Use of sodium polystyrene sulfonate in preparation of drug for treating hepatocellular carcinoma

Abstract

Disclosed in the present invention is use of sodium polystyrene sulfonate (PSS) in the preparation of a drug for treating hepatocellular carcinoma. According to the use, PSS is used as an active ingredient for treating hepatocellular carcinoma. Also provided is a drug for treating hepatocellular carcinoma, the active ingredient of which comprises sodium polystyrene sulfonate. PSS can effectively inhibit the invasion and migration of liver cancer cells in vitro, as well as reduce the relative content of potassium ions and depolarize the cell membrane in HCC cell lines, thereby significantly inhibiting the progression of liver cancer. Moreover, PSS has the advantages of high safety, few side effects, wide sources, and easy synthesis. Therefore, PSS can effectively treat hepatocellular carcinoma, and can effectively prevent metastasis of hepatocellular carcinoma, thereby preventing hepatocellular carcinoma from progressing from early to intermediate and advanced stages. PSS has great application prospects in the preparation of a drug for treating hepatocellular carcinoma.

Description

Application of sodium polystyrene sulfonate in the preparation of drugs for the treatment of hepatocellular carcinoma Technical Field

[0001] This invention relates to the field of hepatocellular carcinoma treatment drugs, and more particularly to the use of sodium polystyrene sulfonate in the preparation of hepatocellular carcinoma treatment drugs. Background Technology

[0002] Primary liver cancer refers to malignant tumors that occur in hepatocytes or intrahepatic bile duct cells. Statistics show that approximately 85% of primary liver cancers are hepatocellular carcinoma (HCC). Due to its insidious onset, over 70% of HCC patients are diagnosed at an intermediate or advanced stage, resulting in a poor prognosis. Therefore, preventing the occurrence of HCC or preventing its progression from early to intermediate or advanced stages is crucial. Consequently, finding effective drugs to prevent HCC metastasis and reduce the risk for high-risk groups is of great significance.

[0003] Sodium polystyrene sulfonate (PSS) is a water-soluble polymer with strong dispersing and emulsifying properties. It can be used as a water treatment agent, emulsifier, and cation exchange resin. In the pharmaceutical field, PSS has the effects of lowering blood lipids, blood pressure, and blood sugar. In addition, it has certain applications in the preparation of anticancer drug delivery carriers. However, its own effects on cancer have not been reported. Summary of the Invention

[0004] To address the aforementioned problems, this invention provides the application of sodium polystyrene sulfonate (PSS) in the preparation of hepatocellular carcinoma therapeutic drugs, wherein PSS is used as an active ingredient for the treatment of hepatocellular carcinoma, rather than as a carrier of the active ingredient.

[0005] Furthermore, the application uses PSS as an effective component to inhibit the invasion or migration of liver cancer cells.

[0006] Furthermore, the application uses PSS as an effective ingredient for inhibiting liver cancer metastasis.

[0007] Preferably, the molecular weight of the PSS is 10,000 to 100,000, and more preferably 50,000 to 80,000.

[0008] The present invention also provides a drug for treating hepatocellular carcinoma, wherein the active ingredient of the drug includes sodium polystyrene sulfonate.

[0009] Furthermore, the drug also includes pharmaceutically acceptable carriers or excipients.

[0010] Furthermore, the drug is used to inhibit the invasion or migration of liver cancer cells.

[0011] Furthermore, the drug is used to inhibit liver cancer metastasis.

[0012] Furthermore, the drug is used to prevent hepatocellular carcinoma from progressing from its early stages to its middle and late stages.

[0013] The inventors discovered that PSS can effectively inhibit the invasion and migration of liver cancer cells in vitro, and significantly inhibit liver cancer development by reducing the relative potassium ion content and depolarizing the cell membrane in HCC cell lines. Furthermore, PSS has the advantages of high safety, few side effects, wide availability, and easy synthesis. Therefore, PSS can effectively treat hepatocellular carcinoma and prevent its metastasis, thereby preventing the progression of hepatocellular carcinoma from early to advanced stages. It shows great promise in the preparation of drugs for treating hepatocellular carcinoma.

[0014] The following will further explain the concept, specific structure, and technical effects of the present invention in conjunction with the accompanying drawings, so as to fully understand the purpose, features, and effects of the present invention. Attached Figure Description

[0015] Figure 1 shows the experimental results of the effect of PSS on cell migration in Example 1; the left side shows the cell migration results of HepG2 cells after the addition of PSS; the right side shows the cell migration results of PLC cells after the addition of PSS; the figure was drawn by GraphPadprism 8.0 (*p<0.05, **p<0.001, ***p<0.0001, ****p<0.00001);

[0016] Figure 2 shows the experimental results of the effect of PSS on cell invasion in Example 2; the left side shows the cell invasion results of HepG2 cells after the addition of PSS; the right side shows the cell invasion results of PLC cells after the addition of PSS; the figure was drawn by GraphPadprism 8.0 (*p<0.05, **p<0.001, ***p<0.0001, ****p<0.00001);

[0017] Figure 3 is a statistical graph of the experimental results of the change of relative potassium ion content in cells by PSS in Example 3; In the figure, A is the relative potassium ion content in HepG2 cells after adding PSS and detecting it with a potassium ion fluorescent probe; B is the relative potassium ion content in PLC cells after adding PSS and detecting it with a potassium ion fluorescent probe; The figure was drawn by GraphPadprism 8.0 (*p<0.05, **p<0.001, ***p<0.0001, ****p<0.00001);

[0018] Figure 4 is a statistical graph of the experimental results of the change in cell polarity caused by PSS in Example 4; A in the figure is the change in cell membrane polarity of HepG2 cells after adding PSS and detecting it with DiBAC4(3); B is the change in cell membrane polarity of PLC cells after adding PSS and detecting it with DiBAC4(3); the figure was drawn by GraphPadprism8.0 (*p<0.05, **p<0.001, ***p<0.0001, ****p<0.00001). Detailed Implementation

[0019] Sodium polystyrene sulfonate (PSS) used in this embodiment was purchased from Sigma, catalog number 243051, with a molecular weight of approximately 70,000.

[0020] Example 1: Effect of PSS on Cancer Cell Migration

[0021] The migration ability of human hepatocellular carcinoma (HCC) cells (PLC and HepG2 cell lines) was detected using the Transwell Assay. Cells were treated with or without PSS, using 8 μm pore size chambers (greiner bio-one, 662638) and 24-well transwell plates (greiner bio-one, 662160).

[0022] Once the cells have reached the logarithmic growth phase, digest the cells, wash them once with ice-cold 1×PBS, resuspend them in FBS-free high-glucose medium, and count and seed them. Add 5×10⁶ cells to the upper chamber. 4 Cells were cultured in 200 μL FBS-free high-glucose medium, with 500 μL of complete medium (10% FBS + 1% PS + high-glucose DMEM) added to the lower chamber. The PSS group received a final PSS concentration of 10 nM, while the other group received no PSS. Each group was tested in triplicate. After 48 hours of incubation at 37°C, cells that had not migrated through the upper chamber were wiped off with a cotton swab. The lower surface of the chamber was fixed with 4% paraformaldehyde at room temperature for 15 minutes, followed by staining with 0.1% crystal violet at room temperature for 15 minutes. Residual staining was washed off the cells, and the cells were inverted and air-dried for microscopic examination and photographic analysis. The results are shown in Figure 1. The left side shows the cell migration assay results for HepG2 cells after PSS addition, and the right side shows the cell migration assay results for PLC cells after PSS addition. The figures show that the number of migrating cells in the 10 nM PSS group was significantly lower than that in the group without PSS. These results indicate that the migration ability of both cancer cell types is reduced after the addition of PSS.

[0023] Example 2: Effect of PSS on cell invasion

[0024] The invasive ability of human hepatocellular carcinoma (HCC) cells (HepG2 and PLC cell lines) was detected using the Transwell Assay. Cells were treated with PSS (10 nM) or without PSS. 8 μm pore size chambers (greiner bio-one, 662638) and 24-well transwell plates (greiner bio-one, 662160) were used.

[0025] Thaw the matrix gel (Corning, 356231) on ice. Dilute the matrix gel with ice-cold FBS-free high-glucose DMEM medium (gel:FBS-free medium = 1:11). Add 100 μL of the diluted matrix gel to the upper chamber and incubate at 37°C for 2 hours. Digest the cells, wash them once with ice-cold 1×PBS, resuspend them in FBS-free high-glucose DMEM medium, and count and seed them. Add 5×10⁶ cells to the upper chamber. 4 Cells were cultured in 200 μL FBS-free high-glucose medium. 500 μL of DMEM medium containing 10% FBS, 1% PS, and high-glucose was added to the lower chamber. The PSS group received a final PSS concentration of 10 nM, while the other group received no PSS. Each group was configured with three replicates. After 48 hours of incubation at 37°C, cells that had not penetrated the membrane in the upper chamber were wiped off with a cotton swab. The lower surface of the chamber was fixed with 4% paraformaldehyde at room temperature for 15 minutes, followed by crystal violet staining at room temperature for 15 minutes. Residual staining solution was washed off the cell surfaces, and the cells were inverted and air-dried for microscopic examination and photographic analysis. The results are shown in Figure 2. The left side shows the cell invasion assay results for HepG2 cells after PSS addition; the right side shows the cell invasion assay results for PLC cells after PSS addition. The figures show that the number of invasive cells in the 10 nM PSS group was significantly lower than that in the group without PSS. These results indicate that the invasive ability of both cancer cells was reduced after the addition of PSS.

[0026] Example 3: Relationship between PSS and intracellular potassium ions in cancer cells

[0027] Human hepatocellular carcinoma (HCC) cells, specifically the HepG2 and PLC cell lines, were used. Cells were treated with or without PSS, and the relative intracellular potassium ion content was compared. The relative intracellular potassium ion content was detected using a potassium ion fluorescent probe (IPG-4AM, ION Biosciences, 0321F).

[0028] Specific detection steps: When the two types of HCC cells are in the logarithmic growth phase, after digestion, the cells are resuspended in 10% FBS + 1% PS high-glucose DMEM. Count 10,000 cells (100 μL) per well and seed them into 96-well plates. Add 100 μL of reagent to each well to divide the cells into three groups with PSS final concentrations of 10 nM, 5 nM, and 0 nM. Set up 3 replicates for each group. Incubate at 37℃ for 1 hour. Detect the absorbance of the samples at 350 nm and 380 nm using a microplate reader (model).

[0029] The results are shown in Figure 3. In Figure A, the relative intracellular potassium content of HepG2 cells after the addition of PSS was detected using a potassium ion fluorescent probe; in Figure B, the relative intracellular potassium content of PLC cells after the addition of PSS was detected using a potassium ion fluorescent probe. The figures show that the relative intracellular potassium content in both cancer cells was significantly reduced after the addition of PSS. This reduction in potassium ions can promote apoptosis and thus inhibit cancer cell growth.

[0030] Example 4: Relationship between PSS and cancer cell membrane polarity

[0031] Human liver cancer cells (HCC) were selected: HepG2 and PLC cell lines. Cells were treated with or without PSS, and changes in cell membrane polarity were compared. Among them, the changes in cell membrane polarity were detected using the potentiometric probe bis(1,3-dibarbituric acid)-trimethyloxenolol (DiBAC4(3), Thermo Fisher Scientific, B438).

[0032] Specific detection steps: When the two types of HCC cells are in the logarithmic growth phase, after digestion, the cells are resuspended in 10% FBS + 1% PS high glucose DMEM. Count 10,000 cells (100 μL) per well and seed them into 96-well plates. Add 100 μL of reagent to each well to divide the cells into three groups with final PSS concentrations of 10 nM, 5 nM, and 0 nM. Set up three replicates for each group. Incubate at 37°C for 1 hour. Detect the absorbance of the samples at 540 nm using a microplate reader (model).

[0033] The results are shown in Figure 4. In Figure A, the cell membrane polarity changes of HepG2 cells after the addition of PSS were detected using DiBAC4(3); and in PLC cells, the cell membrane polarity changes of PLC cells after the addition of PSS were detected using DiBAC4(3). As can be seen from the figure, the addition of PSS to the two cancer cells disrupted the intracellular ion balance, caused hyperpolarization of the cells, remodeled the extracellular matrix, inhibited EMT transformation, and inhibited the growth of cancer cells.

[0034] The preferred embodiments of the present invention have been described in detail above. It should be understood that those skilled in the art can make numerous modifications and variations based on the concept of the present invention without creative effort. Therefore, all technical solutions that can be obtained by those skilled in the art based on the concept of the present invention through logical analysis, reasoning, or limited experimentation on the basis of existing technology should be within the scope of protection defined by the claims.

Claims

1. Use of sodium polystyrene sulfonate in the preparation of a drug for treating hepatocellular carcinoma, characterized in that, The application uses PSS as an effective component for hepatocellular carcinoma treatment.

2. The use of sodium polystyrene sulfonate according to claim 1 for the preparation of a medicament for the treatment of hepatocellular carcinoma, characterized in that, The application uses PSS as an effective component for inhibiting invasion or migration of liver cancer cells.

3. The use of sodium polystyrene sulfonate according to claim 1 for the preparation of a medicament for the treatment of hepatocellular carcinoma, characterized in that, The application uses PSS as an effective component for inhibiting metastasis of liver cancer.

4. A medicament for treating hepatocellular carcinoma, characterized by comprising a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof: ###0000014### (I) The effective component includes sodium polystyrene sulfonate.

5. The medicament for treating hepatocellular carcinoma according to Claim 4, wherein A pharmaceutically acceptable carrier or excipient is also included.

6. The medicament for treating hepatocellular carcinoma according to Claim 4, wherein The medicine is used for inhibiting invasion or migration of liver cancer cells.

7. The medicament for treating hepatocellular carcinoma according to Claim 4, wherein The medicine is used for inhibiting metastasis of liver cancer.

8. The medicament for treating hepatocellular carcinoma according to Claim 4, wherein The medicine is used for preventing hepatocellular carcinoma from developing from early stage to middle-late stage.

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