Application of oldenlandia diffusa in preparation of medicine for treating liver cancer

Abstract

The invention discloses application of oldenlandia diffusa in preparation of a medicine for treating liver cancer. On the first aspect, the invention provides application of oldenlandia diffusa in preparation of drugs for promoting apoptosis and / or active oxygen level of liver cancer cells. It is found in the experiment process that oldenlandia diffusa can be independently used as a Chinese herbal medicine preparation for treating liver cancer and can effectively induce apoptosis and oxidative stress of liver cancer cells.

Description

Technical Field

[0001] The present application relates to the technical field of liver cancer, and in particular to the application of Hedyotis diffusa in the preparation of medicines for treating liver cancer. Background Art

[0002] Hepatocellular carcinoma (HCC) accounts for about 90% of primary liver cancer cases. The 5-year overall survival rate of HCC patients in China is only 12%. Most HCC patients are already in the advanced stage when diagnosed, so curative resection is not feasible. In addition, the current clinical chemotherapy drugs are ineffective, and the overall high recurrence rate has not been improved. Therefore, there is an urgent need to develop new treatments to optimize the clinical treatment of HCC. Traditional Chinese medicine is considered to be useful for the treatment of HCC due to its multi-target and coordinated intervention characteristics. Many studies have shown that phytochemical and molecular biological treatments based on Chinese herbal compounds have prospects in the preparation of anti-HCC products. Some in vitro and in vivo experimental results have shown that traditional Chinese medicine can inhibit the growth of HCC cells by inducing cell death and immune regulation, inhibiting metastasis, alleviating inflammatory responses, and improving antiviral activity. In recent years, several retrospective cohort studies have also shown that combined treatment with traditional Chinese medicine can improve the survival rate of all HCC patients, including patients with unresectable lesions after hepatic artery chemoembolization and radiofrequency ablation of liver cancer.

[0003] Hedyotis diffusa Willd (HDW) is a Chinese herbal medicine recorded in the Chinese Pharmacopoeia (2015 edition), which has the functions of clearing away heat and detoxifying, promoting blood circulation and removing blood stasis, and promoting diuresis and relieving stranguria. Studies have shown that HDW can induce cell apoptosis and enhance the immune function of cells, indicating that it has anti-cancer activity. At the same time, more and more reports have demonstrated the anti-tumor effect of HDW in a series of cancers such as colorectal cancer, cervical cancer, glioblastoma, lung cancer, breast cancer and leukemia. In the treatment of liver cancer, HDW has also been widely used as an adjuvant therapeutic agent for liver cancer. It is reported that HDW induces apoptosis of HCC cells through the AKT / mTOR and JNK / Nur77 signaling pathways. At the same time, HDW enhances the efficacy of sorafenib and 5-fluorouracil in the treatment of HCC and improves side effects. However, due to its multi-target characteristics, other molecular mechanisms of HDW in the treatment of HCC are not yet fully understood. Therefore, further exploration of the relevant molecular mechanisms of HDW and finding molecular targets for liver cancer will provide more options for the clinical treatment of patients. Summary of the invention

[0004] The present application aims to solve at least one of the technical problems existing in the prior art. To this end, the present application proposes an application of Hedyotis diffusa in the preparation of a drug for treating liver cancer.

[0005] In a first aspect of the present application, a use of Hedyotis diffusa in preparing a drug that promotes apoptosis and / or reactive oxygen levels in liver cancer cells is provided.

[0006] According to the use of the first aspect of the present application, the following beneficial effects are achieved:

[0007] During the experiment, it was found that Houttuynia cordata can be used alone as a Chinese herbal preparation for the treatment of liver cancer, and can effectively induce apoptosis and oxidative stress in liver cancer cells.

[0008] In some embodiments of the present application, the liver cancer is hepatocellular carcinoma.

[0009] In some embodiments of the present application, the liver cancer cells are at least one of Huh7 and MHCC97.

[0010] In some embodiments of the present application, the liver cancer cell MHCC97 includes at least one of MHCC97H and MHCC97L. The two cell lines obtained by screening the parental strain MHCC97 are named MHCC97H and MHCC97L according to the metastatic potential. Therefore, the three differ only in metastatic potential and other properties are relatively consistent.

[0011] In some embodiments of the present application, Hedyotis diffusa is used in the preparation of a drug in the form of Hedyotis diffusa preparation.

[0012] In some embodiments of the present application, the preparation of Hedyotis diffusa includes: liquid preparations, such as low molecular weight solutions (solutions, aromatic waters, syrups, spirits, tinctures, glycerin), polymer solutions, sols, suspensions, mixtures, lotions, liniments, coatings, ear drops, ear washes, nasal drops, nasal washes, enemas; solid preparations, such as powders, granules, capsules, tablets, pills, and pills. In some embodiments, the preparation of Hedyotis diffusa is a sterile preparation, such as an injection, an infusion, a sterile powder for injection, and any of other sterile preparations (such as an implanted drug delivery system, a wound preparation, a sponge), etc. In some embodiments, the preparation of Hedyotis diffusa is a preparation for skin and mucous membranes, such as a suppository, an ointment and a cream, a gel, a film, an aerosol, a spray, a powder aerosol, etc. In some embodiments, the Hedyotis diffusa preparation is a nano preparation, such as any one of nanoparticles, solid lipid nanoparticles, nanocrystals, polymer micelles, nanolipid discs, etc. In some embodiments, the Hedyotis diffusa preparation is a sustained-release and controlled-release preparation, such as any one of sustained-release and controlled-release matrix tablets, sustained-release and controlled-release capsules, osmotic pump tablets, sustained-release and controlled-release implants, delayed-release preparations (enteric-coated capsules and tablets, colon-localized preparations, pulse preparations), etc.

[0013] In some embodiments of the present application, the Hedyotis diffusa preparation is Hedyotis diffusa injection.

[0014] In some embodiments of the present application, the raw material of the Hedyotis diffusa injection includes Hedyotis diffusa.

[0015] In some embodiments of the present application, the raw materials of Hedyotis diffusa injection also include auxiliary materials.

[0016] In some embodiments of the present application, the excipient includes at least one of a pharmaceutically acceptable carrier, a diluent, a stabilizer, a filler, a wetting agent, a colorant, a solvent, a co-solvent, a film former, a spontaneity promoter, an emulsifier, a dispersant, a preservative, an antifreeze agent, a thickener, and an adjuvant.

[0017] In some embodiments of the present application, the auxiliary material includes at least one of polysorbate and sodium bisulfite.

[0018] In a second aspect of the present application, a use of Hedyotis diffusa in preparing a medicament for inhibiting the activation of hepatic stellate cells is provided.

[0019] In some embodiments of the present application, the activation of hepatic stellate cells is the activation of hepatic stellate cells stimulated by liver cancer cells.

[0020] In some embodiments of the present application, the liver cancer is hepatocellular carcinoma.

[0021] In some embodiments of the present application, the liver cancer cells are at least one of Huh7 and MHCC97.

[0022] In some embodiments of the present application, the liver cancer cell MHCC97 includes at least one of MHCC97H and MHCC97L.

[0023] In some embodiments of the present application, the stimulation of liver cancer cells is the co-culture of liver cancer cells and hepatic stellate cells to be stimulated.

[0024] In some embodiments of the present application, the stimulation of liver cancer cells includes hepatic stellate cells in the tumor microenvironment.

[0025] In some embodiments of the present application, Hedyotis diffusa is used in the preparation of a drug in the form of Hedyotis diffusa preparation.

[0026] In some embodiments of the present application, the preparation of Hedyotis diffusa includes: liquid preparations, such as low molecular weight solutions (solutions, aromatic waters, syrups, spirits, tinctures, glycerin), polymer solutions, sols, suspensions, mixtures, lotions, liniments, coatings, ear drops, ear washes, nasal drops, nasal washes, enemas; solid preparations, such as powders, granules, capsules, tablets, pills, and pills. In some embodiments, the preparation of Hedyotis diffusa is a sterile preparation, such as an injection, an infusion, a sterile powder for injection, and any of other sterile preparations (such as an implanted drug delivery system, a wound preparation, a sponge), etc. In some embodiments, the preparation of Hedyotis diffusa is a preparation for skin and mucous membranes, such as a suppository, an ointment and a cream, a gel, a film, an aerosol, a spray, a powder aerosol, etc. In some embodiments, the Hedyotis diffusa preparation is a nano preparation, such as any one of nanoparticles, solid lipid nanoparticles, nanocrystals, polymer micelles, nanolipid discs, etc. In some embodiments, the Hedyotis diffusa preparation is a sustained-release and controlled-release preparation, such as any one of sustained-release and controlled-release matrix tablets, sustained-release and controlled-release capsules, osmotic pump tablets, sustained-release and controlled-release implants, delayed-release preparations (enteric-coated capsules and tablets, colon-localized preparations, pulse preparations), etc.

[0027] In some embodiments of the present application, the Hedyotis diffusa preparation is Hedyotis diffusa injection.

[0028] In some embodiments of the present application, the raw material of the Hedyotis diffusa injection includes Hedyotis diffusa.

[0029] In some embodiments of the present application, the raw materials of Hedyotis diffusa injection also include auxiliary materials.

[0030] In some embodiments of the present application, the excipient includes at least one of a pharmaceutically acceptable carrier, a diluent, a stabilizer, a filler, a wetting agent, a colorant, a solvent, a co-solvent, a film former, a spontaneity promoter, an emulsifier, a dispersant, a preservative, an antifreeze agent, a thickener, and an adjuvant.

[0031] In some embodiments of the present application, the auxiliary material includes at least one of polysorbate and sodium bisulfite.

[0032] The third aspect of the present application provides a use of a substance that inhibits the level of ADH4 in the preparation of a drug for treating liver cancer.

[0033] In some embodiments of the present application, the substance that inhibits the level of ADH4 includes at least one of the following:

[0034] A substance that specifically inhibits the knockdown or knockout of the gene encoding ADH4; a substance that specifically inhibits the transcription level of ADH4; a substance that specifically inhibits the protein expression level of ADH4; a substance that specifically inhibits the protein activity of ADH4.

[0035] In some embodiments of the present application, the substance includes Hedyotis diffusa.

[0036] In some embodiments of the present application, the substance includes an Oldenlandia diffusa preparation.

[0037] In some embodiments of the present application, the preparation of Hedyotis diffusa includes: liquid preparations, such as low molecular weight solutions (solutions, aromatic waters, syrups, spirits, tinctures, glycerin), polymer solutions, sols, suspensions, mixtures, lotions, liniments, coatings, ear drops, ear washes, nasal drops, nasal washes, enemas; solid preparations, such as powders, granules, capsules, tablets, pills, and pills. In some embodiments, the preparation of Hedyotis diffusa is a sterile preparation, such as an injection, an infusion, a sterile powder for injection, and any of other sterile preparations (such as an implanted drug delivery system, a wound preparation, a sponge), etc. In some embodiments, the preparation of Hedyotis diffusa is a preparation for skin and mucous membranes, such as a suppository, an ointment and a cream, a gel, a film, an aerosol, a spray, a powder aerosol, etc. In some embodiments, the Hedyotis diffusa preparation is a nano preparation, such as any one of nanoparticles, solid lipid nanoparticles, nanocrystals, polymer micelles, nanolipid discs, etc. In some embodiments, the Hedyotis diffusa preparation is a sustained-release and controlled-release preparation, such as any one of sustained-release and controlled-release matrix tablets, sustained-release and controlled-release capsules, osmotic pump tablets, sustained-release and controlled-release implants, delayed-release preparations (enteric-coated capsules and tablets, colon-localized preparations, pulse preparations), etc.

[0038] In some embodiments of the present application, the Hedyotis diffusa preparation is Hedyotis diffusa injection.

[0039] In some embodiments of the present application, the raw material of the Hedyotis diffusa injection includes Hedyotis diffusa.

[0040] In some embodiments of the present application, the raw materials of Hedyotis diffusa injection also include auxiliary materials.

[0041] In some embodiments of the present application, the excipient includes at least one of a pharmaceutically acceptable carrier, a diluent, a stabilizer, a filler, a wetting agent, a colorant, a solvent, a co-solvent, a film former, a spontaneity promoter, an emulsifier, a dispersant, a preservative, an antifreeze agent, a thickener, and an adjuvant.

[0042] In some embodiments of the present application, the auxiliary material includes at least one of polysorbate and sodium bisulfite.

[0043] In some embodiments of the present application, the substance includes a nucleic acid molecule for RNA interference.

[0044] In some embodiments of the present application, the nucleic acid molecule for RNA interference includes at least one of siRNA, shRNA, miRNA, and dsRNA for RNA interference of ADH4.

[0045] In some embodiments of the present application, the substance further includes a carrier that carries the nucleic acid molecule.

[0046] In some embodiments of the present application, the vector includes any one of a plasmid, a lentivirus, an adenovirus, and an adeno-associated virus.

[0047] In some embodiments of the present application, the nucleotide sequence of the sense strand of the siRNA is as shown in SEQ ID NO.1; and / or, the nucleotide sequence of the antisense strand of the siRNA is as shown in SEQ ID NO.2.

[0048] In the examples of this application, bioinformatics screening and in vitro experiments were used to explain the molecular mechanism of HDW in inhibiting the occurrence and development of liver cancer, and the target genes of HDW were screened. The experimental results revealed the role of HDW alone as a Chinese herbal preparation for the treatment of liver cancer, as well as the molecular mechanism of HDW in the activation process of hepatic stellate cells and the screening of target genes, and based on this, gene sets with HCC prognostic value and HDW target genes were mined, and more drugs with corresponding therapeutic effects for hepatocellular carcinoma can also be developed using the target gene sites.

[0049] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become apparent from the description below, or will be learned through the practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 This is the sensitivity test result of different types of liver cancer cells to HDW in one embodiment of the present invention.

[0051] Figure 2 This is the apoptosis detection result of liver cancer cells after HDW treatment in one embodiment of the present invention.

[0052] Figure 3 This is the result of detecting active oxygen in liver cancer cells after HDW treatment in one embodiment of the present invention.

[0053] Figure 4 This is the sensitivity test result of activated LX-2 cells to HDW in one embodiment of the present invention.

[0054] Figure 5 This is the test result of migration and invasion assay of hepatic stellate cells in one embodiment of the present invention.

[0055] Figure 6This is the result of screening and pathway enrichment analysis of HDW target gene set in liver cancer in one embodiment of the present invention. Wherein, A is a Venn diagram of HDW target and HCC differential genes, and B is the result of pathway enrichment analysis of HDW-TG.

[0056] Figure 7 This is a survival analysis and expression analysis of the HDW target gene set in liver cancer in one embodiment of the present invention, wherein A is the ROC curve of different genes, and B is the ADH4 expression results in HCC tissues with different degrees of differentiation.

[0057] Figure 8 This is the activity detection result of ADH4 silenced cells in one embodiment of the present invention.

[0058] Fig. 9 This is the detection result of the migration and invasion experiment of ADH4 silenced cells in one embodiment of the present invention. DETAILED DESCRIPTION

[0059] The following will clearly and completely describe the concept of the present application and the technical effects produced in combination with the embodiments, so as to fully understand the purpose, features and effects of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of them. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present application.

[0060] The embodiments of the present application are described in detail below. The described embodiments are exemplary and are only used to explain the present application, and should not be understood as limiting the present application.

[0061] In the description of this application, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, "above", "below", "within", etc. are understood to include the number itself, and "about" means within the range of ±20%, 10%, 8%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, 0.1%, etc. of the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0062] In the description of the present application, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0063] The present application is described below in conjunction with specific embodiments.

[0064] Some of the materials, reagents and instruments involved are described as follows:

[0065] Human hepatoma cell lines Huh7 and MHCC97H;

[0066] Human normal hepatic stellate cell line LX-2;

[0067] Hedyotis diffusa injection (National Medicine Standard No. Z34020595)

[0068] DMEM medium (Thermo Fisher Scientific, USA)

[0069] Dichlorofluorescein diacetate (Biyuntian, S0033, China)

[0070] siRNA (Hanheng Biotechnology Shanghai Co., Ltd.)

[0071] Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific, USA)

[0072] CCK-8 kit (Biyuntian, C0038, China)

[0073] Annexin V-FITC apoptosis detection kit (Biyuntian, C1062S)

[0074] Transwell chamber (8 μm pore size, Corning, USA)

[0075] Reconstituted Matrigel (#356234, Corning, USA)

[0076] Fluorescence microscope (Leica, DMi8, Germany)

[0077] Multiskan GO microplate reader (Thermo Fisher Scientific, USA)

[0078] Example 1: HDW inhibits the activity of liver cancer cells

[0079] Huh7 cells and MHCC97H cells were inoculated in 96-well plates, respectively. After 24 hours of inoculation, the culture medium was replaced with 100 μL of fresh culture medium (DMEM containing 10% FBS) containing 0wt% HDW (NC group), 5wt% HDW, 10wt% HDW and 20wt% HDW. After incubation for 48 hours, the cell activity was detected using the CCK-8 kit according to the instructions.

[0080] The results are as follows Figure 1 As shown, HDW significantly inhibited the activity of both types of liver cancer cells within the dosage range of 5wt% to 20wt%.

[0081] Example 2: HDW induces apoptosis of liver cancer cells

[0082] Huh7 cells and MHCC97H cells were inoculated in 6-well plates and treated with medium (DMEM containing 10% FBS) containing 0wt% HDW (control group, NC) and 10wt% HDW at 37°C and 5% CO2 for 48 hours. After washing the cells with PBS, the cells were stained with Annexin V-FITC and propidium iodide (PI) using the Annexin V-FITC apoptosis detection kit according to the instructions. Finally, fluorescence photos were taken using a fluorescence microscope (Leica, DMi8, Germany).

[0083] The results are as follows Figure 2 As shown, in the above two types of liver cancer cells, compared with the control group, the number of apoptotic cells in the 10wt% HDW group increased greatly, indicating that the addition of HDW can induce apoptosis of liver cancer cells.

[0084] Example 3: HDW induces oxidative stress in liver cancer cells

[0085] The liver cancer cells were treated with HDW according to the method of Example 2, then washed with PBS according to the instructions, and incubated with 10 μM dichlorofluorescein diacetate at 37° C. for 30 min. Then, the cells were washed twice with PBS, and the absorbance was detected using Multiskan GO (excitation wavelength of 488 nm, emission wavelength of 525 nm).

[0086] The results are as follows Figure 3 As shown, the levels of reactive oxygen species in both liver cancer cells treated with HDW were significantly increased compared with those in control cells.

[0087] Example 4: HDW inhibits the activation of hepatic stellate cells

[0088] In order to study whether the HDW targeted gene is related to the activation of normal human hepatic stellate cells (HSC), the sensitivity of LX-2 cells to HDW was detected by cell viability experiment according to the method of Example 1.

[0089] The results are as follows Figure 4 As shown in the figure, compared with normal LX-2 cells, LX-2 cells activated by TGF-β1 or liver cancer cell culture supernatant are more sensitive to HDW. Among them, TGF-β1 can regulate cell proliferation, growth, differentiation and cell movement, activate inflammatory factors in LX-2 cells, and have a profibrotic effect on LX-2 cells; while the liver cancer cell culture supernatant contains a large number of cellular inflammatory factors, which can promote the proliferation, migration and invasion of LX-2 cells, and also has a profibrotic effect on LX-2 cells.

[0090] The invasion and migration abilities of cancer cells were evaluated by measuring the number of cells passing through the Transwell chamber.

[0091] Among them, the steps of the cell invasion experiment are as follows: liver cancer cells are inoculated in a Transwell chamber with a recombinant matrigel-coated membrane on the upper layer, and the number of invaded cancer cells is counted after incubation for 48 hours;

[0092] The steps of the cell migration experiment are as follows: liver cancer cells are inoculated in a Transwell chamber without a recombinant matrix gel coating membrane, and the number of migrated cancer cells is counted after incubation for 24 hours.

[0093] The migrated or invaded cells on the bottom of the chamber were fixed with 70% ethanol, stained with hematoxylin and eosin, and counted in 5 random fields using a microscope.

[0094] In the co-culture experiment, liver cancer cells were first plated in a 24-well plate or pretreated with 5% HDW for 24 hours. Hepatic stellate cells LX-2 were then added to the Transwell chamber, and then liver cancer cells were inoculated into the Transwells. According to the above invasion and migration ability experiments, the number of migrated cancer cells was counted after 24 hours of incubation, and the number of invaded cancer cells was counted after 48 hours of incubation.

[0095] The results are as follows Figure 5 As shown, 5% HDW did not attenuate the migration or invasion ability of LX-2 cells by Transwell assay. In co-culture Transwell assay, MHCC97H or Huh7 cells increased the migration and invasion ability of LX-2 cells, while 5% HDW inhibited the enhanced migration and invasion ability of LX-2 cells under co-culture.

[0096] Example 5

[0097] After predicting the target genes of each component of HDW in the BATMAN-TCM database (http: / / bionet.ncpsb.org.cn / batman-tcm / ), the HCC-related differentially expressed genes were retrieved in the GEPIA2 database (http: / / gepia2.cancer-pku.cn / #index), and a Venn diagram was drawn for the two groups of genes.

[0098] The GSVA algorithm was used in the GSCA database (http: / / bioinfo.life.hust.edu.cn / GSCA / # / ) to perform enrichment analysis of cancer-related pathways on the intersecting gene sets.

[0099] The gene expression data of HCC tissues and healthy tissues were obtained from the TCGA database (https: / / portal.gdc.cancer.gov / ), and bioinformatics analyses such as gene expression analysis and survival analysis were performed on the intersecting gene sets using SRplot (http: / / www.bioinformatics.com.cn / srplot).

[0100] The results showed that after searching HDW in the BATMAN-TCM database and merging duplicates, the total number of predicted HDW targets was 168; at the same time, the GEPIA2 database was used to search for 2206 differentially expressed genes related to hepatocellular carcinoma. By drawing a Venn diagram of HDW targets and HCC differentially expressed genes, 24 intersecting genes were screened out. The results are as follows Figure 6 A.

[0101] Pathway enrichment analysis was performed on these 24 gene sets (HDW-TGS) using gene set cancer analysis. The results are as follows Figure 6 B, indicating that HDW-TGS is highly correlated with estrogen receptor signaling pathway, Ras-MAPK pathway, and RTK signaling pathway, indicating that HDW may target genes related to cell proliferation and differentiation in liver cancer.

[0102] Using HCC sample data from the TCGA database (from TCGA's Liver Hepatocellular Carcinoma, a total of 379 samples), an overall survival analysis of all genes in HDW-TGS was performed, and it was found that 8 of them were independently correlated with the overall survival of HCC patients, including ADH1B, ADH4, ATP1A1, APOE, CYP2E1, ESR1, GJA5, and IGF1. ROC curves were drawn based on the expression levels of these genes in HCC tissues and healthy tissues. The results are shown in Figure 2. Figure 7As shown in A, the bottom area of ​​the curve is greater than 0.7, indicating that these genes have potential prognostic value for HCC. In addition, the expression of ADH4 in HCC tissues with different degrees of differentiation was compared. Figure 7 As shown in B, there are significant differences in HCC tissues with different degrees of differentiation, which indicates that ADH4 is related to the occurrence and development of HCC.

[0103] Example 6: ADH4 Function Verification

[0104] To further investigate the role of ADH4 as a HDW target gene in the progression of liver cancer, we constructed an ADH4-knockdown liver cancer cell line by transfecting small interfering RNA (siRNA). The specific process is as follows: siRNA transfection was completed using Lipofectamine 3000 transfection reagent, and after antibiotic screening, an ADH4-knockdown cell line (siADH4) and a control cell line (NC) were obtained.

[0105] The sequence of siRNA is as follows:

[0106] Sense strand: 5'-CCAGCAGCCCGAGGUCUUCTT-3' (SEQ ID NO. 1), wherein TT is a protruding overhang end;

[0107] Anti-sense strand (anti-sense): 5'-GAAGACCUCGGGCUGCUGGTT-3' (SEQ ID NO. 2), wherein TT is a protruding overhang end.

[0108] Referring to Examples 1 and 4, cell viability, migration and invasion experiments were performed on HDW-treated Huh7 cells and ADH4 knockdown Huh7 cells (siADH4). Figure 8 and Fig. 9 As shown, the results showed that after silencing ADH4 using the above siRNA, the viability, migration and invasion abilities of liver cancer cells were significantly reduced.

[0109] The present application is described in detail above in conjunction with the embodiments, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present application. In addition, the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

Claims

1. Use of Hedyotis diffusa in preparing a drug that promotes apoptosis and / or reactive oxygen levels in liver cancer cells.

2. The use according to claim 1, characterized in that The liver cancer is hepatocellular carcinoma.

3. The use according to claim 1, characterized in that The liver cancer cells are at least one of Huh7 and MHCC97.

4. Use of Hedyotis diffusa in preparing a drug for inhibiting the activation of hepatic stellate cells.

5. The use according to claim 4, characterized in that The activation of hepatic stellate cells is the activation of hepatic stellate cells stimulated by liver cancer cells.

6. The use according to claim 5, characterized in that The liver cancer cells are at least one of Huh7 and MHCC97.

7. Use of a substance that inhibits the level of ADH4 in the preparation of a drug for treating liver cancer.

8. The use according to claim 7, characterized in that: The substance includes oldenlandia diffusa.

9. The use according to claim 7, characterized in that: The substances include siRNA of ADH4.

10. The use according to claim 9, characterized in that: The nucleotide sequence of the sense strand of the siRNA is shown as SEQ ID NO.1, and / or the nucleotide sequence of the antisense strand of the siRNA is shown as SEQ ID NO.2.

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