siRNA targeting PABPC4 and its application in the treatment of liver cancer

Abstract

The present invention discloses siRNA targeting PABPC4 and its application in the treatment of liver cancer, belonging to the field of biomedicine. It is determined that the expression level of PABPC4 is significantly correlated with the nuclear positivity of MYC, a series of siRNA targeting PABPC4 are screened, and after transfection of liver cancer cells, it is confirmed that the use of siRNA to target and inhibit the expression of ABPC4 can significantly inhibit the proliferation and cloning ability of liver cancer cells, and can significantly inhibit the high expression of MYC and PABPC4, which is of great significance for the development of new liver cancer gene drugs against high expression of MYC and PABPC4 and improving the treatment effect of liver cancer with high expression of MYC and PABPC4, and has significant application prospects and economic value.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to siRNA targeting PABPC4 and application thereof in the treatment of liver cancer. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Abnormal activation of the transcription factor MYC can lead to the occurrence of a variety of tumors; among them, more than 30% of hepatocellular carcinomas (HCC) show abnormal activation of the oncogene MYC; however, the development of small molecule inhibitors that specifically target MYC has not been successful so far. Since dysregulation of gene expression caused by MYC activation is a key driver of tumor formation and may be dependent on specific transcriptional programs, in-depth exploration of the key molecules and pathways associated with MYC in the occurrence and development of liver cancer and the search for synergistic lethal strategies associated with abnormal MYC activation are new ideas for achieving MYC-specific targeted treatment of liver cancer.

[0004] The poly(A)-tail at the 3' end of most eukaryotic mRNAs plays a key role in their stability, nuclear transport, and translation. These effects are largely mediated by poly(A) binding proteins (PABPs), which are involved in RNA biogenesis and transcription-translation functions. Five PABP proteins are co-expressed in human cells, PABP1, PABPC3, PABP5, PABPC1L, and iPABP. iPABP (also known as PABPC4) is a protein that is upregulated in activated T cells and is also expressed on the surface of activated platelets, and its mRNA is also expressed in other tissues. PABPC4 protein is highly expressed in a variety of tumors, including colon cancer, lung adenocarcinoma, triple-negative breast cancer, and hepatocellular carcinoma. Studies have shown that PABPC4 can promote the growth and metastasis of triple-negative breast cancer, but the mechanism of action of PABPC4 in HCC is currently little explored. Summary of the invention

[0005] In order to solve the deficiencies of the prior art, the purpose of the present invention is to provide siRNA targeting PABPC4 and its application in the treatment of liver cancer. The present invention is applied to the treatment of liver cancer by effectively inhibiting the expression level of PABPC4.

[0006] In order to achieve the above object, the technical solution of the present invention is:

[0007] In order to confirm the correlation between PABPC4 protein level and MYC nuclear staining, the present invention performed immunohistochemical staining on tumor samples of 85 liver cancer patients, and determined that the PABPC4 expression level was significantly positively correlated with the nuclear positivity of MYC; secondly, a series of siRNAs targeting PABPC4 were screened, and after transfection of liver cancer cells, the efficiency of siRNA silencing PABPC4 was detected by fluorescent quantitative PCR. At the same time, immunohistochemical staining, CCK-8, Western blot and other experimental methods were used to detect the effects of siRNA on the proliferation and cloning of liver cancer cells. The results showed that multiple siRNA transfection had a high silencing efficiency on PABPC4, and could significantly inhibit the proliferation and cloning ability of liver cancer cells with high expression of MYC and PABPC4, that is, significantly inhibited the progression of MYC high-activity molecular subtype liver cancer.

[0008] In a first aspect of the present invention, the present invention provides siRNA capable of effectively inhibiting the PABPC4 gene, wherein the sequence of the siRNA is selected from one or more of SEQ ID NO. 1-2 or 3-4 or 5-6.

[0009] The siRNA provided by the invention reduces the expression level of the PABPC4 gene in liver cancer cells by more than 50%.

[0010] Preferably, when the sequence of siRNA is SEQ ID NO. 3-4, the effect of inhibiting the expression of PABPC4 gene is better.

[0011] The second aspect of the present invention provides a use of siRNA targeting PABPC4 gene in the preparation of a drug for treating liver cancer, wherein the sequence of the siRNA is selected from one or more of SEQ ID NO. 1-2 or 3-4 or 5-6.

[0012] Preferably, the sequence of the siRNA is selected from SEQ ID NO.3-4.

[0013] Preferably, the siRNA targeting the PABPC4 gene achieves the purpose of treating liver cancer by inhibiting the proliferation and cloning of liver cancer cells.

[0014] Preferably, the liver cancer is a MYC high activity molecule subtype liver cancer.

[0015] The third aspect of the invention provides a pharmaceutical preparation for treating liver cancer, the pharmaceutical preparation comprising siRNA or a nucleic acid sequence modification thereof and a carrier, wherein the siRNA or a nucleic acid sequence modification thereof inhibits the expression of the PABPC4 gene;

[0016] The sequence of the siRNA is selected from one or more of SEQ ID NO. 1-2 or 3-4 or 5-6.

[0017] In the pharmaceutical preparation for treating liver cancer (eg HCC) provided by the present invention, the nucleic acid sequence modification product is obtained by performing one or more modifications of ribose modification, base modification and phosphate backbone modification on any nucleotide of the siRNA.

[0018] Preferably, the carrier is selected from viruses, nanoparticles, cholesterol or liposomes.

[0019] Preferably, the liver cancer is a MYC high activity molecule subtype liver cancer.

[0020] The beneficial effects of the present invention are:

[0021] The present invention reduces the expression level of PABPC4 by siRNA targeting the PABPC4 gene, which can effectively inhibit the proliferation and clone formation ability of liver cancer cells. It is of great significance for developing new anti-MYC and PABPC4 high-expression liver cancer gene drugs and improving the treatment effect of liver cancer with high expression of MYC and PABPC4, and has significant application prospects and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0023] Figure 1 is the detection result of the embodiment of the present invention;

[0024] Among them, A is the representative result of MYC and PABPC4 immunohistochemical staining in patients with liver cancer;

[0025] B is a quantitative analysis of the immunohistochemical MYC and PABPC4 expression in A. The expression of MYC and PABPC4 in liver cancer is positively correlated;

[0026] C is the correlation between the expression levels of PABPC4 and MYC in different liver cancer cell lines;

[0027] D shows the different knockdown efficiencies of three PABPC4 siRNAs in the cell line SK-Hep1 that highly expresses PABPC4;

[0028] E is the effect of three siRNA knockdowns on liver cancer cell proliferation detected by CCK8. The knockdown effect of PABPC4 is related to the inhibition of cell proliferation. In contrast, MYC low-expressing cell lines SNU-387, SNU-182 and Li7 were not affected.

[0029] F and G are crystal violet staining images showing that PABPC4 knockdown reduced the clonal formation ability of HCC cell lines in the clone formation experiment, while MYC low-expressing cell lines SNU-387, SNU-182 and Li7 cells were almost unaffected. DETAILED DESCRIPTION

[0030] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0031] Example 1

[0032] 1. Experimental Materials

[0033] 1. Experimental cell lines

[0034] SK-Hep1: human hepatocellular carcinoma tumor cells, obtained from the cell bank of the Chinese Academy of Sciences, cultured in DMEM supplemented with 10% FBS and 1% double antibodies.

[0035] HepG2: human hepatocellular carcinoma tumor cells, obtained from the cell bank of the Chinese Academy of Sciences, cultured in DMEM supplemented with 10% FBS and 1% double antibodies.

[0036] SNU-387: human hepatocellular carcinoma cells, purchased from Procell cell bank, cultured in DMEM supplemented with 10% FBS and 1% double antibodies.

[0037] SNU-182: human liver cancer tumor cells, purchased from Procell cell bank, cultured in DMEM supplemented with 10% FBS and 1% double antibodies.

[0038] Li7: Human hepatocellular carcinoma cells, obtained from the cell bank of the Chinese Academy of Sciences, cultured in DMEM supplemented with 10% FBS and 1% double antibodies.

[0039] 2. Experimental Reagents

[0040] (1) Cell culture medium: DMED culture medium was purchased from MyChen Technology (Beijing) Co., Ltd.

[0041] (2) Fetal bovine serum: purchased from Gibco.

[0042] (3) PBS powder: purchased from Wuhan Saiweier Biotechnology Co., Ltd.

[0043] (4) 0.25% pancreatic enzyme solution: purchased from Mychen Technology (Beijing) Co., Ltd.

[0044] (5) Small interference was purchased from Sangon Biotechnology (Shanghai) Co., Ltd.

[0045] (6) PABPC4 antibody (catalog number DF9762) was purchased from Affinity Bioscience; GAPDH antibody (catalog number AB0036) was purchased from Abways Technology; rabbit secondary antibody (catalog number AB0101) was purchased from Abways Technology; immunohistochemistry secondary antibody (catalog number PV-9000) was purchased from Beijing Sino-Golden Bridge Biotechnology Co., Ltd.

[0046] (7) Immunohistochemistry DAB staining solution was purchased from Zhongshan Jinqiao Biotechnology (Beijing) Co., Ltd.

[0047] (8) CCK8, catalog number CA1210, was purchased from Solebow Technology (Beijing) Co., Ltd.

[0048] (9) 6 cm culture dishes, 6-well culture dishes, 12-well culture dishes, and 96-well culture dishes were purchased from Jie Te Biofiltration (Guangzhou) Co., Ltd.

[0049] (10) Crystal violet dye (catalog number C8470) was purchased from Solebow Technology (Beijing) Co., Ltd.

[0050] 3. Main experimental instruments

[0051]

[0052]

[0053] 2. Experimental Methods

[0054] 1. Cell culture and passaging

[0055] Cells were cultured in 6 cm culture dishes, 5 mL of culture medium was added to each dish, and when the cell density reached 80%, the culture medium was discarded and the cells were washed once with PBS.

[0056] Add 1 mL of 0.25% trypsin and digest for 2 min at room temperature.

[0057] Observe under a microscope. When the cells become round and bright, discard the trypsin and add 2 mL of culture medium to terminate digestion.

[0058] Gently pipette the cells off the culture dish and transfer the cell suspension to a 5 mL centrifuge tube.

[0059] Centrifuge at 1000 pm for 5 min, discard the supernatant, flick off the cell pellet, and suspend the cell pellet in 1 mL of culture medium.

[0060] The cell suspension was inoculated into a new culture dish, 5 mL of culture medium was added, and cultured in a 37°C constant temperature incubator.

[0061] 2. Cell Transfection

[0062] According to the instructions, siRNA-1, siRNA-2 and siRNA-3 were prepared into 20 μM solutions using DEPC water for later use.

[0063] After digestion, the cells were plated in 12-well plates and transfected when the cell density reached 30-50% confluence.

[0064] Prepare five sterile 1.5mL EP tubes, add 50μL of OPTI-MEM and 2μL of the corresponding siRNA-1, siRNA-2, siRNA-3 and negative control siRNA solution to four EP tubes respectively. Flick for 3s to mix thoroughly; then add 200μL of OPTI-MEM and 12μL of lipo2000 transfection reagent to the remaining EP tube respectively, let stand at room temperature for 5 minutes, add OPTI-MEM mixed with lipo2000 transfection reagent to form a transfection complex between siRNA and transfection reagent, then add to the corresponding wells of the 12-well plate, and shake gently to mix well.

[0065] 3. Total RNA Extraction

[0066] Total RNA of tissues was extracted using Trizol reagent (Nanjing Novozyme Co., Ltd.) according to its instructions. The specific steps are as follows:

[0067] Every 5×10 6 -1×10 7 Add 1 mL RNA isolater to each cell. Pipette repeatedly until cells are fully lysed and place on ice for 5 min.

[0068] Add about 1 / 5 volume of chloroform, mix thoroughly by inverting for about 1 min, and let stand at room temperature for 5 min.

[0069] Centrifuge at 12,000 rpm for 15 min at 4°C.

[0070] Carefully remove the supernatant, avoiding touching the middle layer, transfer the supernatant into a new 1.5 mL centrifuge tube, add an equal volume of isopropanol, gently invert to mix, and let stand at room temperature for 5 min.

[0071] Centrifuge at 12,000 rpm for 15 min at 4°C.

[0072] The supernatant was removed by aspiration, and the precipitate was retained. 1 mL of 75% ethanol was added to the precipitate, and the precipitate was washed by centrifugation at 4°C and 12,000 rpm for 15 min.

[0073] Aspirate the supernatant, let the precipitate dry naturally at room temperature, add an appropriate amount of RNase-free water, and use the Tip to blow and suck to fully dissolve the precipitate.

[0074] Take 1-2 μL and measure RNA concentration and A260 / A280 value. Generally, the A260 / A280 ratio is 1.8-2.1. Store RNA in a -80℃ refrigerator until use.

[0075] 4. Reverse transcription and cDNA synthesis

[0076] cDNA was reverse transcribed using HiScript II Q Select RT SuperMix for qPCR (+gDNA wiper) (Nanjing Novozymes Corporation), with a total reaction system (total volume of 10 μL) and a total reverse transcribed RNA volume of 500 ng.

[0077] 5.RT-PCR

[0078] Instrument used: Real-time fluorescence quantitative PCR instrument (Roche LC96) Quantitative Real-time PCR (LightCycler 96).

[0079] Template: The above reverse transcription product was diluted at a ratio of 1:10 and used as a template for PCR reaction.

[0080] Primer sequences qPCR verification primer sequences are as follows:

[0081] PABPC4:

[0082] Sense sequence: 5′-AAGCCAATCCGCATCATGTG-3′ (SEQ ID NO. 7)

[0083] Antisense sequence: 5′-CTCTTGGGTCTCGAAGTGGAC-3′ (SEQ ID NO. 8)

[0084] GAPDH:

[0085] Sense sequence: 5′-ACAACTTTGGTATCGTGGAAGG-3′ (SEQ ID NO. 9)

[0086] Antisense sequence: 5′-GCCATCACGCCACAGTTTC-3′ (SEQ ID NO. 10)

[0087] RT-PCR reaction system:

[0088] The reaction used 2X Universal SYBR Green Fast qPCRMix, product number RK21203, from Wuhan Abotek Biotechnology Co., Ltd. The reagents and their dosages were as follows:

[0089] 2X Universal SYBR Green Fast qPCR Mix 10μL

[0090] DNA template 4.5 μL

[0091] Forward primer (10 μM) 0.4 μL

[0092] Reverse primer (10 μM) 0.4 μL

[0093] ddH2O to 20μL

[0094] The reaction procedure was as follows: pre-denaturation at 95°C for 3 min, and cyclic reaction at 95°C for 5 s for 40-45 cycles.

[0095] The results were analyzed using GraphpadPrism8 statistical software package, and quantitative comparisons were analyzed using t-test.

[0096] 6. Immunohistochemical Staining

[0097] The immunohistochemical sections were treated with xylene for 10 min three times, anhydrous ethanol for 5 min twice, 95% ethanol for 5 min once, and then soaked in water for 5 min.

[0098] Immerse the slices in sodium citrate repair solution or EDTA pH 9.0 repair solution and microwave boiling repair for 10 minutes.

[0099] The sections were treated with hydrogen peroxide for 10 min, perforated with 1% triton for 10 min, and blocked with 5% BSA for 30 min.

[0100] After incubation with primary antibody at 4°C overnight, the sections were washed with PBS three times, 5 min each time.

[0101] After incubation with the secondary antibody for 1.5 h at room temperature, the sections were washed three times with PBS and developed with DAB.

[0102] The immunohistochemical results were analyzed by Olympus panoramic scanning microscope.

[0103] 7.CCK8 cell activity detection

[0104] The cell suspension was inoculated in a 96-well plate (100 μL / well), and the culture plate was placed in an incubator for pre-culture (under the conditions of 37° C., 5% CO 2 ).

[0105] Add 10 μL of CCK-8 solution and 90 μL of complete culture medium to each well (be careful not to create bubbles in the wells, as they will affect the OD reading).

[0106] Incubate the plate in an incubator for 1-4 hours.

[0107] The absorbance at 450 nm was measured using an enzyme reader.

[0108] 8. Colony formation and statistics

[0109] SK-Hep1, HepG2, HuH7, HuH6, Li7, SNU-387, and SNU-182 cell lines in the logarithmic growth phase were digested with trypsin and plated into 96-well plates at a density of 5000 cells per well.

[0110] 16 hours later, 5 nM siRNA-1, siRNA-2, siRNA-3 or control small interfering RNA were transfected, and a second transfection with the same amount was performed 24 hours after transfection.

[0111] 48 hours after the second transfection, the cells were completely digested with trypsin and evenly plated into 6-well plates.

[0112] After 2 to 3 weeks of culture, the cells were fixed, stained with crystal violet, and photographed.

[0113] The clone formation images were analyzed by Image pro plus software.

[0114] 9.Western blot

[0115] After the protein was lysed using 1x loading buffer, the protein sample was first electrophoresed using 10% SDS-polyacrylamide gel (SDS-PAGE).

[0116] The proteins were transferred to the NC membrane by electrotransfer (constant current 200 mA, 4°C electrotransfer for 1.5 h).

[0117] The transferred NC membrane was blocked for 60 min using TBST buffer (pH 7.4, 10 mM Tris-HCl, 0.9% NaCl) containing 5% skim milk powder as a blocking agent.

[0118] The membrane was immersed in the primary antibody dilution solution and shaken overnight at 4°C in a shaker to allow the diluted antibody to fully bind to the NC membrane.

[0119] The membrane was washed three times with TBST buffer, 10 min each time.

[0120] Goat anti-rabbit IgG (diluted 2000 times) was used as the secondary antibody and added to 5% skim milk powder blocking solution, and the membrane was immersed in it and continued to shake on a shaker at room temperature for 1 hour. The membrane was washed 4 times with TBST buffer.

[0121] Finally, ECL luminescent liquid was used for color development.

[0122] IV. Experimental Results

[0123] 1. Immunohistochemical staining to obtain the expression of PABPC4 gene in liver cancer patient tissues

[0124] To confirm the correlation between PABPC4 protein levels and MYC nuclear staining, immunohistochemical staining was performed on tumor samples from 85 HCC patients. Figure 1 A shows immunohistochemical staining of MYC and PABPC4 in patients with liver cancer. Figure 1 B is the quantitative analysis of MYC and PABPC4 expression by immunohistochemistry in Figure A. Figure 1 In B, it can be determined that the expression level of PABPC4 is significantly positively correlated with the nuclear positivity of MYC, that is, the expression of MYC and PABPC4 in liver cancer is positively correlated.

[0125] 2. Western blot detection of the correlation between PABPC4 protein level and MYC protein level in different liver cancer cell lines

[0126] In order to confirm the correlation between PABPC4 protein level and MYC protein level, WB experiments were performed on 7 commonly used liver cancer cell lines. Figure 1 C shows the WB experimental results and quantitative analysis. Figure 1 In C, it can be determined that the expression level of PABPC4 is significantly correlated with the expression level of MYC, that is, the expression of PABPC4 is higher in the liver cancer cell line with high MYC expression.

[0127] 3. Selection of siPABPC4 target sequence

[0128] Three siRNA sequences were designed for the human PABPC4 gene and synthesized by Sangon Biotech (Shanghai) Co., Ltd.

[0129] The base sequence of siRNA-1 is as follows,

[0130] Sense sequence: 5′-GUGUCAAGGUGAUGAGAGATT-3′ (SEQ ID NO. 1)

[0131] Antisense sequence: 5′-UCUCUCAUCACCUUGACACTT-3′ (SEQ ID NO. 2)

[0132] The base sequence of siRNA-2 is as follows:

[0133] Sense sequence: 5′-GGAGAGAAUUAGUCGAUAUTT-3′ (SEQ ID NO. 3)

[0134] Antisense sequence: 5′-AUAUCGACUAAUUCUCUCUCCTT-3′ (SEQ ID NO. 4)

[0135] The base sequence of siRNA-3 is as follows:

[0136] Sense sequence: 5′-GAACUUGGAUGACACUAUUTT-3′ (SEQ ID NO. 5)

[0137] Antisense sequence: 5′-AAUAGUGUCAUCCAAGUUCTT-3′ (SEQ ID NO.6)

[0138] 4. RT-PCR detection of PABPC4 siRNA silencing effect

[0139] Cell culture conditions and culture medium: SK-Hep1 cell lines were cultured in DMEM containing 10% fetal bovine serum in a 5% CO2, 37°C incubator. After the cells were plated into 12-well plates, 2 μL of small interfering RNA and 4 μL of lipo2000 transfection reagent were used in each well to knock down PABPC4 expression, and cell RNA was collected 48 hours after transfection. RNA was extracted with Trizol and then reverse transcribed, and qPCR was used to verify the knockout efficiency of PABPC4. The primer sequences for PABPC4 qPCR verification are as follows:

[0140] Sense sequence: 5′-AAGCCAATCCGCATCATGTG-3′ (SEQ ID NO. 7)

[0141] Antisense sequence: 5′-CTCTTGGGTCTCGAAGTGGAC-3′ (SEQ ID NO. 8)

[0142] like Figure 1 As can be seen from the left figure in D, the q-PCR results show that all three siRNAs can knock down the PABPC4 mRNA level in the cell line SK-Hep1 that highly expresses PABPC4, among which small interfering RNA No. 2 has the best knockdown effect.

[0143] 4. WB detection of PABPC4 siRNA silencing effect

[0144] SK-Hep1 liver cancer cell lines in the logarithmic growth phase were digested with trypsin and seeded in 12-well plates. After growing to a density of 50% to 60%, they were transfected with 40 nM siRNA-1, siRNA-2, siRNA-3 or control small interfering proteins, and the proteins were collected using 1x Loading buffer 48 h after transfection. Figure 1 As can be seen from the right figure in D, WB results show that all three siRNAs can knock down the expression level of PABPC4 protein in the cell line SK-Hep1 that highly expresses PABPC4, among which small interfering RNA No. 2 has the best knockdown effect.

[0145] 5. CCK8 analysis of the effect of PABPC4 siRNA on the growth of hepatoma cell lines

[0146] SK-Hep1, HepG2, HuH7, HuH6, Li7, SNU-387, and SNU-182 cell lines in the logarithmic growth phase were digested with trypsin and plated into 96-well plates at a density of 5,000 cells per well. After 16 hours, 5 nM siRNA-1, siRNA-2, siRNA-3, or control small interfering RNA were transfected, respectively. CCK-8 staining and UV spectrophotometer readings were performed 72 hours after transfection. Figure 1 As shown in E, knocking down the expression of PABPC4 had a significant inhibitory effect on the proliferation ability of 4 cell lines with high MYC and PABPC4 expression levels, namely SK-Hep1, HepG2, HuH7 and HuH6, among which the best inhibitory effect was achieved by small interference No. 2; while it had no effect on the proliferation ability of Li7, SNU-387 and SNU-182 cell lines with low MYC and PABPC4 expression levels.

[0147] 6. Detection of the effect of siPABPC4 on cell clone formation

[0148] SK-Hep1, HepG2, HuH7, HuH6, Li7, SNU-387, and SNU-182 cell lines in the logarithmic growth phase were digested with trypsin and plated into 96-well plates at a density of 5000 per well. After 16 hours, 5nM siRNA-1, siRNA-2, siRNA-3 or control small interference were transfected respectively. The same amount of secondary transfection was performed 24 hours after transfection. After the second transfection, the cells were completely digested with trypsin and evenly plated into 6-well plates 48 hours later. After culturing for 2 to 3 weeks, the clone formation was subjected to crystal violet staining. Figure 1 F. Figure 1 As shown in G, knocking down the expression of PABPC4 had a significant inhibitory effect on the cloning ability of 4 cell lines with high MYC and PABPC4 expression levels, namely SK-Hep1, HepG2, HuH7 and HuH6, among which the best inhibitory effect was achieved by small interference No. 2; however, it had no effect on the cloning ability of Li7, SNU-387 and SNU-182 cell lines with low MYC and PABPC4 expression levels.

[0149] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A siRNA, characterized in that: This siRNA inhibits the expression of the PABPC4 gene; The sequence of the siRNA is selected from one or more of SEQ ID NO. 1-2 or 3-4 or 5-6.

2. The siRNA according to claim 1, characterized in that: The sequence of the siRNA is SEQ ID NO.3-4.

3. Use of siRNA targeting PABPC4 gene in the preparation of a drug for treating liver cancer, wherein the sequence of the siRNA is selected from one or more of SEQ ID NO. 1-2 or 3-4 or 5-6.

4. The use according to claim 3, characterized in that: The sequence of the siRNA is selected from SEQ ID NO.3-4.

5. The use according to claim 3, characterized in that: The liver cancer is a MYC high activity molecule subtype liver cancer.

6. A pharmaceutical preparation for treating liver cancer, characterized in that: The pharmaceutical preparation comprises siRNA or a nucleic acid sequence modification thereof and a carrier, wherein the siRNA or the nucleic acid sequence modification thereof inhibits the expression of the PABPC4 gene; The sequence of the siRNA is selected from one or more of SEQ ID NO. 1-2 or 3-4 or 5-6.

7. The pharmaceutical preparation according to claim 6, characterized in that: The nucleic acid sequence modification is obtained by performing one or more modifications of ribose modification, base modification and phosphate backbone modification on any nucleotide of the siRNA; The carrier is selected from viruses, nanoparticles, cholesterol or liposomes.

8. The pharmaceutical preparation according to claim 6, characterized in that: The liver cancer is a MYC high activity molecule subtype liver cancer.

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