KR101697771B1 - Anti-tumor composition comprising ITM2A polypeptide or DNA coding ITM2A gene - Google Patents

Abstract

The present invention relates to an anticancer composition comprising an ITM2A polypeptide as an active ingredient. More specifically, the present invention relates to an anticancer composition comprising a nucleic acid encoding an ITM2A polypeptide as an active ingredient. The ITM2A polypeptide of the present invention can be used as an effective ingredient to inhibit the child's action and further effectively kill cancer cells and the like. The composition of the present invention has an effect of inhibiting the action of the child to make the cancer cells more responsive to the anticancer agent, thereby effectively killing the cancer cells.

Description

An anticancer composition comprising an ITM2A polypeptide or a nucleic acid encoding the ITM2A polypeptide as an active ingredient (Anti-tumor composition comprising ITM2A polypeptide or DNA coding ITM2A gene)

The present invention relates to an anticancer composition comprising an ITM2A polypeptide as an active ingredient. More specifically, the present invention relates to an anticancer composition comprising a nucleic acid encoding an ITM2A polypeptide as an active ingredient. The ITM2A polypeptide of the present invention can be used as an effective ingredient to inhibit the child's action and further effectively kill cancer cells and the like.

ITM2A stands for integral membrane protein 2A and is encoded by the ITM2A gene. The ITM2A protein performs functions related to differentiation of bone cells and chondrocytes. It also activates T cells and the immune system and is known to be involved in the differentiation of muscle cells.

Autophagy is a mechanism that breaks down mitochondria in cells. When a cell is deficient in energy, it becomes viable by obtaining energy through the action of degrading the organelles in the cell. Child action is used when there is insufficient energy, when bacteria are infected in the cytoplasm, or when organelles such as mitochondria are destroyed by reactive oxygen species (ROS) and can not function. The process of child action is surrounded by a membrane that separates the mitochondria (Isolation Membrane), then converted into autophagosome. It is then degraded by autoplysomes, which process is called autophagic flux (see FIG. 1). It has been known that childhood action is very important for the survival of cancer cells in recent years, and it is known that cancer cells are killed if the child action is excessively promoted or inhibited. In addition, recent studies have shown that inhibition of autophagosome accumulation in cells results in the death of cells or the death of cells, depending on the results.

Cancer is very difficult to treat now because it occurs in various forms by various causes. Therefore, various treatment methods should be developed. Cancer cells are reported to be sensitive to various anticancer drugs by suppressing the child action through various papers, and substances suppressing this child action can be useful for increasing the activity of anticancer drugs. Currently, there are inhibitory substances such as Chloroquine and Bafilomycin A1. However, there are no known inhibitors of gene action.

On the other hand, it is known that the ITM2A gene is associated with bone cell and chondrocyte differentiation, but it is not known whether the gene has anticancer activity.

Accordingly, the present inventors have completed the present invention by developing an anticancer composition comprising ITM2A polypeptide as an active ingredient.

Accordingly, an object of the present invention is to provide an anticancer composition comprising an ITM2A polypeptide as an active ingredient.

It is still another object of the present invention to provide an anticancer composition comprising a nucleic acid encoding an ITM2A polypeptide as an active ingredient.

In order to achieve the above object, the present invention provides an anticancer composition comprising ITM2A polypeptide as an active ingredient.

In order to achieve still another object of the present invention, there is provided an anticancer composition comprising a nucleic acid encoding ITM2 polypeptide as an active ingredient.

Hereinafter, the present invention will be described in detail.

Unless defined otherwise, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains and are consistent with what is described in the following references: Singleton et al. , J. Wiley & Sons, 2nd Ed, New York, 1994; and Janeway, C., Travers, P., Walport, M., Shlomchik, Garland Publishing, 5th Ed, New York, 2001].

The present invention provides an anticancer composition comprising an ITM2A polypeptide as an active ingredient. The ITM2A polypeptide is an integral membrane protein 2A and is encoded in the ITM2A gene. Proteins encoded by this gene participate in differentiation of bone cells and chondrocytes. It is also involved in T-cell activation of the immune system in muscle cell differentiation. Preferably, the ITM2A polypeptide of the invention is derived from Genbank Accession no. NP001165052.1 (isoform1) or NP004858.1 (isoform2), more preferably the amino acid sequence of SEQ ID NO: 1.

The "polypeptide" of the present invention includes not only chemically synthesized polypeptides but also naturally synthesized polypeptides encoded by the genes of the cultured cells and recombinant polypeptides secreted by the cells. A recombinant polypeptide is a polypeptide encoded by a transgene introduced into a cell by molecular biology techniques. Polypeptides may be modified by chemical methods or by enzymes in post-translational processing.

The composition of the present invention is characterized by inhibiting the cell function and promoting cell death.

The term " child action " of the present invention means a mechanism of decomposing mitochondria and the like in cells. When a cell is deficient in energy, it becomes viable by obtaining energy through the action of degrading the organelles in the cell. Child action is used when there is insufficient energy, when bacteria are infected in the cytoplasm, or when organelles such as mitochondria are destroyed by reactive oxygen species (ROS) and can not function. The process of child action is surrounded by a membrane that separates the mitochondria (Isolation Membrane), then converted into autophagosome. It is then broken down by the autoplysome, which is called the autophagic flux. It has been known that childhood action is very important for the survival of cancer cells in recent years, and it is known that cancer cells are killed if the child action is excessively promoted or inhibited. In addition, recent studies have shown that inhibition of autophagosome accumulation in cells results in the death of cells or the death of cells, depending on the results.

The " cancer " of the present invention generally represents or describes the physiological condition of a mammal having the characteristics of uncontrolled cell growth / proliferation.

The cancer of the present invention can be used for the treatment of prostate cancer, breast cancer, lung cancer, pancreatic cancer, glioma, ovarian cancer, liver cancer, cervical cancer, endometrial cancer, brain cancer, leukemia, colon cancer, glioblastoma, bladder cancer, skin cancer, Cervical cancer, oral cancer, stomach cancer, renal cancer, and bone cancer.

The anticancer agents of the present invention may be used in combination with fluorouracil, everlimus, metformin, doxorubicin, Epigallocatechin gallete (EGCG), trastuzumab, Rapamycin, cisplatin, and the like.

The pharmaceutical composition may be formulated in a suitable form together with a pharmaceutically acceptable carrier. &Quot; Pharmaceutically acceptable " refers to compositions which are physiologically acceptable and which, when administered to humans, do not normally cause allergic reactions such as gastrointestinal disorders, dizziness, or the like. Pharmaceutically acceptable carriers include, for example, carriers for oral administration such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like, water for parenteral administration such as water, suitable oils, saline, aqueous glucose and glycols And may further contain stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. Other pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, Pa., 1995).

The pharmaceutical composition of the present invention is not limited thereto, but may be formulated for oral administration. The nucleic acid and the pharmaceutically acceptable salt may be mixed with an excipient to prepare ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups and wafers , ≪ / RTI > These formulations may contain, in addition to the active ingredient, a diluent such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine and a lubricant such as silica, talc, stearic acid and its magnesium or calcium salt and / Or polyethylene glycol). The tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally mixed with starch, agar, A disintegrating agent such as sodium salt, an absorbent, a coloring agent, a flavoring agent and / or a sweetening agent. The formulations may be prepared by conventional mixing, granulating or coating methods. The injectable formulations may be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component may be formulated for injection by dissolving in saline or buffer. In the case of a preparation for parenteral administration, the composition can be formulated in the form of injections, creams, lotions, ointments, ointments, moisturizers, gels, aerosols and nasal inhalers by methods known in the art. These formulations are described in Remington's Pharmaceutical Science, 15th edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a commonly known formulary for all pharmaceutical chemistries.

The pharmaceutical composition formulated as described above may be administered in an effective amount through various routes including oral, transdermal, subcutaneous, intravenous, or muscular. The term "effective amount" as used herein refers to an amount that shows a therapeutic effect of cancer when administered to a patient. The "subject" may be an animal, preferably a mammal, particularly an animal including a human. Derived cells, tissues, organs, and the like. The subject may be a patient requiring treatment.

The pharmaceutical composition formulated as described above may be administered in an effective amount through various routes including oral, transdermal, subcutaneous, intravenous, or muscular. The term "effective amount" as used herein refers to an amount that shows a therapeutic effect of cancer when administered to a patient. The "subject" may be an animal, preferably a mammal, particularly an animal including a human. Derived cells, tissues, organs, and the like. The subject may be a patient requiring treatment.

The pharmaceutical composition of the present invention may be administered as it is or may be prepared and administered in various formulations as described above, preferably administered until the desired effect, that is, endothelial cell death and / or anti-cancer effect . The pharmaceutical composition of the present invention can be administered by various routes according to methods known in the art. In other words, it can be administered orally or parenterally such as oral, intramuscular, intravenous, intradermal, intraarterial, intramedullary, intrathecal, intraperitoneal, intranasal, vaginal, rectal, sublingual or subcutaneous, gastrointestinal, ≪ / RTI > For example, the pharmaceutical composition of the present invention may be directly applied to the skin, or the polypeptide may be prepared into a form for injection, and a predetermined amount may be injected into the lower layer of the skin with a 30 gauge thin injection needle, prick < / RTI > Preferably directly on the skin. In addition, the pharmaceutical composition of the present invention may be administered in the form of being bound to a molecule that causes high affinity binding to a target cell or tissue (for example, skin cells or dermal tissue), or encapsulated in the molecule. The pharmaceutical composition of the present invention may be administered orally or parenterally, e.g., in the form of a sterol (e.g., cholesterol), a lipid (e.g., a cationic lipid, a virosome or a liposome), or a target cell- specific binding agent Lt; / RTI > ligand). Suitable coupling or crosslinking agents may include, for example, protein A, carbodiimide, N-succinimidyl-3- (2-pyridyldithio) propiotate (SPDP), and the like.

The anticancer composition of the present invention is administered in combination with an anticancer agent.

The anticancer composition of the present invention is characterized in that it is administered simultaneously, separately, or sequentially with an anticancer agent.

In the pharmaceutical composition of the present invention, the total effective amount of the polypeptide of the present invention may be administered to a subject in a single dose, and the fractionated treatment in which multiple doses are administered for a prolonged period of time < / RTI > protocol). The pharmaceutical composition of the present invention may be administered several times a day at an effective dose of 0.1 μg to 1 g per one dose, although the content of the active ingredient may be varied depending on the purpose of administration. However, the effective dose is determined not only by the route of administration and the number of treatments but also by various factors such as the age, weight, health condition, sex, severity of disease, diet and excretion rate of the individual requiring treatment, Therefore, in view of this point, one of ordinary skill in the art will be able to determine an appropriate effective dose depending on the purpose of administration. The pharmaceutical composition of the present invention is not particularly limited to the formulation, administration route and administration method as long as the effect of the present invention is exhibited.

There is provided an anticancer composition comprising an ITM2A gene and a nucleic acid encoding a polypeptide as an active ingredient.

The nucleic acid of the present invention includes DNA or RNA. Preferably a DNA coding for the ITM2A gene or fragment thereof derived from a mammal, more preferably a human.

The nucleic acid may be introduced into an expression vector such as a plasmid or a viral vector by a known method, and then the expression vector may be introduced into the target cell in an expression form by infection or transduction by various methods known in the art Can be introduced.

The nucleic acid of the present invention is characterized by having the nucleotide sequence shown in SEQ ID NO: 2.

The nucleic acid of the present invention is characterized by being contained in an expression vector.

The above expression vector is a method of delivering plasmid DNA directly to human cells using an approved FDA-approved gene transfer method that can be used in humans (Nabel, EG, et al., Science, 249: 1285-1288, 1990). Unlike the viral vector, the expression vector DNA can be purified homogeneously. As the plasmid expression vector that can be used in the present invention, mammalian expression plasmids known in the art can be used. Examples include, but are not limited to pRK5 (European Patent No. 307,247), pSV16B (International Patent Publication No. 91/08291) and pVL1392 (PharMingen).

A plasmid expression vector containing the nucleic acid according to the present invention may be prepared by a method known in the art such as, but not limited to, transient transfection, microinjection, transduction, Cell fusion, calcium phosphate precipitation, liposome-mediated transfection, DEAE dextran-mediated transfection, polybrene-mediated transfection, , Electroporation, gene gun, and other known methods for introducing DNA into cells (Wu et al., J. Bio. Chem., 267 : 963-967, 1992; Wu and Wu, J. Bio. Chem., 263: 14621-14624, 1988).

The expression vector of the present invention is characterized by being a plasmid or a viral vector.

The viral vector of the present invention is characterized in that it is selected from retrovirus vectors, adenovirus vectors and herpes virus vectors.

In addition, virus expression vectors containing the nucleic acid according to the present invention include, but are not limited to, retrovirus, adenovirus, herpes virus, and avipoxvirus . The retroviral vector is constructed such that all of the viral genes have been removed or changed so that the non-viral proteins are made in the cells infected with the viral vector. The main advantage of retroviral vectors for gene therapy is that they transfer large numbers of genes into the cloned cells, accurately integrate the genes transferred into the cellular DNA, and do not lead to subsequent infection after gene transfection (Miller, AD, Nature , 1992,357: 455-460). FDA-approved retroviral vectors were prepared using PA317 amphotropic retroviral packaging cells (Miller, A.D. and Buttimore, C., Molec. Cell Biol., 6: 2895-2902, 1986). Non-retroviral vectors include the adenoviruses mentioned above (Rosenfeld, MA, et al., Cell, 68: 143-155, 1992; Jaffe, HA et al., Nature Genetics, 1: 372- 378, 1992; Lemarchand, P. et al., Proc. Natl. Acad. Sci. USA, 89: 6482-6486, 1992). The main advantage of adenoviruses is the ability to carry large quantities of DNA fragments (36 kb genome) and to infect very high reverse non-cloned cells. Herpesviruses may also be useful for human gene therapy (Wolfe, J. H., et al., Nature Genetics, 1: 379-384, 1992). In addition, a known appropriate viral vector may be used in the anticancer composition of the present invention.

The vector capable of expressing the ITM2A gene or a fragment thereof may be administered by a known method. For example, topically, parenterally, orally, nasally, intravenously, intramuscularly, subcutaneously, or by any other suitable means. In particular, the vector may be injected directly into a target cancer cell in an effective amount for treating cancer cells of the target tissue. In particular, in the case of cancer in the body cavity such as the eye, gastrointestinal tract, genitourinary tract, lung, and bronchial system, the anticancer composition of the present invention is injected into a hollow organ affected by the cancer, can be injected directly using a catheter or other type of transport tube. Imaging devices such as X-rays, sonograms, or fiberoptic visualization systems can then be used to locate the target tissue and inject the needle or catheter. In addition, in the case of cancer which can not be directly reached or analytically separated, the anticancer composition according to the present invention can be administered into the blood circulation system.

The present invention relates to an anticancer composition comprising an ITM2A polypeptide as an active ingredient. More specifically, the present invention relates to an anticancer composition comprising a nucleic acid encoding an ITM2A polypeptide as an active ingredient. The ITM2A polypeptide of the present invention can be used as an effective ingredient to inhibit the action of allergic agents and effectively kill cancer cells and the like. The composition of the present invention has an effect of inhibiting the action of the child to make the cancer cells more responsive to the anticancer agent, thereby effectively killing the cancer cells.

Fig. 1 is a schematic view showing the cell action of a cell.
FIG. 2 is a photograph showing the formation of a child action by ITM2A (left photograph: starved cell, middle photograph: cell treated with Bafilomycin A1, right photograph: cell expressed with ITM2A).
Fig. 3 is a photograph showing the inhibition of the subcultural flow by ITM2A. Fig.
FIG. 4 is a photograph showing that overexpression of ITM2A in cervical cancer cell line HeLa cell inhibits colony formation (upper: control, lower: test group overexpressing ITM2A gene).
FIG. 5 is a graph showing a decrease in the expression level of the ITM2A gene in ovarian cancer tissues (green bar: normal tissue, red bar: ovarian cancer tissue, 1-4: borderline tumor, 5-10: invasive tumor).

Hereinafter, the present invention will be described in detail.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

<Experimental Method>

Experimental cell culture and preparation of experimental material

HEK293 and HeLa cells were cultured in DMEM medium (Welgene, LM001-05) containing 10% fetal bovine serum (Gibco, 26140-079).

The HEK293 cell line expressing GFP-LC3 was obtained by transforming HEK293 cells with a plasmid encoding RFP-GFP-LC3. The RFP-GFP-LC3 plasmid used was obtained from T. Yoshimori. Thereafter, the transformed cells stably expressing RFP-GFP-LC3 were kept in a medium containing G418 (1 mg / ml) for 5 days.

For reporter analysis, HEK293 cells were transformed with lipofectamine (Invitrogen, 11668-019) and cells were plated in 24 well plates containing DMEM 18 hours prior to transformation. Then, 0.5 μg of total DNA was injected into each well and transformed. Renilla luciferase gene was used as a reporter gene. Phocolin and TPA (Phorbol-12-Myristate-13-Acetate) were purchased from Sigma (P8139).

Experimental and control preparation

The control group Mock incubated on DMEM medium containing 10% FBS, the Starv group raised in EBSS (Earl's Balanced Salt Solution) for 2 hours, the Baf A1 group treated with Bafilomycin 100 nM for 2 hours, and the ITM2A gene The inhibitory effect of ITM2A on the offspring of the transformed TIM2A group was observed by reporter assay.

RFP-GFP-LC3 was used to observe the promotion and inhibition of childhood action in each of the reporter assays. LC3 is a marker of child action, RFP is a red fluorescent protein, and GFP is a green fluorescent protein. Since both RFP and GFP are expressed in child cells, they appear yellow when observed with a confocal microscope, but in autolysosomes, GFP is degraded and only RFP is expressed and appears red.

Immunization and western blot

To perform the immunoprecipitation method, cells were cultured and cultured in a dissolution buffer (150 mM NaCl, 50 mM HEPES ([pH 7.5], 1% NP40) containing a protease inhibition cocktail (Roche, 11 863 153 001) The protein immunoprecipitated from the cell lysate with safety was used for immunoblot analysis. For protein immunoblot analysis, the polypeptide of the whole cell lysate was separated by SDS-PAGE, and the protein was separated on a nitrocellulose membrane filter (UVP, Upland, Calif.) And LAS4000 system (GE Healthcare, Calif., USA) were used for detection of the protein as primary antibodies diluted 1: 2000 or 1: 5000 through an enhanced chemiluminescence (GST) -ITM2A amino-terminal fragment (amino acid 1-120) fusion protein was used as the antigen source for the anti-ITM2A antibody. GREB and phosphorylated-REB About Antibodies were purchased from Cell Signaling Technology (9197, 9198), LC-3 antibody from Novus biological (NB100-2220), v-ATPase antibody (ATP6V0A4) from Aviva Systems Biology (OAAB02785) A190-125A).

Immunofluorescence  And Confocal  microscope

GFP-LC3 cells were cultured in a sterile glass cover slip. After the drug treatment, the cells were fixed with 4% paraformaldehyde. For immunostaining, cells were blocked with PBS containing 10% goat serum, stained with primary antibody diluted 1: 500 and then stained with Alexa 488-conjugated secondary antibody (Invitrogen, A-11001 ). Finally, the slides were washed three times with PBS, stained with DAPI and placed in mounting medium (Vector, H-1000). Images were taken with a Carl Zeiss LSM710 confocal microscope (Oberkochem, Germany). EEA1 antibody was purchased from BD (610457) and LAMP1 antibody was purchased from Santa Cruz (SC-20011).

Confirm Cancer Cell Growth Suppression

HeLa cells were cultured on 6-well plates and transformed with pcDNA3.1 vector and hPIPa (pcDNA3.1 vector containing ITM2A gene), respectively. Two days later, the transformed cells were cultured for 20 days under 1 mg / ml G418 treatment. Thereafter, the cells were fixed with methanol, stained with crystal violet, and colonies were observed.

In cancer cells ITM2A  Measurement of expression level

A microarray was used to confirm the reduction of expression of ITM2A in cancer cells. CDNA synthesized from cancer tissue and mRNA extracted from normal tissue was mixed with the microarray. The microarray scanner was then measured and quantified for binding on the microarray. The amount of expression of ITM2A gene in normal tissues was 1, and the remaining cancer cell tissues were expressed as relative values (see FIG. 6).

<Experimental Results>

ITM2A Confirming the inhibitory action of

RFP-GFP-LC3 was used in the present invention in order to examine whether the child's action is inhibited, LC3 is a marker of child action, and RFP is a red fluorescent protein. In addition, GFP is a green fluorescent protein. Since both RFP and GFP are expressed in child cells, they are yellow in the confocal microscope, but in autolysosomes, GFP is completely degraded and only RFP is expressed and appears red.

As shown in FIG. 2, when ITM2A is cloned into a plasmid and overexpressed, it can be confirmed that a large amount of autophagosome is formed. When cells expressing GFP-LC3 were starved (starved, left panel of Mock panel 2 and left panel Baf A1) and treated with Bafilomycin A1 (Baf A1) (Fig. 2 Baf A1 panel), the boll was formed.

The formation of large amounts of autophagosomes may be induced by lack of starvation energy in the cells, but it can be over-formed when Baf A 1 inhibits the parental flow. 2, it can be seen that the boll of ITM2A is very similar to that formed when Baf A1 inhibits the child's action

ITM2A Confirmation of Child Action Flow Control by

When the cells were starved to induce the child's autolysosome formation, red bafilomycin A1 treatment inhibited the formation of yellowish bolls. As shown in Fig. 3, it can be seen that ITM2A inhibits the flow of the child's action like Bafilomycin A1 and interferes with the child's action. When the cells were starved, autolysosomes were formed (Fig. 3, starv. B), but ITM2A expression was observed to form yellowish bolls like Baf A1 (Fig. 3 Baf A1. C, ITM2A. D ).

ITM2A Inhibition of cancer cell growth by

Because ITM2A inhibits the action of the child, cell growth can be inhibited during overexpression. The present inventors confirmed that ITM2A inhibits colony formation by observing colony formation after overexpression of ITM2A in cervical cancer cell line HeLa. From the experimental results, it was confirmed that the cancer cells did not grow well by ITM2A. As shown in FIG. 4, it was confirmed that colony formation was inhibited when ITM2A was overexpressed in the cervical cancer cell line HeLa cells (lower panel in FIG. 4) as compared with the control group (FIG. 4 upper panel).

In cancer cells ITM2A Confirmation of decreased expression of

The expression of ITM2A in normal tissues and ovarian cancer tissues was confirmed using a microarray. Green is normal tissue (control) and red is ovarian cancer (tumor) tissue. As shown in FIG. 5, it was confirmed that the ITM2A gene significantly decreased the expression level in cells such as ovarian cancer.

The ITM2A polypeptide of the present invention can be used as an effective ingredient to inhibit the child's action and further effectively kill cancer cells and the like. The composition of the present invention has the effect of inhibiting the child action, making the cancer cells more responsive to the anticancer agent, and effectively killing the cancer cells, which is highly industrially applicable.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Anti-tumor composition comprising ITM2A polypeptide or DNA coding          ITM2A gene <130> NP14-0056 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 263 <212> PRT <213> Artificial Sequence <220> <223> ITM2A peptide sequence <400> 1 Met Val Lys Ile Ala Phe Asn Thr Pro Thr Ala Val Gln Lys Glu Glu   1 5 10 15 Ala Arg Gln Asp Val Glu Ala Leu Leu Ser Arg Thr Val Arg Thr Gln              20 25 30 Ile Leu Thr Gly Lys Glu Leu Arg Val Ala Thr Gln Glu Lys Glu Gly          35 40 45 Ser Ser Gly Arg Cys Met Leu Thr Leu Leu Gly Leu Ser Phe Ile Leu      50 55 60 Ala Gly Leu Ile Val Gly Gly Ala Cys Ile Tyr Lys Tyr Phe Met Pro  65 70 75 80 Lys Ser Thr Ile Tyr Arg Gly Glu Met Cys Phe Phe Asp Ser Glu Asp                  85 90 95 Pro Ala Asn Ser Leu Arg Gly Gly Glu Pro Asn Phe Leu Pro Val Thr             100 105 110 Glu Glu Ala Asp Ile Arg Glu Asp Asp Asn Ile Ala Ile Ile Asp Val         115 120 125 Pro Val Pro Ser Phe Ser Asp Ser Asp Pro Ala Ala Ile His Asp     130 135 140 Phe Glu Lys Gly Met Thr Ala Tyr Leu Asp Leu Leu Leu Gly Asn Cys 145 150 155 160 Tyr Leu Met Pro Leu Asn Thr Ser Ile Val Met Pro Lys Asn Leu                 165 170 175 Val Glu Leu Phe Gly Lys Leu Ala Ser Gly Arg Tyr Leu Pro Gln Thr             180 185 190 Tyr Val Val Arg Glu Asp Leu Val Ala Val Glu Glu Ile Arg Asp Val         195 200 205 Ser Asn Leu Gly Ile Phe Ile Tyr Gln Leu Cys Asn Asn Arg Lys Ser     210 215 220 Phe Arg Leu Arg Arg Arg Asp Leu Leu Leu Gly Phe Asn Lys Arg Ala 225 230 235 240 Ile Asp Lys Cys Trp Lys Ile Arg His Phe Pro Asn Glu Phe Ile Val                 245 250 255 Glu Thr Lys Ile Cys Gln Asp             260 <210> 2 <211> 792 <212> DNA <213> Artificial Sequence <220> <223> ITM2A sequence <400> 2 atggtgaaaa tcgccttcaa tacccctacc gccgtgcaaa aggaggaggc gcggcaagac 60 gtggaggccc tcctgagccg cacggtcaga actcagatac tgaccggcaa ggagctccga 120 gttgccaccc aggaaaaaga gggctcctct gggagatgta tgcttactct cttaggcctt 180 tcattcatct tggcaggact tattgttggt ggagcctgca tttacaagta cttcatgccc 240 aagagcacca tttaccgtgg agagatgtgc ttttttgatt ctgaggatcc tgcaaattcc 300 cttcgtggag gagagcctaa cttcctgcct gtgactgagg aggctgacat tcgtgaggat 360 gacaacattg caatcattga tgtgcctgtc cccagtttct ctgatagtga ccctgcagca 420 attattcatg actttgaaaa gggaatgact gcttacctgg acttgttgct ggggaactgc 480 tatctgatgc ccctcaatac ttctattgtt atgcctccaa aaaatctggt agagctcttt 540 ggcaaactgg cgagtggcag atatctgcct caaacttatg tggttcgaga agacctagtt 600 gctgtggagg aaattcgtga tgttagtaac cttggcatct ttatttacca actttgcaat 660 aacagaaagt ccttccgcct tcgtcgcaga gacctcttgc tgggtttcaa caaacgtgcc 720 attgataaat gctggaagat tagacacttc cccaacgaat ttattgttga gaccaagatc 780 tgtcaagatt aa 792

Claims (12)

The antitumour cancer composition for inhibiting the cell function of a cell comprising the ITM2A polypeptide as an active ingredient to promote cell death.
The antitumor cancer composition according to claim 1, wherein the ITM2A polypeptide comprises the amino acid sequence of SEQ ID NO: 1.
delete delete The composition of claim 1, wherein the anti-cervical cancer composition is administered in combination with an anti-cancer agent.
6. The composition of claim 5, wherein the anti-cervical cancer composition is administered simultaneously, seperately or sequentially with an anti-cancer agent.
The pharmaceutical composition according to claim 5 or 6, wherein the anticancer agent is selected from the group consisting of fluorouracil, everolimus, metformin, doxorubicin, Epigallocatechin gallete (EGCG) Wherein the antitumor agent is at least one selected from the group consisting of trastuzumab, rapamycin, and cisplatin.
A composition for treating cervical cancer of the uterine cervix, comprising: a nucleic acid encoding the polypeptide of claim 1 as an active ingredient;
9. The composition of claim 8, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 2.
9. The composition of claim 8, wherein the nucleic acid is contained in an expression vector.
11. The composition of claim 10, wherein the expression vector is a plasmid or a viral vector.
12. The composition of claim 11, wherein the viral vector is selected from retroviral vectors, adenoviral vectors, and herpes virus vectors.

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