Application of inhibitors targeting lncRNA CTC-490G23.2 in the preparation of anti-tumor drugs

Abstract

The present invention discloses the use of an inhibitor targeting lncRNA CTC‑490G23.2 in the preparation of an anti-tumor drug, and belongs to the field of biomedicine technology. The inhibitor is an antisense oligonucleotide delivery nanoparticle constructed based on the cell-penetrating peptide LAH4‑L1, and the effective concentration of the antisense oligonucleotide delivery nanoparticle is 0.25‑0.5 mg / kg. The present invention first discovered that targeted inhibition of lncRNA CTC‑490G23.2 expression can inhibit the invasion and metastasis of esophageal cancer, specifically by targeting and inhibiting the expression of lncRNA CTC‑490G23.2. Antisense oligonucleotide delivery nanoparticles provided by the present invention for inhibiting the expression of lncRNA CTC‑490G23.2 have certain safety as a potential anti-tumor drug.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of an inhibitor targeting lncRNA CTC-490G23.2 in the preparation of anti-tumor drugs. Background Art

[0002] Esophageal cancer is the sixth leading cause of cancer death in the world. It is a complex disease with causes that vary by histological type and population. Its metastasis accounts for approximately 90% of cancer deaths worldwide, and both morbidity and mortality are among the top ten in China. The causes of ESCC are diverse and vary in different regions. Esophageal cancer is divided into esophageal squamous cell carcinoma (ESCC) and adenocarcinoma (EADC). Among them, ESCC is one of the most common malignant tumors in my country with a high mortality rate, but its mechanism has not been adequately studied and remains to be elucidated. ESCC currently still lacks effective drugs for the treatment of patients with metastatic cancer (PMID: 28823862).

[0003] Long noncoding RNAs (lncRNAs) are a group of heterogeneous non-protein coding transcripts with a length of more than 200 nucleotides. Increasing evidence shows that lncRNAs, as oncogenes or tumor suppressors, play a complex and precise regulatory role in the occurrence and development of cancer. lncRNAs participate in many physiological and pathological processes such as cell proliferation, immunity, and metastasis through epigenetic, transcriptional, and translational regulation. Previous studies have found that NAT10-mediated ac4C modification leads to the stabilization and overexpression of lncRNA CTC-490G23.2 in primary esophageal squamous cell carcinoma (ESCC) and its further upregulation in metastatic tissues, making lncRNA CTC-490G23.2 significantly promote tumor invasion and metastasis both in vivo and in vitro. This means that targeting lncRNA CTC-490G23.2 has the potential to treat tumor metastasis.

[0004] Oligonucleotide technology is one of the mature technologies of targeted nucleotides. Oligonucleotide technology has been developed to specifically downregulate, upregulate or modify the expression of key genes in advanced diseases such as cancer or genetic diseases. It broadens the range of potential therapeutic targets. By controlling the expression of RNA interference proteins, proteins that were previously unaffected by traditional small molecules can be affected. Among them, antisense oligonucleotides (ASOs) targeting lncRNA have been developed and utilized, and more and more ASO-based treatments are being tested in clinical trials. Summary of the invention

[0005] The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide an application of an inhibitor targeting lncRNA CTC-490G23.2 in the preparation of an anti-esophageal cancer drug.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] Application of inhibitors targeting lncRNA CTC-490G23.2 in the preparation of anti-esophageal cancer drugs.

[0008] The full length sequence of the lncRNA CTC-490G23.2 is as follows (also shown in SEQ ID NO.1):

[0009] 5'--3'.

[0010] Furthermore, the inhibitor is any one or more combinations of siRNA, shRNA, antisense oligonucleotide, miRNA or ribozyme.

[0011] Furthermore, the inhibitor is an antisense oligonucleotide, comprising or consisting of the following sequence: 5'-GGAGAGGAGCTTAGCUCCAG-3', full-chain phosphorothioate modification + two-terminal 2'OME modification.

[0012] Furthermore, the inhibitor is an antisense oligonucleotide delivery nanoparticle constructed based on cell penetrating peptide (CPP).

[0013] Furthermore, the cell-penetrating peptide comprises the following sequence or consists of the following sequence: PKKKRKV-GCG-KKALLAHALHLLALLALHLAHALKKA; wherein PKKKRKV is a nuclear localization signal sequence, GCG is a linker sequence, and KKALLAHALHLLALLALHLAHALKKA is a LAH4-L1 peptide sequence.

[0014] Furthermore, the antisense oligonucleotide delivery nanoparticles are prepared by the following method: the cell penetrating peptide and the antisense oligonucleotide are mixed in nuclease-free water and vortexed to self-assemble to form the antisense oligonucleotide delivery nanoparticles.

[0015] Furthermore, the weight ratio of the cell penetrating peptide to the antisense oligonucleotide is 20:1.

[0016] Furthermore, the effective concentration of the antisense oligonucleotide delivery nanoparticles is 0.25-0.5 mg / kg.

[0017] Furthermore, the anti-esophageal cancer drug is a drug that can inhibit the invasion and / or metastasis of esophageal cancer cells and can significantly inhibit the invasion and metastasis of esophageal cancer cells.

[0018] Furthermore, the anti-esophageal cancer drug is a drug for treating and / or preventing esophageal cancer.

[0019] Furthermore, the esophageal cancer is esophageal squamous cell carcinoma.

[0020] An anti-esophageal cancer drug is the inhibitor described in the above application.

[0021] Furthermore, the anti-esophageal cancer drug also contains pharmaceutically acceptable excipients.

[0022] Furthermore, the pharmaceutically acceptable excipient is at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant and a lubricant.

[0023] Furthermore, the administration method of the anti-esophageal cancer drug includes but is not limited to tail vein injection.

[0024] Compared with the prior art, the present invention has the following advantages and effects:

[0025] The present invention discovers for the first time that targeted inhibition of lncRNA CTC-490G23.2 expression can inhibit the invasion and metastasis of esophageal cancer, specifically inhibiting the invasion and metastasis of esophageal cancer by delivering antisense oligonucleotides that target and inhibit the expression of lncRNA CTC-490G23.2.

[0026] The antisense oligonucleotide delivery nanoparticles targeted to inhibit the expression of lncRNA CTC-490G23.2 provided by the present invention have certain safety as a potential anti-tumor drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is the structure diagram of the SV40-LAH4-L1 peptide.

[0028] Figure 2 This is a graph showing the research results on the effect of lncRNA CTC-490G23.2 on the invasion ability of esophageal squamous cell carcinoma cells.

[0029] Figure 3 This is a graph showing the research results on the effect of lncRNA CTC-490G23.2 on the lymph node metastasis ability of esophageal squamous cell carcinoma cells.

[0030] Figure 4 This is a graph showing the research results on the effects of different ASO treatments targeting lncRNA CTC-490G23.2 on the invasion ability of esophageal squamous cell carcinoma cells.

[0031] Figure 5 This is a graph showing the effects of different ASO treatments targeting lncRNA CTC-490G23.2 on the expression of lncRNA CTC-490G23.2.

[0032] Figure 6 Polydispersity index (PDI) and zeta potential plots for the ratio of ASO and SV40-LAH4-L1 peptide targeting lncRNA CTC-490G23.2.

[0033] Figure 7 Gel image showing the optimal ratio of ASO and SV40-LAH4-L1 peptide targeting lncRNA CTC-490G23.2.

[0034] Figure 8 This is the immunofluorescence result of different nanodrugs entering the cell nucleus.

[0035] Fig. 9This is a graph showing the results of a study on the effect of ASO / SV40-LAH4-L1 peptide nanodrug targeting lncRNA CTC-490G23.2 on the invasion ability of esophageal cancer cells in vitro.

[0036] Fig.10 The fluorescence detection results of the lung metastasis ability of esophageal squamous cell carcinoma cells using different concentrations of ASO / SV40-LAH4-L1 peptide nanomedicine targeting lncRNA CTC-490G23.2 and its control.

[0037] Fig.11 The results of the morphological changes in the liver, lung, kidney, heart, spleen and other tissues of nude mice treated with different concentrations of ASO / SV40-LAH4-L1 peptide nanodrugs targeting lncRNA CTC-490G23.2. DETAILED DESCRIPTION

[0038] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0039] Human esophageal squamous cell carcinoma cells KYSE150 and KYSE410 were purchased from German Microbiology and Cell Culture Co., Ltd.; LB medium was purchased from Shanghai Bioengineering Co., Ltd., and 0.25% trypsin was purchased from Gibco, USA. Experimental female mice (NOD-SCID) were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

[0040] Human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC have been published in the literature "Wen WenXu et al. Direct Targeting of CREB1 with Imperatorin Inhibits TGFβ2-ERK Signaling toSuppressEsophageal Cancer Metastasis, Advanced Science, 2020. DOI: 10.1002 / advs.202000925". Human esophageal squamous cell carcinoma luciferase-labeled cells KYSE410-luc have been published in the literature "Hui-FangHu et al. Identification of miR-515-3p and its targets, vimentin and MMP3, as a key regulatory mechanism in esophageal cancer metastasis: functional and clinical significance".

[0041] KYSE150-LUC-LM3 cell construction method: 0.25% trypsin was used to digest the human esophageal squamous cell carcinoma luciferase-labeled KYSE150-LUC cells into single cells. 6 The LUCI-labeled KYSE150-LUC cells of human esophageal squamous cell carcinoma were resuspended in 100 µL PBS buffer and injected into the mice through the tail vein. After 5 to 6 weeks of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally for in vivo fluorescence photography. The mice with lung metastasis were dissected and the lungs with KYSE150-LUC cells were removed. The lungs were minced into minced meat and resuspended in culture medium. After standing for one week, the cells were washed twice with PSB buffer and Blasticidin (5 µg / mL, purchased from Shanghai Bio-Tech Biotechnology Co., Ltd.) was added. The cells were screened for one week until there was no cell death. These cells were KYSE150-LUC-LM1. Then, 0.25% trypsin was used to digest the LUCI-labeled KYSE150-LUC-LM1 cells into single cells. 10 6 Individual esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-LUC-LM1 were resuspended in 100 µL PBS buffer and injected into mice through the tail vein. After 5 to 6 weeks of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally and in vivo fluorescence photography was performed. Mice with lung metastasis were dissected and the lungs with KYSE150-LUC-LM1 cells were removed. The lungs were minced into minced meat and resuspended in culture medium. After standing for one week, the cells were washed twice with PSB buffer and Blasticidin (5 µg / mL, purchased from Shanghai Bio-Tech Biotechnology Co., Ltd.) was added. The cells were screened for one week until there was no cell death. These cells were KYSE150-LUC-LM2. After three cycles, KYSE150-LUC-LM3 cells were obtained.

[0042] KYSE410-Luc-I3 cell construction method: 0.25% trypsin was used to digest the LUCI-labeled KYSE410-Luc cells of human esophageal squamous cell carcinoma into single cells. The KYSE410-Luc cells were inoculated in the upper chamber membrane in a medium without fetal bovine serum (FBS) containing Matrigel (BD Biosciences), and a medium containing 10% FBS was added to the lower chamber. The cells were cultured at 37°C and 5% CO2 for 36 hours. The cells in the lower chamber were cells with invasive ability. The cells in the lower chamber were digested with 0.25% trypsin and then expanded and cultured again with 1640 medium. This was KYSE410-Luc-I1. After three cycles, KYSE410-Luc-I3 cells were obtained.

[0043] Example 1: lncRNA CTC-490G23.2 gene can enhance the metastasis and invasion ability of human esophageal squamous cell carcinoma cells

[0044] (1) Cell invasion assay

[0045] lncRNA CTC-490G23.2 stably overexpressing cells KYSE150-Luc-CTC-490G23.2 and KYSE410-Luc-CTC-490G23.2 and their control cells were seeded in the upper chamber membrane in a medium without fetal bovine serum (FBS) containing Matrigel (BD Biosciences), and 500 μL of medium containing 10% FBS was added to the lower chamber. Among them, 1) 500,000 KYSE150-Luc-CTC-490G23.2 and its control cells were plated in each chamber and cultured at 37°C, 5% CO2 for 36 hours; 2) 200,000 KYSE4100-Luc-CTC-490G23.2 and its control cells were plated in each chamber and cultured at 37°C, 5% CO2 for 24 hours. Finally, the chamber was washed twice with PBS buffer, and 500 μL of methanol was added to the lower chamber for fixation for 10 minutes. Remove the ethanol, add 0.2% crystal violet to the lower chamber for staining for 10 minutes, absorb the crystal violet and wash away the residual crystal violet with clean water, then dry it and observe it under an inverted microscope.

[0046] (2) Animal experiments

[0047] Twelve 6-week-old female mice (NCG) were randomly assigned to different groups (6 mice in each group).

[0048] 1) Human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-Luc and KYSE150-Luc-CTC-490G23.2 were digested into single cells using 0.25% trypsin and resuspended in PBS buffer.

[0049] 2) Anesthetize the mice and evaluate the degree of anesthesia through painless and painful stimulation to ensure that the mice are in an anesthetized state.

[0050] 3) The digested single cells were subcutaneously inoculated into the left hind foot pad of mice, 500,000 cells per mouse, for a total of 12 mice.

[0051] 4) After 6 to 7 weeks of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally, and in vivo fluorescence photography was performed to observe the lung metastasis of cancer cells.

[0052] 5) All excised footpad tumors and lymph nodes were fixed with 10% formaldehyde and subjected to histological analysis.

[0053] The results are as follows Figure 2 and Figure 3 As shown. Figure 2 The results of in vitro experiments showed that the CTC-490G23.2 gene can enhance the cell invasion ability of human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-Luc and human esophageal squamous cell carcinoma cells KYSE410-Luc. Figure 3 The in vivo experimental results show that the CTC-490G23.2 gene can promote the lymphatic metastasis of human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-Luc in mice.

[0054] Example 2: ASO screening targeting lncRNA CTC-490G23.2

[0055] (1) Invasion experiment

[0056] KYSE150-Luc-LM3 and KYSE410-Luc-I3 cells were transfected with 100 nM CTC-490G23.2-ASO#1, CTC-490G23.2-ASO#2, CTC-490G23.2-ASO#3, CTC-490G23.2-ASO#4 and their controls using Lipofectamine™ 3000 (purchased from Ththermo), and then cultured for 36 h at 37°C and 5% CO2. The cells were then seeded in the upper chamber membrane in FBS-free medium containing Matrigel (BD Biosciences), and 500 μL of medium containing 10% FBS was added to the lower chamber. 1) 500,000 KYSE150-Luc-LM3 and its control cells were plated in each chamber and cultured for 36 h at 37°C and 5% CO2. 2) KYSE410-Luc-I3 and its control cells were plated in each chamber with 200,000 cells and cultured at 37°C and 5% CO2 for 24 hours. Finally, the chamber was washed twice with PBS buffer, 500 μL of methanol was added to the lower chamber, and fixed for 10 minutes. After removing the ethanol, 0.2% crystal violet was added to the lower chamber for staining for 10 minutes. After absorbing the crystal violet, the residual crystal violet was washed off with water, and observed under an inverted microscope after drying. Among them, the four ASO sequences targeting lncRNA CTC-490G23.2 are:

[0057] CTC-490G23.2-ASO#1: 5'-GUCCATTACCTCTTTCCAGC-3' (full chain phosphorothioate modification + 2'OME modification at both ends)

[0058] CTC-490G23.2-ASO#2: 5'-GGAGAGGAGCTTAGCUCCAG-3' (full chain phosphorothioate modification + 2'OME modification at both ends)

[0059] CTC-490G23.2-ASO#3: 5'-UUCUGAGACAAGGCAAGUCC-3' (full chain phosphorothioate modification + 2'OME modification at both ends)

[0060] CTC-490G23.2-ASO#4: 5'-CUUCUTTTGTGATTCUUCAG-3' (full-chain phosphorothioate modification + 2'OME modification at both ends).

[0061] (2) qPCR experiment

[0062] RNA was extracted using TRIzol reagent, and cDNA was synthesized using PrimeScript RT reagent kit (Takara). lncRNA CTC-490G23.2 expression was measured by TransStart® Green qPCR SuperMix (TransGen Biotech, Beijing, China) and normalized to GAPDH.

[0063] The results are as follows Figure 4 and Figure 5 As shown. Figure 4 The in vitro experimental results show that after treatment with CTC-490G23.2-ASO#1 and CTC-490G23.2-ASO#2, the invasion of KYSE150-Luc-LM3 and KYSE410-Luc-I3 cells was most significantly inhibited. Figure 5 The results showed that CTC-490G23.2-ASO#2 reduced the expression of ncRNA CTC-490G23.2 most significantly. Therefore, CTC-490G23.2-ASO#2 was used in the subsequent experiments.

[0064] Example 3: Preparation of ASO / SV40-LAH4-L1 peptide nanomedicine targeting lncRNA CTC-490G23.2

[0065] SV40-LAH4-L1 or LAH4-L1 peptide (1 μg / μL) dissolved in nuclease-free water was mixed with ASO-CTC-490G23.2 (referred to as CTC-490G23.2-ASO#2) or ASO-CON (1 μg / μL) and vortexed for 1 min. The polydispersity index (PDI) and zeta potential of the nanoparticles after standing at room temperature for 30 min were measured using Nano ZS ZEN 3600 (Malvern Panalysis Ltd, UK). For morphological studies, a drop of the mixture was placed on a standard carbon-coated copper transmission electron microscopy (TEM) grid and then dried at room temperature for 6 h. The dried specimen was examined by TEM (JEOL Ltd., Tokyo, Japan). After confirmation of various indicators, the successfully prepared ASO / SV40-LAH4-L1 peptide nanodrug targeting lncRNA CTC-490G23.2 was experimentally verified. The results are shown in Figure 6 As shown, the PDI and zeta potential of Cy5-ASO-CTC-490G23.2 / SV40-LDH4-L1 nanodrug are within the range of normal nanoparticles.

[0066] Example 4: Characterization of ASO / SV40-LAH4-L1 peptide nanomedicine targeting lncRNA CTC-490G23.2

[0067] (1) Gel retardation test

[0068] Nanomedicine was prepared by SV40-LAH4-L1 and ASO-CTC-490G23.2 solutions at different ASO-CTC-490G23.2 / SV40-LAH4-L1 weight ratios of 5:1, 10:1, 15:1 and 20:1. The ASO-CTC-490G23.2 / SV40-LAH4-L1 complex was then loaded into agarose gel (2% w / v) and stained with ethidium bromide. Gel electrophoresis was performed in TAE buffer and observed under ultraviolet irradiation. The results are shown in Figure 2. Figure 7 As shown, the optimal ratio of SV40-LAH4-L1:ASO-CTC-490G23.2 is 20:1.

[0069] (2) Immunofluorescence experiment

[0070] The ASO used for immunofluorescence experiments was labeled with Cy5 fluorescence. Cy5-ASO-CTC-490G23.2 / SV40-LDH4-L1 or Cy5-ASO-CTC-490G23.2 / LDH4-L1 synthetic nanomedicines were added to KYSE150-Luc cells and cultured at 37°C and 5% CO2 for 24 h. The cells were then stained with Hoechst 33342 (Beyotime, Shanghai, China) (1.0 μg / mL) at 37°C for 20 min. Finally, a Zeiss LSM800 laser scanning confocal microscope (Carl Zeiss, Jena, Germany) was used for visualization. Images and colocalization analysis were obtained using ZEN2.3 pro software (Carl Zeiss). The results are shown in Figure 2. Figure 8 As shown, Cy5-ASO-CTC-490G23.2 / SV40-LDH4-L1 nanomedicine has the effect of targeting the cell nucleus.

[0071] Example 5: Functional verification of ASO / SV40-LAH4-L1 peptide nanodrug targeting lncRNA CTC-490G23.2

[0072] (1) Invasion experiment

[0073] ASO-CTC-490G23.2 / SV40-LDH4-L1 or Cy5-ASO-CTC-490G23.2 / LDH4-L1 synthetic nanomedicines prepared at the optimal ratio and their controls were added to KYSE150-Luc-LM3 and KYSE410-Luc-I3 cells at a concentration of 100 nM and cultured at 37°C, 5% CO2 for 36 h. The cells were then seeded in the upper chamber membrane in FBS-free medium containing Matrigel (BD Biosciences), and 500 μL of medium containing 10% FBS was added to the lower chamber. Among them, 1) KYSE150-Luc-LM3 and its control cells were plated at 500,000 per chamber and cultured at 37°C, 5% CO2 for 36 h. 2) KYSE410-Luc-I32 and its control cells were plated at 200,000 per chamber and cultured at 37°C, 5% CO2 for 24 h. Finally, the chamber was washed twice with PBS buffer, 500 μL methanol was added to the lower chamber and fixed for 10 min. The ethanol was removed, 0.2% crystal violet was added to the lower chamber for staining for 10 min, the crystal violet was sucked off and then washed with water to remove the residual crystal violet, and then observed under an inverted microscope after drying.

[0074] (2) Animal experiments

[0075] Forty-two 6-week-old female mice (NCG) were selected and randomly assigned to different groups, with a total of 7 groups, each with 6 mice.

[0076] 1) Human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-Luc-LM3 were digested into single cells using 0.25% trypsin and resuspended in PBS buffer.

[0077] 2) Anesthetize the mice and evaluate the degree of anesthesia through painless and painful stimulation to ensure that the mice are in an anesthetized state.

[0078] 3) The digested single cells were injected into the tail vein of mice, 1 million cells per mouse, for a total of 42 mice.

[0079] 4) After 15 days of feeding, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally, and in vivo fluorescence photography was performed to observe the lung metastasis of cancer cells. Nano drug treatment was performed after the start of lung metastasis was confirmed.

[0080] 5) Nanodrug treatment: The mice were treated with drugs by tail vein injection, once every 3 days, for four times, and the dosing schedule for each time was as follows:

[0081] Group 1: PBS

[0082] Group 2: ASO-CON / SV40-LDH4-L1, 0.5 mg / kg

[0083] Group 3: ASO-CTC-490G23.2 / SV40-LDH4-L1, 0.25 mg / kg

[0084] Group 4: ASO-CTC-490G23.2 / SV40-LDH4-L1, 0.5 mg / kg

[0085] Group 5: ASO-CON / LDH4-L1, 0.5 mg / kg

[0086] Group 6: ASO-CTC-490G23.2 / LDH4-L1, 0.25 mg / kg

[0087] Group 7: ASO-CTC-490G23.2 / LDH4-L1, 0.5 mg / kg;

[0088] After the treatment cycle, D-Luciferin (purchased from GOLDBIO) was injected intraperitoneally, and in vivo fluorescence photography was performed to observe the metastasis of cancer cells.

[0089] 6) Remove the liver, lungs, kidneys, etc. of the mice, fix them with 10% formaldehyde and perform histological analysis.

[0090] The results are as follows Fig. 9 and Fig.10 As shown. Fig. 9 The in vitro experimental results show that ASO-CTC-490G23.2 / SV40-LDH4-L1 nanomedicine treatment inhibited the invasion of KYSE150-Luc-LM3 and KYSE410-Luc-I3 cells. Fig.10 The in vivo experimental results show that 0.25mg / kg and 0.5mg / kg of ASO-CTC-490G23.2 / SV40-LDH4-L1 nanomedicine can reduce the bioluminescence in the lungs of mice. The results show that ASO-CTC-490G23.2 / SV40-LDH4-L1 nanomedicine can inhibit the metastasis of human esophageal squamous cell carcinoma luciferase-labeled cells KYSE150-Luc-LM3 in mice.

[0091] The liver, lung, kidney, heart and spleen of mice treated with nanomedicine were removed for immunohistochemistry. The results showed that (e.g. Fig.11 ), the experimentally synthesized nanomedicine had no effect on the liver, lung, kidney, heart, and spleen functions of mice.

[0092] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of an inhibitor targeting lncRNA CTC-490G23.2 in the preparation of an anti-esophageal cancer drug, characterized in that: The sequence of the lncRNA CTC-490G23.2 is shown in SEQ ID NO.1; The inhibitor is an antisense oligonucleotide composed of the following sequence: 5'-GGAGAGGAGCTTAGCUCCAG-3', with full-chain phosphorothioate modification and two-terminal 2'OME modification; The esophageal cancer is esophageal squamous cell carcinoma.

2. The use of the inhibitor targeting lncRNA CTC-490G23.2 according to claim 1 in the preparation of an anti-esophageal cancer drug, characterized in that: The inhibitor is an antisense oligonucleotide delivery nanoparticle constructed based on a cell-penetrating peptide, and the cell-penetrating peptide consists of the following sequence: PKKKRKV-GCG-KKALLAHALHLLALLALHLAHALKKA; The antisense oligonucleotide consists of the following sequence: 5'-GGAGAGGAGCTTAGCUCCAG-3', whole-chain phosphorothioate modification plus 2'OME modification at both ends; The esophageal cancer is esophageal squamous cell carcinoma.

3. Use of the inhibitor targeting lncRNA CTC-490G23.2 according to claim 2 in the preparation of an anti-esophageal cancer drug, characterized in that: The antisense oligonucleotide delivery nanoparticles are prepared by the following method: the cell penetrating peptide and the antisense oligonucleotide are mixed in nuclease-free water and vortexed to self-assemble and form the antisense oligonucleotide delivery nanoparticles.

4. Use of the inhibitor targeting lncRNA CTC-490G23.2 according to claim 3 in the preparation of an anti-esophageal cancer drug, characterized in that: The weight ratio of the cell penetrating peptide to the antisense oligonucleotide is 20:1; The effective dosage of the antisense oligonucleotide delivery nanoparticles is 0.25-0.5 mg / kg.

5. Use of an inhibitor targeting lncRNA CTC-490G23.2 according to any one of claims 1 to 4 in the preparation of an anti-esophageal cancer drug, characterized in that: The anti-esophageal cancer drug is a drug for inhibiting the invasion and metastasis of esophageal squamous cell carcinoma cells.

6. Use of an inhibitor targeting lncRNA CTC-490G23.2 according to any one of claims 1 to 4 in the preparation of an anti-esophageal cancer drug, characterized in that: The anti-esophageal cancer drug is a drug for treating and / or preventing esophageal squamous cell carcinoma.

7. An anti-esophageal cancer drug, characterized in that: An inhibitor targeting lncRNA CTC-490G23.2 in the use according to any one of claims 1 to 6.

8. The anti-esophageal cancer drug according to claim 7, characterized in that: The anti-esophageal cancer drug further comprises a pharmaceutically acceptable excipient; the pharmaceutically acceptable excipient is at least one of a sustained-release agent, a binder, a wetting agent, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant and a lubricant.

9. The anti-esophageal cancer drug according to claim 7, characterized in that: The anti-esophageal cancer drug is administered by tail vein injection.

Images