soaking temperature is room temperature, and the soaking time is 30~60 min.

7. The use according to claim 4, characterized in that: The decoction is condensation reflux decoction, and the decoction is performed twice, the first decoction time is 60-90 min, and the second decoction time is 30-45 min.

8. The use according to claim 4, characterized in that: The volume ratio of the filtrate to anhydrous ethanol is 1:(5-8), the standing temperature is 0°C-4°C, and the standing time is 36-48 h.

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