KR101430310B1 - Pharmaceutical composition for prevention or treatment of colorectal cancer comprising Juncus effecus L extract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating colorectal cancer, which comprises an extract of mustard seed extract. The extract of R. chinensis according to the present invention has excellent cytotoxicity, inhibition of cancer cell growth and cell division inhibition activity by cell cycle G2 / M arrest, and thus can be effectively used as a preventive or therapeutic agent for cancer diseases, especially colon cancer. In addition, the sea mustard extract is a natural product and provides low toxicity, stability, and high therapeutic effect.

Description

[0001] The present invention relates to a pharmaceutical composition for prevention or treatment of colorectal cancer,

The present invention relates to a pharmaceutical composition for preventing or treating colorectal cancer, which comprises an extract of mustard seed extract.

Unlike normal cells, which can regulate cell proliferation, differentiation and survival, cancer can proliferate indefinitely as a basic disorder of cell regulatory mechanisms and transfer to the surrounding normal tissues or organs to take away the life of the individual.

Cancer is the leading cause of death not only in Korea but also in the world population, and the number of cancer cases is increasing every year. Among them, colorectal cancer is a malignant tumor of the colon and rectum. In general, the incidence of colorectal cancer in advanced countries is high. In Korea, the incidence of colorectal cancer is increasing with the westernization of diet.

Conventional cancer treatment methods include surgical surgery, radiation therapy, and chemotherapy. Recently, biotherapy such as immunotherapy to treat tumors by immunotherapy has been applied. However, despite the technological advances in these therapies, the 5-year survival rate for colorectal cancer patients is 70.1%, and deaths from colorectal cancer continue to increase every year. This is because existing treatments can temporarily inhibit cancer growth by surgical removal of cancer and radiation therapy and anticancer drug administration, but invasion and metastasis from residual cancer cells after treatment can not be predicted And this type of solid cancer is highly resistant to anticancer drugs. Therefore, it is impossible to save the life of the patient by previous treatment methods such as anticancer drug therapy. Therefore, recent studies have been actively conducted to effectively inhibit the growth of colon cancer.

Currently, colon cancer treatment methods used in Korea are surgery, radiation therapy, and chemotherapy. However, there are disadvantages such as tolerance, side effects and considerable economic burden. Therefore, a lot of research has been carried out to develop new medicines capable of reducing side effects of anticancer drugs and effectively treating cancer, and researches for finding anticancer drugs from natural plant sources have been actively conducted. The development of anticancer drugs using natural resources has the advantages of low toxicity, high stability, and high therapeutic efficacy because it is a herbal medicine or food that has been used for a long time.

On the other hand, ground beetroot is a perennial herbaceous plant belonging to the excess sirloin, and the center has been used as a herb since ancient times. It grows in water and wetlands all over the country. It is also called 'raspberry' because its stem is white like human bones. It is called 'raspberry' in that it made the core of raspberry in the core of lantern. It is distributed in Korea, Japan, China, North America, etc. It is 1.2 ~ 1.5m in height, 2.5㎜ in diameter, and has about 10 nodes, and each node has roots. The epidermis of the stem is an anabolic tissue containing chlorophyll, and there is a thick cell layer in it. At the end of the stem, about 20 greenish brown small flowers are opened as gunshot wounds. Although the pharmacological action of Ischia spp. Is known to be analgesic, diuretic, and hemostatic, there is no research on anticancer activity.

The present inventors have search for a substance that fewer side effects to obtain an effective anti-cancer components in natural products, intended for Chinese medicine inhibit the growth of human colon cancer cell lines, rush (Juncus effusus L. ) extract inhibits cell division by arresting the G2 / M cell cycle and has an excellent anticancer effect against human colon cancer cell lines, thus completing the present invention.

It is an object of the present invention to provide a pharmaceutical composition comprising a raspberry extract which can prevent or treat colon cancer.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating colorectal cancer comprising extract of Juncus Effesus L.

As used in the present invention, the term " roots " means a plant belonging to the genus Juncus effuses L. and belonging to the stem of a raspberry. Other names include sirloin, red bean, lamb fillet, jasper candle, lantern or lemon candle, noodle-shaped long cylinder, outer surface is milky white or light yellowish white, pressed flat, I have. It has been used in the past for urine problems, such as sick symptoms during urination, edema caused by pyelonephritis, symptoms of chest tightness and sleepiness, infant depression, pneumonia caused by pneumonia, and laryngitis.

For the purpose of the present invention, the filamentous fungus is used for the prevention or treatment of colon cancer.

As used herein, the term "shoulder seed extract" refers to extracts from common mustard seeds which are ordinary extraction solvents known in the art, for example, water, C _{1 -4} alcohols such as methanol, ethanol, butanol, And a mixed solvent of water and a solvent. Preferably, the sourcherry extract of the present invention is a C ₁ -C ₄ alcohol extract, more preferably a methanol extract.

The extract may contain any one of the extract obtained by the extraction treatment, the diluted or concentrated liquid of the extract, the dried product obtained by drying the extract, or the adjusted product or the purified product.

The extraction may be performed using any one selected from the group consisting of room temperature extraction, hot water extraction, cold extraction, reflux extraction, ultrasonic extraction, and steam extraction, but is not limited thereto.

In one specific example, the extract of Rifle root extract according to the present invention was extracted with 100% methanol, and the filtrate was concentrated under reduced pressure to prepare a methanol extract of Rifle root (Example 1).

The rumen extract of the present invention can be used to prevent or treat abnormal or undesirable cell numbers, benign or malignant diseases characterized by cell growth or cell survival, such as cancer. The cancer may be, but is not limited to, colorectal cancer, breast cancer, lung cancer, prostate cancer, myeloid leukemia, cervical cancer, and the like.

Preferably, the rumen extract of the present invention is used for the purpose of preventing or treating colon cancer.

As used herein, the term "cancer" refers to a disease associated with the control of death of a cell, which is caused by excessive proliferation of cells when the normal apoptosis balance is broken. As used herein, the term "colon cancer" means a malignant tumor consisting of cancerous cells in the large intestine.

In a specific embodiment, the methanol extract of mustard seeds was treated with HT29, a human colon cancer cell line. As a result, the methanol extract of mustard seed had excellent cytotoxic effect on HT29 cells and inhibited the growth of HT29 cells in a concentration- . As a result, it has been confirmed that extreme proliferation of abnormal cancer cells can be suppressed by using the extract of roe lotion, and ultimately, prevention or treatment effect of colon cancer can be achieved.

Preferably, the extract of R. cinerea of the present invention prevents or treat colon cancer by inhibiting the division of the colon cancer cells. In addition, the cockroach extract of the present invention preferably has an activity of stopping the G2 / M cell cycle.

Cell proliferation is caused by a continuous flow of complex and diverse biochemical reactions, the combination of which is called the cell cycle. The cell cycle consists of G1, S, M, and G2 sequences, which consist of the S phase that synthesizes DNA, the M phase that divides the cells, the preparatory G1 phase and the G2 phase, (sub G1) group. There are three checkpoints to check the overall condition of the cell to see if it is ready for the next step in the cell cycle to proceed to the next step. The first step is to determine the size of the cells with the restriction point (initiation) that occurs in the G1 phase to determine whether the DNA is cloned. The second is the G2 / M metastasis, which confirms that DNA replication is complete before cell division. The third is the metaphase-anaphase transition in the M phase, which initiates cytoplasmic breakdown. The extract of R. rumen according to the present invention has an activity of arresting the G2 / M cell cycle among the checkpoints.

In one specific example, Flow cytometry analysis was performed to analyze the effect of the sour-rumen extract of the present invention on the cell cycle of the human colon cancer cell HT29 cells. As a result, it was confirmed that the frequency of cells corresponding to G2 / M period was increased from 16.58% to 63.24% at 80 μg / ㎖ when the extract was not treated. In addition, it was confirmed that the frequency of cells corresponding to G1 and S period was decreased depending on the treatment of Seaweed extract.

Furthermore, in another specific example, western blotting test and immunofluorescence staining were performed in order to determine at which stage of the G2 / M cell cycle the filamentous root extract according to the present invention was involved. As a result, it was possible to confirm that the extract of R. mori extract inhibited cell proliferation by inhibiting cytoplasmic division in the second half of mitosis through HT2 cell cycle, G2 phase.

As used herein, the term "prophylactic " means any act that inhibits or delays colon cancer by the administration of a composition comprising the rumen extract. The term "treatment" as used in the present invention means all the actions of improving or improving the symptoms of the disease by administration of the composition containing the water lily extract.

As a specific example, the pharmaceutical composition for preventing or treating colorectal cancer according to the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent.

Examples of pharmaceutically acceptable carriers include, but are not limited to, saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, And if necessary, may further contain other commonly used additives such as antioxidants and buffers. In addition, a target organ or tissue-specific antibody or other ligand can be used in combination with the carrier so as to specifically act on the target organ. The carrier, excipient, or additive may be any conventional formulation, and the carrier, excipient, or additive is not limited by the above examples.

The composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories or sterilized injection solutions according to a conventional method. In detail, when formulating, it can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like. Such solid preparations may be prepared by mixing the above composition with at least one excipient such as starch, calcium carbonate, sucrose, lactose, Can be prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the non-aqueous solution and suspension include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

In another one aspect, the invention rush (Juncus effecus L. ) extract of the present invention is administered to a mammal including a human in need thereof for the prophylaxis or treatment of colorectal cancer.

The preferred dose of the RBIC extract according to the present invention may be appropriately selected by those skilled in the art depending on the mammal's condition and body weight, the degree of disease, the drug form, the route of administration and the period of time. In addition, all modes of administration may be expected, including, but not limited to, oral, rectal, or intravenous, intramuscular, subcutaneous, intrauterine, or intracerebral injection.

In another one aspect, the invention rush (Juncus effecus L. ) extract of the present invention for preventing or ameliorating colorectal cancer. Preferably, the preventive or remedial action of the colorectal cancer may be due to the action of inhibiting the division of colorectal cancer cells, that is, stopping the G2 / M cell cycle.

The health functional food may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, alginic acid and its salts, Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, it may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. In addition, the health functional food may be in the form of any one of meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, Lt; / RTI >

In addition, the health functional food may further include food additives, and the suitability of the food functional food as a "food additive" Standards and standards.

Examples of the products listed in the above-mentioned "food additives" include natural products such as ketones, chemical products such as glycine, potassium citrate, nicotinic acid and cinnamic acid, coloring matter, licorice extract, crystalline cellulose, guar gum, Sodium laurate, sodium glutamate preparation, noodles-added alkaline agent, preservative agent, tar pigment preparation and the like.

The extract according to the present invention, which is added to foods containing beverages in the course of manufacturing the health functional food, can be appropriately added or decreased as needed.

The extract of R. chinensis according to the present invention has excellent cytotoxicity, inhibition of cancer cell growth and cell division inhibition activity by cell cycle G2 / M arrest, and thus can be effectively used as a preventive or therapeutic agent for cancer diseases, especially colon cancer. In addition, the rumen extract is a natural product and has an advantage of providing low toxicity, high stability, and high therapeutic effect.

1 is a graph showing a result of a WST test according to an embodiment of the present invention.
2 is a graph showing the results of cell survival rate test results according to an embodiment of the present invention.
FIG. 3 is a graph showing the results of quantitative analysis of cell cycle analysis results according to one embodiment of the present invention.
FIG. 4 shows the results of the cell death induction test by DAPI staining according to one embodiment of the present invention.
5 is a diagram showing a result of Western blotting according to an embodiment of the present invention.
6 is a graph showing the results of an immunofluorescence staining experiment according to an embodiment of the present invention.
FIG. 7 is a graph quantifying the results of an immunofluorescence staining experiment according to an embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided to further understand the present invention, and the present invention is not limited by the examples.

Example  One: rush  Preparation of extract

Rush used in the experiment (Juncus effusus L.) was purchased from about herbal, samples for anti-cancer mechanism study on HT29 cells in the rush is extracted three times while put in 1L 100% methanol ground rush 100g at 75 ℃ 3 sigan The resulting extract was filtered through Whatman No.2 paper (Whatman International LTD., England), and the filtrate was concentrated under reduced pressure on a rotary evaporator (45 ° C) to obtain 2.7 g of a solid. Hereinafter, the methanol extract (Juncus effusus L. MeOH extracts) was referred to as "JEME ".

Example  2: Cancer cell culture

HT29 human colon cancer cells were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). Cell culture was performed with 10% (v / v) fetal bovine serum (Omega Scientific Inc., Tarzana, CA, USA) and 10 U / ml penicillin in RPMI 1640 medium (RPMI 1640 L-glutamine, Mediateck Inc.) Was added, and the cells were cultured at 37 ° C in 5% CO ₂ Lt; / RTI >

Example  3: JEME Cytotoxicity measurement

WST assay was performed to confirm cytotoxic effect on HT29 cells treated with JEME. HT29 cells were seeded at a concentration of 1 × 10 ⁴ cells / well in a 96-well plate, cultured for 24 hours, treated with JEME at an initial concentration of 500 μg / well, Lt; / RTI > After 24 hours of reaction, 20 μl of Premix WST-1 solution (Cell Proliferation Assay System, Takara, Otsu, Shiga, Japan) was added and incubated for 30 minutes. The optical density was measured by an enzyme-linked immunosolvent (ELISA) Lt; / RTI > at 540 nm.

As a result, as shown in FIG. 1, as the concentration of JEME increased to 20, 40, 80 and 160 / / ml, the number of viable cells of HT29 cells decreased to 67.39, 50.88, 33.19 and 5.01% The effect was confirmed. In addition, the IC _{50 value} (50% inhibitory concentration for HT29) of JEME was as low as 41 μg / ml, confirming that JEME had a superior cytotoxic effect.

Example  4: Measurement of proliferation inhibitory effect of cancer cells

HT29 cells were cultured in a 60 mm cell culture dish at a concentration of 0.75 × 10 ⁶ cells / ml and cultured for 24 hours. The cells were treated with JEME (20, 40, 60, 80 μg / Respectively. Cells were collected and stained with trypan blue solution (0.4%, Sigma-Aldrich Co., St. Louis, Mo., USA) at day 0, 1, 2, 3, And the cell proliferation inhibitory effect was measured by comparing the mean number of viable cells in the control group and the mean number of viable cells in the sample treated group.

As a result, as shown in FIG. 2, the survival rate of the cell line treated with JEME was 50.23% lower than that of the control group from day 1, and the survival rate was lower as the concentration of JEME increased.

Also, as the treatment time of JEME increased, the survival rate of HT29 cells decreased and the cell viability was about 3.43% when treated with 40 ㎍ / ㎖ of JEME for 5 days. This confirms that JEME inhibits the growth of HT29 cells in a concentration- and time-dependent manner.

Example  5: Flow cytometry Cell cycle analysis using

In order to investigate whether the proliferation inhibition of HT29 cells by JEME is related to the disturbance of specific cell cycle, JEME was treated with concentration (0, 20, 40, 60, 80 ㎍ / ㎖) for 24 hours and then flow cytometry The frequency of each cell was quantitatively analyzed.

Specifically, to determine the effect of JEME on the cell cycle of HT29 cells, HT29 cells were cultured in a 60 mm cell culture dish at a concentration of 0.75 × 10 ⁶ μg / ml and cultured for 24 hours. , 80 / / ml), and the control group was treated with 0.1% DMSO. After 24 hours, the cells were collected, washed with PBS, and stained with Cycle TEST PLUS solution. Cell cycle analysis was performed by flow cytometry (FACS Caliber, Becton Dickinson, San Jose, CA) after staining DNA using CycleTEST ™ PLUS KIT (Becton Dickinson, Heidelberg). CellQuest software; Becton Dickinson, and the results are shown in Table 1 and FIG.

Concentration of roe extract (㎍ / ㎖) Apoptotic cells
(Sub G1) Non-apoptotic cells G1 S G2 / M 0 1.48 66.78 17.06 16.58 20 1.99 62.33 19.32 18.71 40 2.86 62.85 14.53 23.11 60 2.82 36.04 10.57 53.97 80 2.99 32.37 3.92 63.24

As a result, as shown in Table 1 and FIG. 3, when the JEME treatment was not performed, the frequency of cells corresponding to the G2 / M period was increased in a dose dependent manner with the treatment of JEME at about 16.58% %, Respectively. In addition, it was confirmed that the frequency of cells corresponding to G1 and S period was decreased in a concentration dependent manner by treatment with JEME.

On the other hand, sub-G1 cells, which can be assumed to be apoptosis-inducing, did not change even when JEME treatment concentration was increased.

Example 6: Confirmation of cell death induction of HT29 cells

In order to confirm whether JEME induces apoptosis induction in HT29 cells, nuclear segregation specifically observed in apoptotic cells was confirmed by DAPI staining. Specifically, HT29 cells were treated with JEME at different concentrations (0, 20, 40, 60 ug / ml) and then stained with DAPI, a fluorescent material that specifically binds to nucleic acids, and observed using a fluorescence microscope.

As a result, as shown in FIG. 4, apoptotic body formation was not observed in HT29 cells treated with JEME, and thus JEME did not induce apoptosis in HT29 cells. As a result, it was confirmed that JEME inhibits cell proliferation by stopping the cell cycle in the G2 / M group in HT29 cells.

Example  7: Western Blot  analysis

The cell cycle is regulated by the activation of various Cdk and cyclin. Activation or inactivation of the cyclin / cyclin-dependent kinase (Cdk) complex depends on the cell cycle and the intracellular and extracellular signals, Or induction of differentiation and induction of cell death by apoptosis. Cdc2, a Cdk protein that functions in the G2 / M phase, is activated by binding to CyclinB or by dephosphorylation of the Tyr15 residue by Cdc25c. Activated Cdc2 regulates the progression of the M group by inducing phosphorylation of histone and lamin proteins and promoting nuclear decay and chromosome formation.

Through the above-described flow cytometry cell cycle analysis, HT29 cell proliferation inhibition was directly related to G2 / M arrest. Specifically, in order to confirm which protein acts to stop the cell cycle in the G2 / M phase, the expression change of the protein acting in the G2 / M phase was confirmed by Western blot analysis.

Specifically, HT29 cells were cultured in a 100 mm cell culture dish at a concentration of 2 × 10 ⁶ cells / ml, cultured for 24 hours, treated with JEME (20, 40, 60, 80 μg / DMSO. After incubation for 24 hours, cells were collected, washed once with PBS, and then treated with 0.1% Triton (Sigma) in CSK buffer (10 mM PIPES (pH 6.8), 100 mM NaCl, 1 mM MgCl ₂ , 1 mM EGTA, 1 mM dithiothreitol (DTT), 1 mM phenyl methane sulfonyl fluoride) The cells were suspended in a solution containing X-100, 1 mM ATP and 1% Protease inhibitor cocktail (BD Pharmingen, San Diego, CA), reacted for 15 minutes, and disrupted with an ultrasonic disintegrator. The disrupted cells were centrifuged at 14,000 rpm for 20 minutes. Then, proteins in the supernatant were quantified with a BCA protein assay kit (Bio-Rad, Hercules, Calif.) And 30 μg of the protein was dissolved in sodium dodecyl sulfate (SDS) And electrophoresed on amide gel. After electrophoresis, the protein in the gel was transferred to PVDF (Pall Corporation, Ann Arbor) membrane and blocked with 0.15 M NaCl, 1 M Tris-HCl (pH 7.5), 0.1% Triton X-100, 0.0 > 4 C < / RTI > for 16 hours. The primary antibody was reacted at 4 ° C for 20 hours. The membrane was washed with TBS buffer (50 mM Tris-HCl (pH 7.5), 0.15 M NaCl, 0.1% Triton X-100) Lt; 0 > C for 16 hours. After washing the membrane, the immunoreactive protein was detected by a chemiluminescence system (Super Signal West Femto Maximum Sensitivity Substrate, Pierce). The degree of the reaction was quantitated using FluorChem 占 5500 (Alpha Innothch, San Leandro, Calif.). The primary antibody used in this experiment was purchased from Santa Cruz Biothechnology Inc.

As a result, as shown in FIG. 5, after JEME treatment, Cdc25C tended to decrease, Cdc2 tended to decrease, but it was almost constant and Cyclin B1 tended to increase.

The activity of Cdc2 is affected not only by its binding to Cyclin B1 but also by the degree of phosphorylation of Cdc2 itself, which is involved in the degree of phosphorylation of Cdc2. Cdc25c is a phosphatase that dephosphorylates Cdc2 and promotes cell cycle from G2 to M group. Thus, from the results of FIG. 5, it can be seen that JEME induces inactivation of Cdc2 with the decrease of Cdc25c, which is similar to the results of previous studies related to cell cycle arrest in most G2 / M phases there was.

However, in the case of Cyclin B1, the expression level of Cyclin B1 is mostly decreased in the previous studies related to the cell cycle arrest in the G2 / M period, but the expression level is increased in HT29 cells treated with JEME. In the case of JEME treatment, the HT29 cells pass through the G2 phase and become mitosis, which is the result of the fact that Cyclin B1 is synthesized in S phase and activated in M phase, and is degraded by ubiquitination by APC (anaphase promoting omplex) In the second half of the study.

Example  8: Immunofluorescence  process of dyeing( Immunofluorescence staining )

As a result of JEME treatment, the expression of proteins involved in G2 / M regulation was observed to be somewhat decreased, but the expression of Cdc2 was increased in Cyclin B1. These results suggest that Cdc2 is activated by the increase of Cyclin B and that it passes through M group. Therefore, JEME is treated by concentration, and immunofluorescence staining is used to stain cytoplasm and nucleus, And to observe at which stage of M stage HT29 cells treated with JEME were located.

HT29 cells were cultured in a 35 mm culture dish with cover glasses, treated with JEME (20, 40, 60, 80 μg / ml) and treated with 0.1% DMSO. After incubation for 24 hours, HT29 cells were washed once with PBS, 1 ml of 4% formaldehyde was added, and the cells were reacted for 10 minutes. Then, the cells were washed three times with PBS, treated with PBS containing 0.2% triton X-100, It was permeated for 10 minutes. Washed three times with PBS and blocked with PBS containing 3% BSA for 1 hour at room temperature. The cells were then incubated at 4 ° C for 24 h, washed three times with PBS, and then incubated with donkey anti-free (Sigma Chemical Co., USA) Secondary antibody was diluted 1: 100 with IgG-FITC (Santa Cruz Biotechnology, Inc., San Diego, Calif., USA), shaded at room temperature and reacted for 2 hours. After washing with PBS 3 times, DAPI (Santa Cruz Biotechnology, Inc., San Diego, Calif., USA) reagent was treated with 4 μg / ml for 20 minutes and washed 3 times with PBS. Reactive HT29 cells were obtained from Axio Scope. Were taken using an A1 microscope (Carl Zeiss, Inc., Oberkochen, Baden-W, Germany).

As a result, as shown in FIG. 6 and FIG. 7, it was observed that as the treatment concentration of JEME increased, the rate of the nucleated cells increased as the cytosol separation step was stopped. From the above results, the number of cells that were not observed in the cytoplasmic cleavage step with respect to the total number of cells in each concentration group was measured and expressed as a percentage. As a result, the cells treated with 20 ㎍ / ㎖ of JEME showed similar morphology to those of the cells treated with 40 ㎍ / ㎖ of JEME. However, as the treatment concentration of JEME increased, Cells were confirmed to have increased. Therefore, it was confirmed that JEME inhibits cell proliferation by inhibiting cytoskeletal division in HT29 cells in the latter half of mitosis.

Claims (5)

A pharmaceutical composition for preventing or treating colorectal cancer, comprising a water extract of Cichoraceae.
The method of claim 1, wherein the rush extract is C ₁ -C ₄ alcohol to extract the composition. 3. The composition of claim 2, wherein the sea mustard extract is a methanol extract.
2. The composition according to claim 1, wherein the extract of Cryptomeria japonica protects or treats colon cancer by inhibiting colon cancer cell division.
The composition according to claim 1, wherein the sirloin extract has an activity of stopping the G2 / M cell cycle.

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