Cancer chemotherapy drug sensitization method, its sensitizer composition and use thereof

Abstract

A method for sensitizing a cancer chemotherapy drug includes: transfecting a cancer cell with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) to form a transgenic cancer cell, thereby blocking the subsequent effects of miR-1976; and applying a cancer composition to the transgenic cancer cell, at which point the transgenic cancer cell has been blocked from the subsequent effects of miR-1976, and because the transgenic cancer cell has been blocked from the subsequent effects of miR-1976, the transgenic cancer cell develops a sensitivity to the cancer composition.

Description

Technical Field

[0001] This invention relates to a method for sensitizing cancer chemotherapy drugs, a sensitizing agent composition (or testing reagent) thereof, and its use; particularly to a method for increasing the sensitization of cancer cells to chemotherapy drugs for cancer or pancreatic cancer, a sensitizing agent composition (or testing reagent) thereof, and its use; particularly to a method for sensitizing cancer (e.g., brain cancer, nasopharyngeal cancer, oral cancer, laryngeal cancer, esophageal cancer, gastric cancer, liver cancer, colorectal cancer, lung cancer, breast cancer, prostate cancer, bladder cancer, cervical cancer, germ cell cancer, skin cancer, osteosarcoma, or leukemia) or pancreatic cancer chemotherapy drugs that use miR-1976 to inhibit nucleic acids, a sensitizing agent composition (or testing reagent) thereof, and its use. Background Technology

[0002] Generally speaking, cancer includes brain cancer, nasopharyngeal cancer, oral cancer, laryngeal cancer, esophageal cancer, stomach cancer, liver cancer, colorectal cancer, pancreatic cancer, lung cancer, breast cancer, prostate cancer, bladder cancer, cervical cancer, germ cell cancer, skin cancer, osteosarcoma, and blood cancers (including lymphoma, leukemia, and multiple myeloma). For example, pancreatic cancer is one of the most common cancers in the world, and it is more common in developed countries. However, the prognosis of pancreatic adenocarcinoma is usually very poor, with a one-year survival rate of only 25% and a five-year survival rate of only 5%. Although the five-year survival rate increases to about 20% with early diagnosis, it is still not effectively controlled.

[0003] Regarding existing technologies related to microRNA miR-1976, US Patent No. US-8969315, "ENHANCEMENT OF PLACENTAL STEM CELL POTENCY USING MODULATORY RNA MOLECULES," only discloses miR-1976 itself, and has absolutely no relation to the technology of using miR-1976 to inhibit nucleic acids and the anti-apoptotic ability of cancer cells.

[0004] Another existing technology related to microribonucleic acid miR-1976, US Patent Publication No. US-20140080894, entitled "Enhanced Biodisturiction of OLGOMERS", only discloses miR-1976, and it is completely unrelated to the technology of using miR-1976 to inhibit nucleic acids and cancer cells' anti-apoptotic ability.

[0005] Another existing technology related to microRNA miR-1976, US Patent Publication No. US-20180237772, entitled "HYBRID tRNA / pre-miRNA MOLECULES AND METHODS OF USE", only discloses miR-1976 and has nothing to do with using miR-1976 to inhibit nucleic acid technology.

[0006] Another existing technology related to the gene XAF1, US Patent No. US-8021840, "DIAGNOSTIC MARKERFOR INTERFERON RESPONSIVENESS IN MULTIPLE SCLEROSIS", only discloses the gene XAF1, and has nothing to do with the technology of using miR-1976 to inhibit nucleic acids and the corresponding anti-apoptotic ability of cancer cells.

[0007] Another existing technology related to the gene XAF1, US Patent Publication No. US-20070218493, entitled "DIAGNOSTIC MARKER FOR INTERFERON RESPONSIVENESS IN MULTIPLE SCLEROSIS", only discloses the gene XAF1, and it has absolutely nothing to do with the technology of using miR-1976 to inhibit nucleic acids and the corresponding anti-apoptotic ability of cancer cells.

[0008] Another existing technology related to the gene XAF1, US Patent Publication No. US-20090215895, "THERAPEUTIC AND CARRIER MOLECULES", only discloses the gene XAF1, and it has nothing to do with the technology of using miR-1976 to inhibit nucleic acids and the corresponding anti-apoptotic ability of cancer cells.

[0009] Another existing technology related to the gene XAF1, US Patent Publication No. US-20170003277, entitled "BIOLOGICAL CHARATERIZATION OF A GLATIRAMER ACETATE RELATED DRUG PRODUCTUSING MAMMALIAN AND HUMAN CELLS", only discloses the gene XAF1, and it has absolutely nothing to do with the technology of using miR-1976 to inhibit nucleic acids and the corresponding anti-apoptotic ability of cancer cells.

[0010] Another existing technology related to the gene XAF1, US Patent Publication No. US-20180369303, "ONCOLYTICRHABDOVIRUS", only discloses the gene XAF1, and it has nothing to do with the technology of using miR-1976 to inhibit nucleic acids and the corresponding anti-apoptotic ability of cancer cells.

[0011] However, the application technologies of the microRNA miR-1976 or the related gene XAF1 in the aforementioned US patent applications No. US-8969315, US-20140080894, US-20180237772, US-8021840, US-20070218493, US-20090215895, US-20170003277, and US-20180369303 still require further improvement.

[0012] Obviously, the aforementioned U.S. Patent Nos. US-8969315, US-20140080894, US-20180237772, US-8021840, US-20070218493, US-20090215895, US-20170003277, and US-20180369303 are merely references and illustrations of the technical background and current state of technological development of this invention, and are not intended to limit the scope of this invention. Summary of the Invention

[0013] The main objective of this invention is to provide a method for sensitizing cancer chemotherapy drugs, a sensitizing agent composition (or diagnostic reagent) thereof, and its use. The method involves transfecting a cancer cell with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) to form a transfected cancer cell, thereby blocking the subsequent effects of miR-1976. A cancer composition is then applied to the transfected cancer cell, at which point the subsequent effects of miR-1976 are blocked. Because the subsequent effects of miR-1976 are blocked, the transfected cancer cell forms a cancer cell that has blocked the effects of miR-1976. Therefore, the transfected cancer cell develops sensitivity to the cancer composition, thereby achieving the effect of sensitizing cancer chemotherapy drugs or cancer diagnostic reagents.

[0014] To achieve the above objectives, a preferred embodiment of the cancer chemotherapy drug sensitization method of the present invention includes:

[0015] Transplanting a cancer cell with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) to form a transgenic cancer cell in order to block the subsequent effects of miR-1976.

[0016] A cancer composition is applied to the transgenic cancer cells, where the subsequent effects of miR-1976 have been blocked; and

[0017] Since the transgenic cancer cells have been blocked from the subsequent effects of miR-1976, thus forming a cancer cell that has blocked the effects of miR-1976, the transgenic cancer cells have developed a sensitivity to the cancer composition.

[0018] The preferred embodiment of the cancer chemotherapy drug sensitization method of the present invention further includes:

[0019] Search for at least one target molecule within the cancer cells;

[0020] Screening for at least one microRNA-binding target in a predetermined cell;

[0021] The target microRNA was replicated using a binding target nucleic acid fragment;

[0022] Sequence analysis was performed using the target nucleic acid fragment to determine a target gene and its corresponding position, thereby obtaining several miR-1976 binding targets; and

[0023] The target of miR-1976 binding was used to find a gene XAF1, and a cancer cell transfection antisense oligonucleotide program (e.g., cancer cell transfection siRNA-1976 program or cancer cell transfection antagomir-1976 program) was used to transfect the cancer cells so that the cancer cells could inhibit miR-1976.

[0024] In a preferred embodiment of the present invention, the microRNA binding target is screened using a specific gene selection method.

[0025] In a preferred embodiment of the present invention, the target gene and its corresponding location are determined by blast analysis.

[0026] The expression level of miR-1976 in the preferred embodiment of the present invention was obtained by microarray analysis or qPCR analysis.

[0027] To achieve the above objectives, the cancer chemotherapy drug sensitizing composition of a preferred embodiment of the present invention comprises:

[0028] A cancer cell is transfected with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976), which utilizes a cancer cell transfection procedure (e.g., a cancer cell transfection siRNA-1976 procedure or a cancer cell transfection antagomir-1976 procedure) to form a transfected cancer cell, so that the cancer cell can inhibit miR-1976; and

[0029] A cancer composition (e.g., an anticancer component) that is combined with the transfected cancer cells to form a composition that increases sensitivity;

[0030] The cancer composition is applied to the transgenic cancer cells, where the subsequent effects of miR-1976 are blocked, and the transgenic cancer cells form a cancer cell that has blocked the effects of miR-1976 because the subsequent effects of miR-1976 are blocked. Therefore, the transgenic cancer cells develop a sensitivity to the cancer composition.

[0031] The increased sensitivity composition of the preferred embodiment of the present invention results in a reduction of miR-1976 in an extracellular vesicle.

[0032] The increased sensitivity composition of the preferred embodiment of the present invention results in an increase in a cancer cell apoptosis marker.

[0033] In a preferred embodiment of the present invention, the cancer cell apoptosis marker is cleaved PARP.

[0034] In a preferred embodiment of the present invention, the cancer cell apoptosis marker corresponds to a gene XAF1.

[0035] To achieve the above objectives, a preferred embodiment of the pancreatic cancer chemotherapy drug sensitization method of the present invention includes:

[0036] Transplanting a pancreatic cancer cell with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) to form a transgenic pancreatic cancer cell in order to block the subsequent effects of miR-1976.

[0037] A pancreatic cancer composition was applied to the transgenic pancreatic cancer cells, where the subsequent effects of miR-1976 had been blocked; and

[0038] Since the transgenic pancreatic cancer cells have been blocked from the subsequent effects of miR-1976, thus forming pancreatic cancer cells that have blocked the effects of miR-1976, the transgenic pancreatic cancer cells develop a sensitivity to the pancreatic cancer composition.

[0039] The preferred embodiment of the pancreatic cancer chemotherapy drug sensitization method of the present invention further includes:

[0040] To search for at least one target molecule within these pancreatic cancer cells;

[0041] Screening for at least one microRNA-binding target in a predetermined cell;

[0042] The target microRNA was replicated using a binding target nucleic acid fragment;

[0043] Sequence analysis was performed using the target nucleic acid fragment to determine a target gene and its corresponding position, thereby obtaining several miR-1976 binding targets; and

[0044] The target of miR-1976 binding was used to find a gene XAF1, and a pancreatic cancer cell transfection antisense oligonucleotide program (e.g., pancreatic cancer cell transfection siRNA-1976 program or pancreatic cancer cell transfection antagomir-1976 program) was used to transfect the pancreatic cancer cells so that the pancreatic cancer cells could inhibit miR-1976.

[0045] In a preferred embodiment of the present invention, the microRNA binding target is screened using a specific gene selection method.

[0046] In a preferred embodiment of the present invention, the target gene and its corresponding location are determined by blast analysis.

[0047] The expression level of miR-1976 in the preferred embodiment of the present invention was obtained by microarray analysis or qPCR analysis.

[0048] To achieve the above objectives, the pancreatic cancer chemotherapy drug sensitizer composition of a preferred embodiment of the present invention comprises:

[0049] A pancreatic cancer cell transfection antisense oligonucleotide program (e.g., siRNA-1976 or antagomir-1976) utilizes a pancreatic cancer cell transfection antisense oligonucleotide program 2 (e.g., pancreatic cancer cell transfection siRNA-1976 program or pancreatic cancer cell transfection antagomir-1976 program) to form a transfected pancreatic cancer cell, which can then inhibit miR-1976; and

[0050] A pancreatic cancer composition (e.g., an anti-pancreatic cancer composition) combined with the transfected pancreatic cancer cells to form a composition that increases sensitivity;

[0051] When the pancreatic cancer composition is applied to the transgenic pancreatic cancer cells, the subsequent effects of miR-1976 are blocked, and pancreatic cancer cells that have blocked the effects of miR-1976 are formed. Therefore, the transgenic pancreatic cancer cells become sensitive to the pancreatic cancer composition.

[0052] The increased sensitivity composition of the preferred embodiment of the present invention results in a reduction of miR-1976 in an extracellular vesicle.

[0053] The increased sensitivity composition of the preferred embodiment of the present invention results in an increase in a pancreatic cancer cell apoptosis marker.

[0054] In a preferred embodiment of the present invention, the pancreatic cancer cells are labeled with cleaved PARP.

[0055] In a preferred embodiment of the present invention, the pancreatic cancer cell apoptosis marker corresponds to a gene XAF1.

[0056] The beneficial effects of this invention are as follows:

[0057] This invention provides a method for sensitizing cancer chemotherapy drugs, a sensitizing agent composition thereof, and its use, which can induce transgenic cancer cells to become sensitive to the cancer composition in order to achieve the effect of sensitizing cancer chemotherapy drugs or cancer testing reagents. Attached Figure Description

[0058] Figure 1 This is a flowchart illustrating a preferred embodiment of the cancer chemotherapy drug sensitization method of the present invention.

[0059] Figure 1A and Figure 1B This is a schematic diagram illustrating the expression levels of extracellular vesicles and intracellular microRNAs released by damaged cancer cells in a pancreatic cancer chemotherapy drug sensitization method according to a preferred embodiment of the present invention, using microarray analysis and qPCR analysis.

[0060] Figure 2A and Figure 2B The pancreatic cancer chemotherapy drug sensitization method of the preferred embodiment of the present invention uses specific gene selection technology to screen out intracellular miR-1976 binding targets, and then cross-references with bioinformatics data to show XAF1 as a potential target gene of miR-1976 related to cell apoptosis.

[0061] Figures 3A to 3C The diagram illustrates the pancreatic cancer chemotherapy drug sensitization method of the preferred embodiment of the present invention, which uses qPCR to analyze the expression of miR-1976 in cancer cells and the Western ink staining method to analyze the effect of the expression of miR-1976 in cancer cells on XAF1 protein expression and cell apoptosis.

[0062] Figure 4A and Figure 4B The preferred embodiment of the present invention describes the effect of transfection of antagomiR-1976 into pancreatic cancer cells on apoptosis using the Western ink staining method. The results are shown in schematic diagrams of the results obtained in pancreatic cancer BxPC-3 and AsPC-1 cells, respectively.

[0063] Figures 5A to 5D The preferred embodiment of the present invention describes the effect of transfecting siRNA-1976 into cancer cells on cell survival using a method for sensitizing pancreatic cancer chemotherapy drugs. The results are shown in the schematic diagrams for pancreatic cancer cells MIA Capa-2, BxPC-3, and AsPC-1. Detailed Implementation

[0064] To fully understand the present invention, preferred embodiments will be described in detail below with reference to the accompanying drawings, but these are not intended to limit the present invention.

[0065] The preferred embodiments of the present invention provide a method for sensitizing chemotherapy drugs for cancer, a sensitizing agent composition thereof, and its uses applicable to various cancers, including brain cancer, nasopharyngeal carcinoma, oral cancer, laryngeal cancer, esophageal cancer, gastric cancer, liver cancer, colorectal cancer, pancreatic cancer, lung cancer, breast cancer, prostate cancer, bladder cancer, cervical cancer, germ cell cancer, skin cancer, osteosarcoma, and blood cancers (including lymphoma, leukemia, and multiple myeloma). Another preferred embodiment of the present invention provides a method for sensitizing chemotherapy drugs for pancreatic cancer, a sensitizing agent composition (or testing reagent thereof), and its uses, providing a sensitizing agent for chemotherapy drugs to increase the sensitivity of pancreatic cancer cells to drugs, potentially achieving therapeutic effects at lower drug concentrations and thus increasing the efficacy of drug use.

[0066] The preferred embodiment of this invention describes a method for sensitizing pancreatic cancer chemotherapy drugs, its sensitizing agent composition (or diagnostic reagent), and its uses. These methods utilize the blocking of miR-1976 physiological effects to promote cancer cell sensitivity to drug treatment and inhibit cancer cell growth. Furthermore, microRNA molecules are used for screening molecular targets.

[0067] The preferred embodiment of the present invention employs a specific gene selection technique for the pancreatic cancer chemotherapy drug sensitization method, its sensitizing agent composition (or diagnostic reagent), and its uses. Furthermore, the preferred embodiment of the present invention uses the increase or decrease in the expression level of miR-1976 in a blood sample after chemotherapy drug injection as a diagnostic reagent.

[0068] Figure 1 A schematic flowchart illustrating a preferred embodiment of the cancer chemotherapy drug sensitization method of the present invention is provided. Please refer to... Figure 1As shown, for example, methods for sensitizing cancer chemotherapy drugs include:

[0069] A cancer cell 1 (or pancreatic cancer cell or other types of cancer cells) is selected to be transfected with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) using a cancer cell transfection antisense oligonucleotide procedure 2 (e.g., pancreatic cancer cell transfection siRNA-1976 procedure or pancreatic cancer cell transfection antagomir-1976 procedure) to form a transfected cancer cell 10 in order to block the subsequent effects of miR-1976.

[0070] A cancer composition 3 (e.g., an anticancer composition or other cancer chemotherapy composition) is appropriately applied to the transgenic cancer cells 10, wherein the transgenic cancer cells 10 have had their subsequent effects of miR-1976 blocked; and

[0071] Since the transgenic cancer cell 10 has been blocked from the subsequent effects of miR-1976 to form a cancer cell 11 that has blocked the effects of miR-1976, the transgenic cancer cell 10 is obviously sensitive to the cancer composition 3.

[0072] Please refer to again Figure 1 As shown, for example, the preferred embodiment of the cancer chemotherapy drug sensitization method of the present invention further includes:

[0073] Search for at least one target molecule within the cancer cell 1.

[0074] At least one microRNA-binding target is appropriately screened within a predetermined cell.

[0075] The target microRNA was replicated by appropriately binding a target nucleic acid fragment;

[0076] By appropriately analyzing the sequence of the target nucleic acid fragment and determining a target gene and its corresponding position, several miR-1976 binding targets can be obtained; and

[0077] Using these miR-1976 binding targets, a gene XAF1 was appropriately identified, and the pancreatic cancer cell transfection antisense oligonucleotide program 2 (e.g., pancreatic cancer cell transfection siRNA-1976 program or pancreatic cancer cell transfection antagomir-1976 program) was used to appropriately transfect the cancer cells so that the cancer cells 1 could appropriately inhibit miR-1976.

[0078] Please refer to again Figure 1As shown, for example, the pancreatic cancer cell transfection antisense oligonucleotide program 2 of the preferred embodiment of the present invention includes a pancreatic cancer cell transfection siRNA-1976 program or a pancreatic cancer cell transfection antagomir-1976 program.

[0079] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of the preferred embodiment of the present invention may be selected from a primary anti-metabolite drug (e.g., Methotrexate, Pemetrexed, Cytarabine, 5-Fluorouracil, Capecitabine, Gemcitabine, 6-Mercaptopurine, Azathioprine, Fludarabine, Cladribine, or Hydroxyurea).

[0080] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of another preferred embodiment of the present invention may also be selected from an alkylating agent (e.g., Cyclophosphamide, Ifosfamide, Chlorambucil, Melphalan, Temozolomide, Carmustine, Lomustine, Streptozocin, Busulfan, Procarbazine, Cisplatin, Carboplatin, or Oxaliplatin).

[0081] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of another preferred embodiment of the present invention may also be selected from a topoisomerase inhibitor (e.g., Irinotecan, Topotecan, Etoposide, or Doxorubicin).

[0082] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of another preferred embodiment of the present invention may also be selected from a mitotic inhibitor (e.g., Vincrisine, Vinblastine, Vinorelbine, Docetaxel, Paclitaxel, Eribulin, Ixabepilone, or Epothilone).

[0083] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of another preferred embodiment of the present invention may also be selected from an antitumor antibiotic (e.g., Bleomycin, Actinomycin D, doxorubicin, daunorubicin, idarubicin or Mitomycin).

[0084] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of another preferred embodiment of the present invention may also be selected from a tyrosine kinase inhibitor (e.g., Imatinib, Dasatinib, Nilotinib, Erlotinib, Gefitinib, Afatinib, Osimertinib, Alectinib, Crizotinib, Dabrafenib, Encorafenib, Vemurafenib, Trametinib, Ibrutinib, Ruxolitinib, or Palbociclib).

[0085] Please refer to again Figure 1 As shown, for example, the cancer composition 3 of another preferred embodiment of the present invention may also be selected from an anti-tumor and anti-tumor receptor antibody or other compositions (e.g., L-asparaginase, Bortezomib, Carfilzomib, Ixazomib or Olaparib).

[0086] Figure 1A and Figure 1B This diagram illustrates a preferred embodiment of the pancreatic cancer chemotherapy drug sensitization method, employing microarray analysis and qPCR to analyze the expression levels of extracellular vesicles and intracellular microRNAs released from damaged cancer cells. Please refer to... Figure 1A and Figure 1B As shown, for example, the levels of miR-1976, miR-877-3p, and miR-4728-3p increased the most, which can be confirmed by qPCR, with the increase in miR-1976 being the most significant.

[0087] Please refer to Figure 1B As shown, for example, the level of miR-1976 in pancreatic cancer cells can also be induced to increase by cellular damage caused by gemcitabine.

[0088] Figure 2A and Figure 2B The preferred embodiment of the present invention discloses a method for sensitizing pancreatic cancer chemotherapy drugs by employing specific gene selection technology to screen for intracellular miR-1976 binding targets, and then cross-referencing with bioinformatics data to obtain a schematic diagram showing XAF1 as a potential target gene of miR-1976 associated with apoptosis. Please refer to... Figure 2A and Figure 2BAs illustrated, in a preferred embodiment of the present invention, specific gene selection technology is used to screen for microRNA binding targets in specific cells. After replicating the target nucleic acid fragment, sequence analysis is performed, and BLAST analysis can be used to determine the target gene and its corresponding location. In a preferred embodiment of the present invention, several miR-1976 binding targets are obtained, and then cross-referenced with genes related to cell death. Using the miR-1976 target group predicted by the targetScan tool, the gene XAF1, which is related to apoptosis, is obtained, and thus becomes the regulatory target of miR-1976.

[0089] Figures 3A to 3C This diagram illustrates a preferred embodiment of the present invention, showing the selection of a pancreatic cancer chemotherapy drug sensitization method using the Western ink staining method to analyze the effect of a large amount of miR-1976 expressed in cancer cells on cell apoptosis. Please refer to... Figures 3A to 3C As illustrated, for example, a preferred embodiment of the present invention expresses a large amount of miR-1976 in cancer cells to simulate the increase in miR-1976 when cells are damaged, or the effect of cells being exposed to extracellular vesicles containing miR-1976. When cells express a large amount of miR-1976, in addition to a relative decrease in XAF1 levels, the apoptosis marker cleaved PARP in pancreatic cancer cells is also significantly reduced.

[0090] Figure 4A and Figure 4B This diagram illustrates the effect of Western ink staining analysis on apoptosis in pancreatic cancer cells after transfection with antagomiR-1976, revealing a preferred embodiment of the present invention. Please refer to... Figure 4A As shown, for example, are results obtained using a preferred embodiment of the invention in pancreatic cancer BxPC-3 cells. Please refer to... Figure 4B As shown, for example, the results of a preferred embodiment of the present invention in pancreatic cancer AsPC-1 cells.

[0091] Please refer to Figure 4A and Figure 4B As shown, for example, in a preferred embodiment of the present invention, pancreatic cancer cells are first transfected with antagomir-1976 to block the subsequent effects of miR-1976, thus significantly increasing the apoptosis marker cleaved PARP.

[0092] For example, in the preferred embodiments of the present invention, the method for sensitizing cancer chemotherapy drugs, the sensitizing agent composition thereof, and its use can be primarily selected to replace "antagomir-1976" with "inhibit nucleic acid-1976".

[0093] For example, the preferred embodiment of the present invention may employ antagomir, which has a special structure, and the combination of the special structure forms an interfering nucleic acid molecule of antagomir, and antagomir has RNA sequence specificity, namely the specific sequence “ACAGCAA” (Seq ID No. 1).

[0094] For example, in a preferred embodiment of the present invention, the 20 nucleic acid sequences of Antagomir-1976 may be selected as follows:

[0095] Seq ID No. 2: 5'-ACAGCAAGGAGGGCAGGAGG-3'.

[0096] For example, another preferred embodiment of the present invention may use Antagomir-1976 having a key region for inhibiting nucleic acid-1976, and the key region contains a specific sequence of “ACAGCAA” and its nucleic acid length is 17 to 42 bases.

[0097] For example, another preferred embodiment of the present invention may use Antagomir-1976 with the sequence 5'-(N)xACAGCAA(N)y, which inhibits nucleic acid-1976; where N is A, U, G, or C; and x = 0 to 5; y = 10 to 30. It can also be represented as Seq ID No. 3: 5'-NACAGCAAN-3'; where N is A, U, G, or C; and the number of Ns at the 5' end is 0 to 5; the number of Ns at the 3' end is 10 to 30.

[0098] Figures 5A to 5D The preferred embodiment of the present invention discloses the effect of transfecting siRNA-1976 into pancreatic cancer cells on cell survival using cell survival analysis. The schematic diagrams show the results obtained from transfecting siRNA-1976 into pancreatic cancer cells MIA Capa-2, BxPC-3, and AsPC-1. Please refer to... Figures 5A to 5D As shown, for example, the preferred embodiment of the present invention requires the use of the sensitizer siRNA-1976 (or a composition with similar sensitizing function) and the chemotherapy drug in order to affect the survival rate of the cancer cells.

[0099] Please refer to again Figures 5A to 5D As shown, for example, if only the sensitizer siRNA-1976 (or a composition with similar sensitizing function) is used without chemotherapy drugs, the use of the sensitizer siRNA-1976 (or a composition with similar sensitizing function) will not affect the survival rate of cancer cells.

[0100] The above experimental data are preliminary results obtained under specific conditions and are only used to facilitate understanding or reference of the technical content of this invention. Further related experiments are required. This experimental data and its results are not intended to limit the scope of this invention.

[0101] The foregoing preferred embodiments are merely illustrative of the present invention and its technical features. The technology of these embodiments can still be implemented by various substantially equivalent modifications and / or substitutions. Therefore, the scope of the present invention shall be determined by the scope defined in the claims. SEQUENCE LISTING <110> Fu Jen Catholic University Foundation <120> Methods for sensitizing cancer chemotherapy drugs, their sensitizing agent compositions and their uses <160> 3 <170> PatentIn version 3.3 <210> 1 <211> 7 <212> RNA <213> Artificial sequence <220> <223> antagomir <400> 1 acagcaa 7 <210> 2 <211> 20 <212> RNA <213> Artificial sequence <220> <223> antagomir-1976 <400> 2 acagcaagga gggcaggagg 20 <210> 3 <211> 9 <212> RNA <213> Artificial sequence <220> <223> antagomir-1976 <220> <221> misc_feature <222> (1)..(1) <223> n = a, u, g, or c, and the number of n is between 0 and 5. <220> <221> misc_feature <222> (9)..(9) <223> n = a, u, g, or c, and the number of n is between 10 and 30. <400> 3 nacagcaan 9

Claims

1. The use of a sensitizing composition for a cancer chemotherapy drug in the preparation of a drug for treating pancreatic cancer, characterized in that, The sensitizing composition of this cancer chemotherapy drug contains a cancer cell transfection antisense oligonucleotide and a pancreatic cancer chemotherapy drug. Include: A pancreatic cancer cell was subjected to a transgenic antisense oligonucleotide procedure to form a transgenic pancreatic cancer cell in order to block the subsequent effects of miR-1976. A pancreatic cancer chemotherapy drug was administered to transfected pancreatic cancer cells following the antisense oligonucleotide transfection procedure, at which point the subsequent effects of miR-1976 had been blocked in the transfected pancreatic cancer cells; and Since the transgenic pancreatic cancer cells have been blocked from the subsequent effects of miR-1976 to form pancreatic cancer cells that have blocked the effects of miR-1976, the transgenic pancreatic cancer cells have developed sensitivity to the pancreatic cancer chemotherapy drugs. They achieve the sensitizing effect of pancreatic cancer chemotherapy drugs by using the transgenic antisense oligonucleotide program. The procedure for transfecting antisense oligonucleotides is the antagomir-1976 transfectation procedure, the sequence of which is shown in Seq ID No.

2.

2. The use of the sensitizing composition of the cancer chemotherapy drug as described in claim 1 in the preparation of a drug for treating pancreatic cancer, characterized in that, Also includes: Screening for at least one microRNA binding target in pancreatic cancer cells to obtain a miR-1976 binding target, the gene of which is [missing information]. XAF1 .

3. The use of the sensitizing composition of the cancer chemotherapy drug as described in claim 2 in the preparation of a drug for treating pancreatic cancer, characterized in that, The target microRNA binding agent was screened using a specific gene selection method.

4. The use of the sensitizing composition of the cancer chemotherapy drug as described in claim 2 in the preparation of a drug for treating pancreatic cancer, characterized in that, The target gene and its corresponding location were determined by blast analysis.

5. The use of the sensitizing composition of the cancer chemotherapy drug as described in claim 2 in the preparation of a drug for treating pancreatic cancer, characterized in that, The expression level of miR-1976 was obtained by microarray analysis or qPCR analysis.

6. A pancreatic cancer chemotherapy drug sensitizing composition, characterized in that, It consists of the following composition: A pancreatic cancer cell was transfected with antagomir-1976. The procedure involved transfecting a pancreatic cancer cell with antagomir-1976 to create a transfected pancreatic cancer cell, which could then inhibit miR-1976. A chemotherapy drug for pancreatic cancer, which is applied to pancreatic cancer cells that have been transfected with the antagomir-1976 procedure to form a composition that increases sensitivity. The pancreatic cancer chemotherapy drug is applied to pancreatic cancer cells that have been transfected with the antagomir-1976 program. At this time, the transfected pancreatic cancer cells have blocked the subsequent effects of miR-1976, and because the transfected pancreatic cancer cells have blocked the subsequent effects of miR-1976, they form pancreatic cancer cells that have blocked the effects of miR-1976. Therefore, the transfected pancreatic cancer cells have a sensitivity to the pancreatic cancer chemotherapy drug. The sequence of antagomir-1976 is shown in Seq ID No.2, and the chemotherapy drug for pancreatic cancer is gemcitabine.

7. The pancreatic cancer chemotherapy drug sensitizer composition as described in claim 6, characterized in that, This increased sensitivity composition resulted in a reduction of miR-1976 in an extracellular vesicle.

8. The pancreatic cancer chemotherapy drug sensitizer composition as described in claim 7, characterized in that, This sensitivity-enhancing composition leads to an increase in a cancer cell apoptosis marker.

9. The pancreatic cancer chemotherapy drug sensitizer composition as described in claim 8, characterized in that, The apoptosis marker for these cancer cells was cleaved PARP.

10. The pancreatic cancer chemotherapy drug sensitizer composition as described in claim 8, characterized in that, This cancer cell apoptosis marker corresponds to a gene. XAF1 .

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