Application of mPGES-2 as target spot in preparation of medicine for preventing and / or treating liver cancer

Abstract

The invention discloses an application of mPGES-2 as a target spot in preparation of a medicine for preventing and / or treating liver cancer. The invention proposes that mPGES-2 is a drug target for preventing and / or treating liver cancer for the first time. Experiments show that mPGES-2 knock-down significantly down-regulates expression of a cell proliferation antigen Ki67 and DNA replication marker protein PCNA, inhibits the S-phase process of cells, and further inhibits proliferation slowing caused by DNA replication of liver cancer cells; overexpression of mPGES-2 can obviously promote expression up-regulation of Ki67 and PCNA, promote the S-phase process of cells, enhance the DNA replication efficiency and finally cause obvious increase of cell proliferation; the mPGES-2 can regulate the proliferation of the liver cancer cells, so that the mPGES-2 can be used as a drug target for preventing and / or treating the liver cancer.

Description

Technical Field

[0001] The present invention relates to the application of mPGES-2 (microsomal prostaglandin E synthase-2) as a target in the preparation of drugs for preventing and / or treating liver cancer, belonging to the field of biomedical technology. Background Art

[0002] Hepatocellular carcinoma (HCC) is the main form of primary liver cancer, currently accounting for approximately 85% to 90% of all liver cancer patients. The occurrence of HCC is closely related to a variety of risk factors, including but not limited to chronic viral hepatitis, cirrhosis, alcoholic liver disease, non-alcoholic fatty liver disease, and exposure to aflatoxin, etc. The biological process of HCC is extremely complex, involving the interaction of multiple factors, which together lead to the imbalance between the inactivation of tumor suppressor genes and the activation of oncogenes, ultimately resulting in the abnormal activation of molecular signaling pathways, thus triggering the occurrence of liver cancer. At present, the treatment methods for liver cancer have covered various methods such as surgical operation, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. However, due to the lack of obvious clinical symptoms in the early stage of liver tumors and the insufficient understanding of their genetic and molecular characteristics, the effective treatment of HCC still faces huge challenges. Therefore, finding new drug targets and developing drugs for treating liver cancer have become important topics in the field of medical research, and the progress in this field will bring new hope and vitality to tens of thousands of patients. Summary of the Invention

[0003] The main purpose of the present invention is to provide an application of mPGES-2 as a target in the preparation of drugs for preventing and / or treating liver cancer, so as to overcome the deficiencies in the prior art.

[0004] To achieve the foregoing invention purpose, the technical solutions adopted by the present invention include:

[0005] The embodiments of the present invention provide an application of mPGES-2 as a target in the preparation of drugs for preventing and / or treating liver cancer.

[0006] The embodiments of the present invention also provide an application of an inhibitor of mPGES-2 or its coding gene in the preparation of drugs for preventing and / or treating liver cancer.

[0007] The embodiments of the present invention also provide a pharmaceutical composition for preventing and / or treating liver cancer, which comprises: an inhibitor of mPGES-2 or its coding gene, and a pharmaceutically acceptable carrier.

[0008] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention reports for the first time the application of mPGES-2 as a drug target in the prevention and / or treatment of liver cancer. Experiments show that knockdown of mPGES-2 significantly downregulates the expression of cell proliferation antigen Ki67 and DNA replication marker protein PCNA, inhibits the cell S-phase progression, and further inhibits DNA replication in liver cancer cells, resulting in slowed proliferation. Overexpression of mPGES-2 can significantly promote the upregulation of Ki67 and PCNA expression, promote the cell S-phase progression, enhance the DNA replication efficiency, and ultimately cause a significant increase in cell proliferation. These results indicate that mPGES-2 can regulate the proliferation of liver cancer cells, suggesting that mPGES-2 can be used as a drug target for the prevention and / or treatment of liver cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments recorded in the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0010] Figure 1a It is a diagram showing the expression results of mPGES-2 in mouse liver cancer cells Hepa1-6 in the control group (negative control, NC) and the mPGES-2 knockdown (KD) group in a typical embodiment of the present invention;

[0011] Figure 1b It is a statistical chart of the crystal violet experiment of mouse liver cancer cells Hepal-6 in the control group and the mPGES-2 KD group in a typical embodiment of the present invention;

[0012] Figure 1c It is a statistical chart of cell proliferation detected by the CCK-8 method for mouse liver cancer cells Hepal-6 in the control group and the mPGES-2 KD group in a typical embodiment of the present invention;

[0013] Figure 2a It is a diagram showing the expression results of mPGES-2 in mouse liver cancer cells Hepa1-6 in the control group and the mPGES-2 overexpression (OE) group in a typical embodiment of the present invention;

[0014] Figure 2b It is a statistical chart of the crystal violet experiment of mouse liver cancer cells Hepa1-6 in the control group and the mPGES-2 OE group in a typical embodiment of the present invention;

[0015] Figure 2c It is a statistical chart of cell proliferation detected by the CCK-8 method for mouse liver cancer cells Hepa1-6 in the control group and the mPGES-2 OE group in a typical embodiment of the present invention;

[0016] Figure 3a It is a result graph of detecting the DNA replication efficiency of mouse liver cancer cells Hepa1-6 in the control group and the mPGES-2KD group by using the Click it EdU experiment in a typical embodiment of the present invention;

[0017] Figure 3b It is a result graph of detecting the DNA replication efficiency of mouse liver cancer cells Hepa1-6 in the control group and the mPGES-2OE group by using the Click it EdU experiment in a typical embodiment of the present invention;

[0018] Figure 4a It is a result graph of the expression of proliferation-related protein Ki67 and DNA replication marker protein PCNA in mouse liver cancer cells Hepal-6 in the control group and the mPGES-2KD group in a typical embodiment of the present invention;

[0019] Figure 4b It is a result graph of the expression of proliferation-related protein Ki67 and DNA replication marker protein PCNA in mouse liver cancer cells Hepal-6 in the control group and the mPGES-2OE group in a typical embodiment of the present invention;

[0020] Figure 5a It is a result graph of detecting the cell cycle of mouse liver cancer cells Hepal-6 after mPGES-2 knockdown by using flow cytometry in a typical embodiment of the present invention;

[0021] Figure 5b It is a result graph of detecting the cell cycle of mouse liver cancer cells Hepa1-6 after mPGES-2 overexpression by using flow cytometry in a typical embodiment of the present invention;

[0022] Figure 6a It is a result graph of detecting the nuclear entry of mPGES-2 in mouse liver cancer cells Hepa1-6 by using nuclear-cytoplasmic separation in a typical embodiment of the present invention;

[0023] Figure 6b It is a result graph of detecting the chromatin binding of mPGES-2 in mouse liver cancer cells Hepa1-6 by using chromatin extraction in a typical embodiment of the present invention. Detailed implementation manners

[0024] In view of the defects of the prior art, the inventors of this case have proposed the technical solution of the present invention through long-term research and a large number of practices. To facilitate the understanding of this application, the following will describe this application in more detail. However, it should be understood that the present invention can be implemented in many different forms and is not limited to the embodiments or examples described herein. On the contrary, the purpose of providing these embodiments or examples is to make the understanding of the disclosure content of the present invention more thorough and comprehensive.

[0025] Specifically, as an aspect of the technical solution of the present invention, it relates to the use of mPGES-2 as a target in the preparation of a drug for preventing and / or treating liver cancer.

[0026] In some preferred embodiments, the knockdown of the target can at least inhibit the proliferation of liver cancer cells in liver cancer mice.

[0027] In some preferred embodiments, the knockdown of the target can at least inhibit the DNA replication of liver cancer cells in liver cancer mice.

[0028] In some preferred embodiments, the knockdown of the target can at least down-regulate the expression of Ki67 and PCNA in liver cancer cells of liver cancer mice.

[0029] In some preferred embodiments, the overexpression of the target can at least promote the proliferation of liver cancer cells in liver cancer mice.

[0030] In some preferred embodiments, the overexpression of the target can at least promote the DNA replication of liver cancer cells in liver cancer mice.

[0031] In some preferred embodiments, the overexpression of the target can at least up-regulate the expression of Ki67 and PCNA in liver cancer cells of liver cancer mice.

[0032] In some preferred embodiments, the knockdown of the target can at least reduce the proportion of S-phase liver cancer cells in liver cancer mice.

[0033] In some preferred embodiments, the overexpression of the target can at least increase the proportion of S-phase liver cancer cells in liver cancer mice.

[0034] In some preferred embodiments, the mPGES-2 can at least enter the nucleus of liver cancer cells and bind to chromatin.

[0035] Furthermore, the knockdown of mPGES-2 can at least inhibit the proliferation of liver cancer cells.

[0036] Furthermore, the overexpression of mPGES-2 can at least promote the proliferation of liver cancer cells.

[0037] Furthermore, the knockdown of mPGES-2 can at least inhibit the DNA replication efficiency of liver cancer cells.

[0038] Furthermore, the overexpression of mPGES-2 can at least improve the DNA replication efficiency of liver cancer cells.

[0039] Furthermore, the inhibition of mPGES-2 can at least inhibit the expression of PCNA and Ki67 in liver cancer cells.

[0040] Furthermore, overexpression of mPGES-2 can at least increase the expression of PCNA and Ki67 in liver cancer cells.

[0041] Furthermore, inhibition of mPGES-2 can at least reduce the proportion of liver cancer cells in the S phase.

[0042] Furthermore, overexpression of mPGES-2 can at least increase the proportion of liver cancer cells in the S phase.

[0043] Furthermore, mPGES-2 can at least enter the nucleus and bind to chromatin.

[0044] As another aspect of the technical solution of the present invention, it relates to the use of an inhibitor of mPGES-2 or its coding gene in the preparation of a drug for preventing and / or treating liver cancer.

[0045] In some preferred embodiments, the inhibitor is selected from interfering molecules that specifically interfere with the expression of the coding gene of mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its coding gene.

[0046] As another aspect of the technical solution of the present invention, it also relates to a pharmaceutical composition for preventing and / or treating liver cancer, which comprises: an inhibitor of mPGES-2 or its coding gene, and a pharmaceutically acceptable carrier.

[0047] In some preferred embodiments, the inhibitor is selected from interfering molecules that specifically interfere with the expression of the coding gene of mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its coding gene.

[0048] The present invention is further illustrated by the following examples: According to the following examples, the present invention can be better understood. However, those skilled in the art can easily understand that the specific material ratios, process conditions and their results described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0049] Unless otherwise specified, the various raw materials, reaction equipment, test equipment and test methods used in the following examples are well-known in the art.

[0050] Example 1: Experiment on detecting the effect of mPGES-2 knockdown or overexpression on the proliferation of liver cancer cells

[0051] a. Cell line construction

[0052] Mouse hepatocarcinoma cell line Hepa 1-6 control (NC), mPGES-2 knockdown (KD), and mPGES-2 overexpression (OE) cell line models were constructed by infecting mPGES-2 NC, mPGES-2 KD, and mPGES-2 OE lentiviruses into mouse hepatocarcinoma cells Hepa 1-6.

[0053] b. Detection of virus infection in mouse hepatocarcinoma cells Hepa 1-6 by Western Blot

[0054] (1) RIPA lysis buffer (brand: Beyotime) (add 10 μL of 100 mM PMSF per milliliter of lysis buffer before use) was added to mPGES-2 NC and mPGES-2 KD cells. After lysis on ice for 30 min, the supernatant was collected by centrifugation at 12,000 rpm for 15 min.

[0055] (2) Quantification was performed using a BCA protein concentration detection kit (brand: Thermofisher). The supernatant was mixed with 5× loading buffer at a ratio of 1:4, vortexed, and heated at 100 °C in a metal bath for 10 min.

[0056] (3) Total proteins were separated by 10% SDS-PAGE gel under the separation conditions: 150 V, 90 min, and then wet transferred to a nitrocellulose membrane under the wet transfer conditions: 80 V, 90 min. The membrane was blocked with PBST solution containing 5% bovine serum albumin V (brand: Solarbio) for 1 h, incubated with rabbit anti-mPGES-2 antibody overnight, washed with TBST 3 times, 5 min each time; incubated with secondary antibody at room temperature for 1 h; washed with TBST 3 times, 5 min each time, and the signal was detected using an ECL chemiluminescence solution (brand: Fluorescence). The expression results of mPGES-2 are as Figure 1a shown.

[0057] c. Detection of the effect of mPGES-2 knockdown on the proliferation of mouse hepatocarcinoma cells Hepa 1-6 by crystal violet assay

[0058] (1) Cell suspensions of Hepa 1-6 NC / KD / OE in the logarithmic growth phase were seeded into 6-well culture plates (1×10 3 cells / well), and 2 mL of high-glucose DMEM culture medium was added to each well. The plates were cultured in an incubator at 37 °C and 5% CO2 for 7 days.

[0059] (2) The old culture medium was discarded, and each well was washed once with 1× PBS and then fixed with 2 mL of methanol for 15 min.

[0060] (3) Methanol was discarded, and 2 mL of 1% crystal violet solution was added to each well for staining for 1 h.

[0061] (4) Discard the 1% crystal violet solution, rinse with running water, and take a photo for record.

[0062] The results of the crystal violet experiment on the proliferation of hepatoma cells by mPGES-2 are as Figure 1b 、 Figure 2b shown.

[0063] d. Detect the effect of mPGES-2 knockdown on the proliferation of mouse hepatoma cells Hepa 1-6 by CCK8 assay

[0064] (1) Inoculate the logarithmic growth phase Hepa 1-6 NC / KD / OE cell suspension into a 96-well culture plate (2×10 3 cells / well), and add 100 μL of high-glucose DMEM culture medium (containing 10% fetal bovine serum and 1% penicillin and streptomycin mixture) to each well. Incubate in a 37°C, 5% CO2 incubator.

[0065] (2) After 4, 24, 48, 72, 96, and 120 h respectively, add 10 μL of CCK-8 (brand: Yeason) detection solution to each well, and incubate at 37°C in the dark for 60 min. Set up blank control wells (containing the same culture medium and CCK-8), with 5 replicates in each group, and repeat the experiment three times.

[0066] (3) Use an enzyme-linked immunosorbent assay (ELISA) reader to detect the OD value of each well, with a detection wavelength of 450 nm.

[0067] The results of the CCK8 assay on the proliferation of hepatoma cells by mPGES-2 are as Figure 1c 、 Figure 2c shown.

[0068] Example 2: Experiment to detect the effect of mPGES-2 knockdown or overexpression on cell DNA replication

[0069] After EdU with azide binds to DNA, it reacts with alkyne with fluorescence to label the replicated DNA.

[0070] (1) Plate mPGES-2 NC, mPGES-2 KD, and mPGES-2 OE cells, add cell slides, with a cell density of 3×10 5 / well / 2 mL (6-well plate), and culture in normal complete medium for 24 h;

[0071] (2) Culture the negative control with 3 mM HU (Hydroxyurea, brand: Sigma) for 1 h;

[0072] (3) EdU staining:

[0073] a. Add EdU (brand: Invitrogen) at a final concentration of 40 μM and culture for 20 min;

[0074] b. Discard the old culture medium and wash once with 2 mL of 1×PBS;

[0075] c. Perform pre-extraction (this step is carried out on ice and all reagents are pre-cooled):

[0076] Wash each well once with 2 mL of 1×CSK and remove;

[0077] Add CSK-0.5% Triton and incubate on ice for 5 min, then remove;

[0078] Add 1×CSK, 2 mL per well, leave for a short time and then aspirate clean;

[0079] Add 1×PBS, 2 mL per well, leave for a short time and then aspirate clean;

[0080] d. Fixation: Add 1 mL of 4% PAF paraformaldehyde and fix at room temperature for 7 min;

[0081] Remove 4% PAF paraformaldehyde, add 1×PBS and wash 2 times, standing for 2 min each time;

[0082] e. Prepare Click-it reaction cocktails (brand: Invitrogen), 50 μL of the mixture is required for each sample, add the mixture, protect from light at room temperature for 30 min, and wash once with PBS;

[0083] (4) Stain PCNA:

[0084] Add pre-cooled methanol and place at -20 °C for 15 min;

[0085] Wash twice with PBS, 2 min each time;

[0086] Block with PBST containing 5% BSA at room temperature for 1 h;

[0087] Primary antibody diluted with PBST containing 5% BSA: PCNA (brand: Santa cruz), diluted 1:50, incubate at room temperature for 1 h;

[0088] Wash 3 times with PBST, 2 min each time;

[0089] Secondary antibody (50 μL) diluted with PBST containing 5% BSA, diluted 1:200, incubate at room temperature for 30 min, PBST

[0090] Wash 2 times, 2 min each time;

[0091] Stain with DAPI (brand: Sigma) (1:500), incubate at room temperature for 5 min;

[0092] Wash three times with PBS, 2 min each time;

[0093] Mount the slides and observe under a microscope.

[0094] Data analysis: The experimental data were statistically analyzed using SPSS 16.0 software. The t-test was used for comparison between two groups, and one-way ANOVA was used for comparison among multiple groups, expressed as Mean±SEM. When P < 0.05, it was considered statistically significant.

[0095] The results are as Figure 3a and Figure 3b shown.

[0096] Example 3: Effects of mPGES-2 knockdown or overexpression on the expression of PCNA and Ki67 proteins

[0097] The expression levels of PCNA and Ki67 proteins in mouse hepatoma cells Hepa 1-6 NC, Hepa 1-6 KD, and Hepa 1-6 OE were detected by Western Blot.

[0098] The protein lysis method is described in "Example 1".

[0099] The experimental results are as Figure 4a and Figure 4b shown.

[0100] Example 4: Effects of mPGES-2 knockdown or overexpression on the cell cycle progression of hepatoma cells

[0101] Experimental method

[0102] Plate mPGES-2 NC, mPGES-2 KD, and mPGES-2 OE cells at a cell density of 3×10 5 / well / 2 mL (6-well plate) and culture in normal complete medium for 24 h. Harvest the cells, centrifuge at 1000 rpm for 5 min. Discard the supernatant, add 3 mL of ice-cold 70% ethanol while vortexing, and place at 4°C for more than 15 min. Add 10 mL of PBS containing 1% FBS to each tube, centrifuge at 1500 rpm for 5 min to obtain the precipitate. Add 300 μL of PI staining solution containing 1% RNase A, mix well, incubate at 37°C for 20 min, filter through a 200-mesh sieve, and analyze on a flow cytometer. The results are as Figure 5a and Figure 5b shown.

[0103] Example 5: Detection of mPGES-2 nuclear entry and chromatin binding in mouse hepatoma cells Hepal-6 by nuclear-cytoplasmic separation and chromatin extraction

[0104] a. Nuclear and cytoplasmic separation: Detected using a nuclear and cytoplasmic protein extraction kit (brand: Beyotime, P0027). Mouse hepatoma cells Hepa 1-6 were added to cytoplasmic protein extraction reagent A supplemented with PMSF. Vortex vigorously at the highest speed for 5 s to completely suspend and disperse the cell pellet. Incubate on ice for 10 - 15 min. Add cytoplasmic protein extraction reagent B. Vortex vigorously at the highest speed for 5 s and incubate on ice for 1 min. Vortex vigorously at the highest speed for 5 s and centrifuge at 12,000 - 16,000 g at 4°C for 5 min. Immediately aspirate the supernatant into a pre-cooled 1.5 mL EP tube, which is the extracted cytoplasmic protein. For the pellet, completely aspirate the remaining supernatant and add nuclear protein extraction reagent supplemented with PMSF. Vortex vigorously at the highest speed for 15 s to completely suspend and disperse the cell pellet. Then place it back on ice and vortex vigorously at high speed for 15 - 30 s every 1 - 2 min for a total of 30 min. Centrifuge at 12,000 - 16,000 g at 4°C for 10 min. Immediately aspirate the supernatant into a pre-cooled 1.5 mL EP tube, which is the extracted nuclear protein. Subsequently, WB was used to detect the nuclear entry of protein mPGES-2, and the results are as Figure 6a shown.

[0105] b. Chromatin extraction: Mouse hepatoma cells Hepa 1-6 were added with CSK-Triton reagent and lysed on ice for 10 min, vortexing once every 1 - 2 min, centrifuged at 1500 g for 5 min, the supernatant was aspirated, the pellet was gently pipetted with CSK again, and centrifuged at 1500 g for 5 min. The obtained pellet is the chromatin-bound protein. Subsequently, WB was used to detect the chromatin binding of protein mPGES-2. The results are as Figure 6b shown.

[0106] Experimental results show:

[0107] (1) Knockdown or overexpression of mPGES-2 inhibits or promotes the proliferation of hepatoma cells respectively: Uncontrolled cell proliferation is one of the main characteristics of hepatoma cells. To detect the effect of mPGES-2 on the proliferation of hepatoma cells, control (NC), mPGES-2 knockdown (KD), and mPGES-2 overexpression (OE) cell lines were constructed by lentivirus infection. Detection of mPGES-2 expression by Western blot showed that the cell lines were successfully constructed (as shown in Figure 1a and Figure 2a ). Crystal violet experiment showed that the colony formation of mPGES-2 KD was reduced compared with NC (***P < 0.001) (as shown in Figure 1b ), and the colony formation of mPGES-2 OE was increased compared with NC (**P < 0.01) (as shown in Figure 2bAs shown in the figure. The results of the CCK8 experiment showed that the cell proliferation of mPGES-2KD was slower compared with that of NC (*P < 0.05, **P < 0.01) (as Figure 1c shown), and the cell proliferation of mPGES-2OE was faster compared with that of NC (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001) (as Figure 2c shown). The above results indicated that knockdown of mPGES-2 inhibited the proliferation of liver cancer cells, and overexpression of mPGES-2 promoted the proliferation of liver cancer cells.

[0108] (2) Knockdown or overexpression of mPGES-2 inhibited or promoted DNA replication in liver cancer cells respectively: Efficient DNA replication is an important guarantee for the successful completion of cell proliferation. To detect the effect of mPGES-2 on DNA replication in liver cancer cells, the EdU Click it method was used to detect the DNA replication efficiency, and the EdU fluorescence intensity of PCNA-positive cells was statistically analyzed. The results showed that knockdown of mPGES-2 reduced the DNA replication efficiency of Hepa 1-6 cells (****P < 0.0001) (as Figure 3a shown), and overexpression of mPGES-2 increased the DNA replication efficiency of Hepa 1-6 cells (****P < 0.0001) (as Figure 3b shown). The above results indicated that knockdown of mPGES-2 inhibited DNA replication in liver cancer cells, and overexpression of mPGES-2 promoted DNA replication in liver cancer cells.

[0109] (3) mPGES-2 promoted the expression of cell proliferation antigen ki67 and DNA replication marker PCNA in liver cancer cells: Ki-67 is a marker protein for cell proliferation; PCNA is a marker protein for DNA replication. To further detect the effect of mPGES-2 on the proliferation and DNA replication of liver cancer cells, Western blot was used to detect the expression of ki67 and PCNA. The experimental results showed that knockdown of mPGES-2 downregulated the expression of ki67 and PCNA in Hepa 1-6 cells (*P < 0.05) (as Figure 4a shown), and overexpression of mPGES-2 increased the DNA replication efficiency of Hepa 1-6 cells (*P < 0.05) (as Figure 4b shown). This result further indicated that mPGES-2 promoted the proliferation and DNA replication of liver cancer cells.

[0110] (4) mPGES-2 interacted with Ki-67 in the nucleus of liver cancer cells to promote the proportion of S phase: The DNA replication process occurs in the S phase of the cell. Therefore, the effect of mPGES-2 on the S phase of the cell was detected, and the results showed that knockdown of mPGES-2 inhibited the proportion of the S phase of the cell (as Figure 5aAs shown, overexpression of mPGES-2 increased the proportion of cells in the S phase (as Figure 5b shown). This indicates that the interaction between mPGES-2 and Ki67 regulates DNA replication by affecting the S phase of cells.

[0111] (5) mPGES-2 can enter the nucleus and bind to chromatin in liver cancer cells: The localization of proteins in cells is closely related to their functions. The experimental results showed that mPGES-2 can affect DNA replication in liver cancer cells. Therefore, it was detected whether mPGES-2 can enter the nucleus. Using the nuclear-cytoplasmic separation experiment, it was found that mPGES-2 can enter the nucleus in liver cancer cells (cy represents cytoplasm; Nu represents nucleus) (as Figure 6a shown). Further, through the chromatin extraction experiment, it was found that after entering the nucleus in liver cancer cells, mPGES-2 binds to chromatin (S represents proteins not binding to chromatin; C represents chromatin-binding proteins) (as Figure 6b shown).

[0112] In summary, through the above experimental comparisons, this study first proposed the regulatory effect of mPGES-2 on liver cancer cells, found that inhibiting mPGES-2 can inhibit DNA replication and proliferation of liver cancer cells, while overexpressing mPGES-2 can promote DNA replication and proliferation of liver cancer cells, and in tumor cells, mPGES-2 can enter the nucleus and bind to chromatin. It provides potential molecular targets for liver cancer treatment regimens based on DNA replication.

[0113] In addition, the inventor of this case also referred to the foregoing embodiments and conducted tests with other raw materials, process operations, and process conditions described in this specification, and all obtained relatively ideal results.

[0114] The above shows and describes the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. What is described in the above embodiments and the specification only illustrates the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed.

Claims

1. Application of mPGES-2 as a target in the preparation of drugs for preventing and / or treating liver cancer.

2. The use according to claim 1, characterized in that: The target knockdown can at least inhibit the proliferation of liver cancer cells in liver cancer mice; and / or, the target knockdown can at least inhibit DNA replication of liver cancer cells in liver cancer mice; And / or, the target knockdown can at least downregulate the expression of Ki67 and PCNA in liver cancer cells of liver cancer mice.

3. The use according to claim 1, characterized in that: Overexpression of the target can at least promote the proliferation of liver cancer cells in liver cancer mice; And / or, the overexpression of the target can at least promote DNA replication of liver cancer cells in liver cancer mice; And / or, the overexpression of the target can at least upregulate the expression of Ki67 and PCNA in liver cancer cells of liver cancer mice.

4. The use according to claim 1, characterized in that: The target knockdown can at least reduce the proportion of liver cancer cells in the S phase in liver cancer mice.

5. The use according to claim 1, characterized in that: Overexpression of the target can at least increase the proportion of liver cancer cells in the S phase in liver cancer mice.

6. The use according to claim 1, characterized in that: The mPGES-2 can at least enter the nucleus of liver cancer cells and bind to chromatin.

7. Use of mPGES-2 or an inhibitor of its encoding gene in the preparation of a drug for preventing and / or treating liver cancer.

8. The use according to claim 7, characterized in that: The inhibitor is selected from interfering molecules that specifically interfere with the expression of the gene encoding mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its encoding gene.

9. A pharmaceutical composition for preventing and / or treating liver cancer, characterized in that: include: An inhibitor of mPGES-2 or its encoding gene, and a pharmaceutically acceptable carrier.

10. The pharmaceutical composition according to claim 9, characterized in that: The inhibitor is selected from interfering molecules that specifically interfere with the expression of the gene encoding mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its encoding gene.

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