US20100272692A1 - Anticancer composition comprising plant stem cell line derived from taxus cambium or procambium - Google Patents

Anticancer composition comprising plant stem cell line derived from taxus cambium or procambium Download PDF

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US20100272692A1
US20100272692A1 US12/682,574 US68257408A US2010272692A1 US 20100272692 A1 US20100272692 A1 US 20100272692A1 US 68257408 A US68257408 A US 68257408A US 2010272692 A1 US2010272692 A1 US 2010272692A1
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cell line
procambium
cambium
cancer
extract
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Joong Hyun Park
Min Jung Lim
Il Seok Oh
Dae Hee Lee
Jung Chang Lee
Eun Kyong Lee
Young Woo Jin
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Unhwa Corp
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Unhwa Corp
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Publication of US20100272692A1 publication Critical patent/US20100272692A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/13—Coniferophyta (gymnosperms)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/04—Plant cells or tissues

Definitions

  • the present invention relates to a composition for preventing or treating cancer, which contains, as an active ingredient, a Taxus cambium- or procambium-derived cell line; a lysate thereof; an extract thereof; or a culture medium thereof.
  • Apoptosis is an active process of cell death that occurs through the regulation of various gene expressions and protein activities in cells according to programmed signals. Apoptosomes formed through this process are removed by the action of phagocytes such as the surrounding cells or as macrophages, such that they do not cause inflammation. Such apoptosis is frequently observed in various normal physiological processes of living organisms and known to be deeply involved in the process of development of various diseases. Namely, the development of abnormal apoptosis can lead to neurodegenerative disorders, immune disorders and cardiovascular diseases, and the abnormal inhibition of apoptosis can cause cancer.
  • apoptosis diseases which are caused by the abnormal development and inhibition of apoptosis include cancers induced by the abnormal expression of genes such as p53, p16 and Bc1-2, infections with HIV Herpes and influenza virus, and autoimmune diseases such as type 1 diabetes, rheumatoid arthritis, multiple sclerosis and myasthenia gravis.
  • the apoptosis of cells of an individual is a general mechanism for removing abnormal cells, that is, unrecoverable genetically damaged cells, from the individual in order to prevent the development of tumors caused by either the genetically damaged cells or the induction of inappropriate differentiation by differentiation stimuli.
  • the disturbance of apoptosis process induces the survival of damaged cells and cells that started to be damaged and the growth of these cells, and thus the inhibition of apoptosis plays an important role in a carcinogenic process.
  • substances having cancer preventive effects induce the apoptosis of such abnormal cells and that the induction of apoptosis by these substances is associated at least with the cancer preventive activity thereof (Fesus, L., J. Cell Biochem., 22:151, 1995; Reddy, B. S., Cancer Res., 57:420, 1997)
  • Some of the present inventors developed a method for providing cambium- or procambium-derived cell lines which overcome the problem of variation caused by dedifferentiation, can stably proliferate and has high genetic stability, and found that, when a Taxus -derived cell line among the cell lines is cultured, paclitaxel is obtained at a high yield (PCT/KR2006/001544).
  • paclitaxel is obtained at a high yield (PCT/KR2006/001544).
  • the anticancer effects of a Taxus cambium- or procambium-derived cell line itself have not yet been reported.
  • the present inventors have made extensive efforts to develop a natural material-derived anticancer composition, which has minimized side effects compared to the conventional anticancer agents and shows excellent anticancer activity, and, as a result, have found that a Taxus cambium- or procambium-derived homogeneous cell line; a lysate thereof, an, extract thereof; or culture medium thereof shows cancer cell-killing activity, thereby completing the present invention.
  • the present invention provides a pharmaceutical composition for preventing or treating cancer, which contains any one or more selected from the group consisting of a cell line, which is derived from the cambium or procambium of Taxus and has the following characteristics; a lysate thereof; an extract thereof; and a culture medium thereof:
  • the present invention provides a functional food for preventing or improving cancer, which contains any one or more selected from the group consisting of said cell line; said lysate; said extract; and said culture medium.
  • the present invention provides the use of any one among said cell line, said lysate; said extract; and said culture medium. to prevent or treat cancer.
  • the present invention provides a method for preventing or treating cancer, the method comprises using any one or more selected from the group consisting of said cell line; said lysate; said extract and said culture medium.
  • FIG. 1 depicts photographs showing a process of cell line induction according to the present invention, the image isolated procambium and cambium, and the comparison between dedifferentiated cells and Taxus procambium-derived cells.
  • FIG. 2 depicts comparative graphs of ROS amount and DCFH fluorescence photographs, which show the antioxidant activity of cell line extracts according to the present invention, when the aging of human skin diploid fibroblasts (HDFs) is induced by treatment with H 2 O 2 ,
  • p-WE procambium-distilled water extract
  • C-WE cambium-distilled water extract.
  • FIG. 3 depicts photographs showing the comparison of the size of cancer tissue between a control group and cell line-administered group at 2 weeks after administration of the cell line according to the present invention.
  • FIG. 4 is a graphic diagram showing the volumes of a control group and a cell line-administered group for 3 weeks after administration of a cell line according to the present invention.
  • FIG. 5 is a photograph of cancer tissues resected from mice of a control group and a cell-administered group at 3 weeks after administration of the cell line according to the present invention.
  • FIG. 6 is a photograph of the internal tissue of the resected cancer tissue at 3 weeks after administration of the cell line according to the present invention.
  • FIG. 7 depicts photographs showing the degree of cell death after the treatment of cancer cell lines with the cell line extract according to the present invention.
  • FIG. 8 is a photograph showing the degree of cell death after the treatment of a colon cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 9 is a photograph showing the degree of cell death after the treatment of a lung cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 10 is a photograph showing the degree of cell death after the treatment of a prostate cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 11 is a photograph showing the degree of cell death after the treatment of an osteosarcoma cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 12 is a photograph showing the degree of cell death after the treatment of an oral cavity carcinoma cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 13 is a photograph showing the degree of cell death after the treatment of a skin cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 14 is a photograph showing the degree of cell death after the treatment of a leukemia cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 15 is a photograph showing the degree of cell death after the treatment of a uterine cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 16 is a photograph showing the degree of cell death after the treatment of a skin cancer (human) cell line various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 17 is a photograph showing the degree of cell death after the treatment of a pancreas cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 18 is a photograph showing the degree of cell death after the treatment of a breast cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 19 is a photograph showing the degree of cell death after the treatment of a gastric cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 20 is a photograph showing the degree of cell death after the treatment of a renal cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 21 is a photograph showing the degree of cell death after the treatment of a liver cancer cell line with various concentrations of a methanol extract of the cell line according to the present invention.
  • FIG. 22 graphically shows the results of analysis of existence of paclitaxel in the cell line according to the present invention.
  • the term “cambium” refers to a tissue that thickens the stem and root to allow the plant grow volumetrically. It was reported that when the cambium, a meristem where the most active cell division occurs, is used as an explant for plant tissue culture, rapid and mass production of cells is possible (Korean Patent 10-0533120).
  • procambium refers to a primary meristem derived from an initial cell group, and the cambium that is a primary meristem is derived from the procambium without the intervene of permanent tissue.
  • lysate refers to a cell lysate obtained by disrupting cells through a chemical method with, for example, a detergent, or a physical method.
  • extract of a cell line refers to a substance obtained by dissolving cells in a solvent and isolating the cells, and the extract can be concentrated through distillation or evaporation.
  • the term “innately undifferentiated” means that cells are not present in an undifferentiated state through a dedifferentiation process, but are originally maintained in a pre-differentiated state.
  • the present invention relates to a pharmaceutical composition for preventing or treating cancer, which contains any one or more selected from the group consisting of a cell line which is derived from the cambium or procambium of Taxus ; a lysate thereof; an extract thereof; and a culture medium thereof:
  • the Taxus cambium- or procambium-derived cell line according to the present invention is additionally characterized in that: (a) it is present as single cells during suspension culture; (b) it has a higher growth rate than those of cell lines derived from tissues other than the cambium or procambium of Taxus and is cultured stably; and (c) it has low sensitivity to shear stress in a bioreactor compared to cell lines derived from tissues other than the cambium or procambium of Taxus.
  • the cell line according to the present invention is preferably obtained using an isolation method comprising the following steps of:
  • step (c) is preferably performed by proliferating the tissue in a medium, which contains 3-5 wt % of raw sugar or sugar and at least one substance selected from the group consisting of methyl jasmonate, fungal extract, bacterial extract, yeast extract, chitosan, glucomanan, glucan, phenylalanine, benzoic acid, salicylic acid, arachonic acid, STS, mevalonalonate N-benzolyglycine, ABA, SNP, IPP, BHT, CCC, ethephon, hippuic acid, amminoium ceric nitrate, AgNO 3 , vanadyl sulfate, p-aminobenzoic acid, brassinosteroids, sodium alginate, and sodium acetate, and then collecting the proliferated cell line.
  • methyl jasmonate is preferably contained in an amount of 10-100 ⁇ M.
  • the medium used in the present invention is a conventional medium for plant tissue culture, and examples thereof include, but are not limited to, N6 medium, SH medium, MS medium, AA medium, LS medium, B5 medium, WPM medium, LP medium, White medium, GD medium, DKW medium, DCR medium, etc.
  • the extract is preferably obtained using a solvent selected from the group consisting of distilled water, alcohol, acetone, DMSO (dimethyl sulfoxide), and mixed solvents thereof.
  • a solvent selected from the group consisting of distilled water, alcohol, acetone, DMSO (dimethyl sulfoxide), and mixed solvents thereof.
  • the Taxus cambium-derived cell line and procambium-derived cell line of the present invention had the effect of inhibiting reactive oxygen species induced by H 2 O 2 , suggesting that they had an antioxidant effect.
  • reactive oxygen species ROS
  • ROS reactive oxygen species
  • the Taxus cambium-derived cell line and procambium-derived cell line of the present invention were administered to mice, and then cancer tissues resected from the mice were observed.
  • the Taxus cambium-derived cell line and procambium-derived cell line of the present invention inhibited the growth of cancer tissues, inhibited angiogenesis as one of anticancer mechanisms, and enhanced the infiltration of immune cells into cancer tissue.
  • colon cancer, oral cavity carcinoma, lung cancer, prostate cancer, osteosarcoma, leukemia, uterine cancer, skin cancer, pancreas cancer, breast cancer, gastric cancer, renal cancer and liver cancer cell lines were treated with each of a distilled water extract, methanol extract and acetone extract of the cell lines in order to examine effects of the extracts on cancer cell death.
  • the cell line extracts according to the present invention were effective for the prevention and treatment of, but not limited to, colon cancer, oral cavity carcinoma, lung cancer, prostate cancer, osteosarcoma, leukemia, uterine cancer, skin cancer, pancreas cancer, breast cancer, gastric cancer, renal cancer and liver cancer.
  • the cell lines and extracts thereof. had activity for preventing and treating cancer.
  • a composition containing a lysate or culture medium of the cell line shows the effect of preventing and treating cancer
  • the composition containing the lysate or culture medium of the cell line according to the present invention can also show the effect of preventing and treating cancer.
  • the anticancer activity of the cell line of the present invention is not the action of paclitaxel known to be produced from Taxus , but is the anticancer activity of the cell line itself.
  • the cell line according to the present invention shows anticancer effects even when it is orally administered, the anticancer effect of the inventive cell line does not seem to be the action of paclitaxel which is used as an injection solution, because paclitaxel is ineffective when it is orally administered.
  • a composition for preventing or treating cancer containing any one or more of the cell line according to the present invention; a lysate thereof; an extract thereof; and culture medium thereof, may be provided as a pharmaceutical composition containing any one or more selected from the group consisting of the cell line; the lysate thereof, the extract thereof; and the culture medium thereof alone or in combination with at least one pharmaceutically acceptable carrier, excipient or diluent.
  • the cell line, the lysate, the extract, or the culture medium of the cell line may be contained as pharmaceutical composition in a pharmaceutically effective amount depending on disease and its severity, the patient's age, weight, health condition and sex, the route of administration and the period of treatment.
  • composition refers to a composition that is physiologically acceptable and does not cause gastric disorder, allergic reactions such as vertigo, or similar reactions, when administered to humans.
  • Examples of said carrier, excipient or diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oils.
  • the pharmaceutical composition may additionally contain fillers, anti-aggregating agents, lubricants, wetting agents, perfumes, emulsifiers and preservatives.
  • the pharmaceutical composition of the present invention may be formulated using a method well known in the art, such that it can provide the rapid, sustained or delayed release of the active ingredient after administration to mammals.
  • the formulation may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injection solutions, sterile powders, etc.
  • the present invention provides a functional food for preventing or improving cancer, which contains any one or more selected from the group consisting of said cell line; said lysate, said extract; and said culture medium derived from the cambium or procambium of Taxus.
  • the term “functional food” refers to a food, the functionality of which has been improved by adding the cell line according to the present invention, the lysate, the extract, or the culture medium of the cell line thereto.
  • the anticancer effect of the cell line of the present invention or the cell line extract may be used to prepare a functional food for preventing and alleviating cancer.
  • the functional food of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents, fillers, peptic acid and its salt, alginic acid and its salt, organic acids, protective colloid thickeners, pH regulating agents, stabilizers, preservatives, glycerin, alcohol, carbonizing agents that are used in carbonated beverages.
  • the plant was pretreated with a mixed solution of 1% benomyl (Dongbu Hannong Chemical, Korea), 1% daconil (Dongbu Hannong Chemical, Korea), 1% sterptomycin sulphate (DUCHEFA, The Netherlands) and 0.1% cefotaxime sodium (DUCHEFA, The Netherlands) for 24 hours, and then washed with tap water for 30 minutes to remove phenolic compounds and the remaining chemicals. Then, the plant was surface-sterilized in 70% ethanol (DC Chemical, Korea) for 1 min, 30% hydrogen peroxide (LG Chemical, Korea) for 15 min, 1% CLOROX solution for 15 min and 3% CLOROX solution for 5 min, and then washed 3-4 times with water.
  • the outer tissues of the twig and stem which have undergone the sterilization process, were readily peeled by pulling them in the lengthwise direction.
  • the peeled tissues were composed of xylem, procambium (twig) or cambium (stem), phloem, cortex and epidermis, and they were cultured in such a manner that the innermost tissue of the peeled tissues, that is, xylem, came into contact with a medium.
  • FIG. 1( b ) the top indicates a tissue containing phloem, cortex and epidermis, the middle indicates cambium, the bottom indicates xylem, and the arrow head indicates the separation between the cambium layer and the tissue containing phloem, cortex and epidermis.
  • “A” indicates a phloem/cortex/epidermis-containing tissue-derived cell line, which proliferated irregularly due to the difference in division between cells
  • “B” indicates a cambium-derived cell line, which proliferated to form a uniform cell layer through regular cell division
  • “C” indicates xylem in which cell division did not occur.
  • FIG. 1( c ) shows that a procambium-derived cell line is induced.
  • A shows the twig observed after 30 days of culture and is a photograph showing that the procambium (bottom) is separated from callus cells derived from the tissue composed of primary phloem, cortex and epidermis
  • B is a photograph after 35 days of culture and shows that the induced procambium layer was isolated and cultured
  • C is a photograph after 40 days of culture and shows that the callus of the tissue composed of primary phloem, cortex and epidermis proliferated after the isolation of the procambium layer.
  • the white and friable portion thereof having good growth rate was subcultured in the same fresh medium as induction medium at an interval of 21 days. Meanwhile, the medium used to induce only the procambium- and cambium-derived cell lines is shown in Table 1 below.
  • the growth regulator auxin was added to the medium at a concentration of 1-3 mg/L.
  • the culture was carried out in a dark room controlled at 25 ⁇ 1° C.
  • Taxus embryo and needle explants were sterilized, and then cultured in the medium of Table 1.
  • the callus induced from the embryo and needle explants had an irregular shape due to the difference in division rate between various cells like the case of the phloem-containing tissue, showed unstable growth rate and readily turned brown.
  • the brown and aggregated callus induced from the embryo and needle explants showed slow growth due to phenolic compounds secreted therefrom, and ultimately died. Namely, after 6 months of culture, the callus induced from the embryo and needle explants were difficult to maintain and culture.
  • the procambium- and cambium-derived cells were stably maintained without variations in their growth rate, growth pattern and aggregation, when they were cultured for a long period of more than 20 months, suggesting that the large scale cell culture was possible.
  • the procambium- and cambium-derived cell lines were placed in a flask containing a liquid medium shown in Table 2 below. Then, the cell lines in the flask were cultured in a rotating shaker at 100 rpm at 25 ⁇ 1° C. The subculture was set to 2 weeks, such that the cultured cells could always maintain high viability in the exponential growth phase.
  • embryo- and needle-derived callus were also cultured in medium 2 of Table 2 and compared with the procambium- and cambium-derived cell lines of the present invention.
  • the degree of aggregation of the cells was observed with biological microscope CX31 (Olympus, Japan). As a result, as shown in Table 3 below, it could be seen that more than 90% of cells of the cell lines according to the present invention were present as single cells upon suspension culture. As shown in FIG. 1( d ), it could be observed that the cell lines according to the present invention were morphologically characterized by a large number of vacuoles and were in an undifferentiated state. The arrow in FIG. 1( d ) indicates vacuoles in the Taxus procambium-derived cells.
  • the embryo/needle-derived callus and the procambium- and cambium-derived cells were cultured in an airlift bioreactor (Sung-Won Cytec, Korea) having an internal volume of 3 L.
  • the culture was carried out in the liquid medium of Table 2 under dark conditions at 25 ⁇ 1° C.
  • the cell lines according to the present invention showed about 2-3-folds higher growth rate compared to cell lines derived from tissues other than the procambium or cambium of Taxus , and when cultured in the bioreactor, the cell lines according to the present invention showed 5-6-folds higher growth rate compared to cell lines derived from tissues other than the procambium or the cambium. This is believed to be because cell viability rapidly decreased due to growth ring formation in the bioreactor, plant cell aggregation during culture, and the sensitivity of rigid cell walls to shear stress.
  • the procambium- and cambium-derived cell lines according to the present invention formed a very small growth ring area in the bioreactor, and the ring formed on the internal wall thereof was simply eliminated, when a simple stimulus was applied to the incubator to shake the medium. Also, it was shown that the inventive cell lines had low aggregation and contained a large number of vacuoles, and thus had low sensitivity to shear stress, such that cell viability did not decrease. In other words, it was seen that the cell lines according to the present invention had low sensitivity to shear stress resulting from shaking in the bioreactor for mass production, and thus could be produced rapidly in large amounts in the bioreactor.
  • Taxus procambium- or cambium-derived cell line had 2-3-folds lower sensitivity to shear stress compared to cell lines derived from tissues other than the procambium or cambium of Taxus .
  • the cell lines which have been suspension-cultured for 14 days as described in Example 1-4, were cultured in media (containing sterile water, 3-5 wt % (g/L) of raw sugar and 100 ⁇ M of methyl jasmonate) for 10 days in dark conditions, and then the cells were collected and used in the subsequent experiments.
  • media containing sterile water, 3-5 wt % (g/L) of raw sugar and 100 ⁇ M of methyl jasmonate
  • HDF Human Diploid Fibroblasts
  • HDF cells were isolated from the fetal penis prepuce and cultured.
  • the culture medium was prepared by adding 10% fetal bovine serum (FBS, Hyclone, Logan, Utah, USA) inactivated by heating at 56° C. for 30 minutes, 100 unit/ml of penicillin, 100 ⁇ g/ml of streptomycin and 300 ⁇ g/ml of glutamine to DMEM medium (Invitroge Gibco life tech. Vienna, Austria).
  • the cells were cultured in the medium, described above, in a 5% CO 2 incubator at a temperature of 37° C. and a humidity of 95% and subcultured at 3-4-day intervals, immediately before the cells were fused with each other.
  • the subcultured cells were divided, according to the number of subcultures (passages), into young cells cultured less than 20 passages, middle cells cultured for 21-49 passages, and aged cells cultured more than 50 passages.
  • 5 ⁇ 10 5 cells were dispensed into a 6-well plate, and then treated with H 2 O 2 alone or in combination with the extract obtained in Example 2.
  • the extract the distilled water extract among the extracts obtained in Example 2 was used at a concentration of 10-100 ⁇ g/ml, and preferably 50 ⁇ g/ml.
  • the cells were washed 2-3 times with HBSS (Hank's balanced salt solution) and stabilized in HBSS for about 30 minutes.
  • the cells were stained with 10 ⁇ M of DCFDA (Molecular Probes USA) in dark conditions at 37° C. for 1 hour, washed three times with HBSS, and then observed with a fluorescent microscope ( FIG. 2( c )).
  • the HDF cells were treated with 200 ⁇ M of H 2 O 2 and 10-100 ⁇ g/ml (preferably 50 ⁇ g/ml) of the distilled water extract obtained in Example 2, and the variation in the morphology of the cells was observed.
  • the HDF cells generate reactive oxygen species (ROS) by oxidative stress. Because non-fluorescent DCFDA is oxidized by reactive oxygen species to form DCF showing strong fluorescence, reactive oxygen species can be measured.
  • FACS Calibur Becton Dickinson Analytic Flow Cytometer, USA was used for measurement.
  • the Taxus cambium-derived cell line extract and the Taxus procambium-derived cell line extract all inhibited the production of reactive oxygen species (ROS).
  • ROS reactive oxygen species
  • P-WE procambium-distilled water extract
  • C-WE cambium-distilled water extract
  • Substances having excellent antioxidant activity are active substances that inhibit cellular damage caused by cellular oxidation, and reactive oxygen species (ROS) produced in vivo bind to cellular molecules, such as intracellular DNA, proteins and lipids, to cause cellular mutations, and are involved in the formation of cancer tissue by inhibiting the normal function of cells.
  • ROS reactive oxygen species
  • the Taxus cambium-derived cell line and procambium-derived cell line of the present invention had antioxidant effects
  • the Taxus cambium-derived cell line and procambium-derived cell line of the present invention have the effect of preventing cancer.
  • mice purchased from Damool Science (Daejeon, Korea) were bred according to general animal breeding regulations. Specifically, 6-week-old mice were purchased and adapted for about 5 days. Then, 1 ⁇ 10 6 CT-26 colon cancer cells (Korean Cell Line Bank KCLB80009) were injected subcutaneously into the right portion of the mouse back.
  • CT-26 colon cancer cells Korean Cell Line Bank KCLB80009
  • the growth of the cancer cells to a size approximately equal to the size of hulled millet was observed.
  • the each of the cambium-derived cell line and procambium-derived cell line obtained in Example 1 were freely fed to the mice.
  • a control group was fed with a conventional nutrient-containing mouse feed, and the administration of the cell lines was performed by finely crushing the same feed, adding the cell line thereto at a ratio of 1:1, preparing a feed having the same shape as that of the conventional feed, and allowing the mice free access to the prepared feed.
  • the average daily dosage of the cell line was 2-3 g of cell fresh weight/mouse.
  • each of the control group and the cell line-fed group consisted of 15 animals (7 animals for the cambium-derived cell line, and 8 animals for the procambium-derived cell line), and the experiment was repeated 3 times.
  • the size of cancer tissue in the model mice was observed. As a result, as shown in FIG. 3 , it could be observed that the growth of cancer tissue in the group administered with the cambium-derived cell line and the procambium-derived cell line was inhibited compared to the control group.
  • the volume of cancer tissue in the group administered with the cell lines was about 100 mm 3 , but the control group showed a cancer tissue volume which was more than 6-folds larger than that of the cell line-administered groups.
  • the cancer tissues were resected and photographed and, as a result, as shown in FIG. 5 , there was a significant difference in volume between the control group and the cell line-administered group.
  • the weight of the resected cancer tissues was measured and, as a result, it was observed that the average weight of the cancer tissues in the control group was 2.16 g, whereas the average weight of the cancer tissues in the cell line-administered group was 0.21 g which was about 1/10 of that of the control group. Accordingly, it could be found that the cell lines according to present invention inhibited the general growth of cancer cells, suggesting that these cell lines had the effect of preventing and treating cancer.
  • control group showed characteristics in that there was no region showing cell death called apoptosis, the tissue in the whole region was dense, and dividing cells (portions indicated by “b” in FIG. 6( a )) were large.
  • the fact that the dividing cells were large means that the cancer tissue in the control group continued to grow and expand.
  • tissue was not dense, and most cells showed the condensation of nuclei called apoptotic bodies (portions by indicated “c” in FIG. 6( b )) or division into various tissues. Such apoptotic bodies did not substantially appear in the control group.
  • erythrocyte portions indicated by “a” in FIG. 6( a )
  • the control group showed many blood vessels, but the cell line-administered group did not show blood vessels.
  • cells showing round nuclei predicted to be immune cells were significantly observed (portions indicated by “d” in FIG. 6( b )), but were not substantially observed in the control group.
  • Taxus cambium- or procambium-derived cell line of the present invention is involved directly in the growth of cancer to induce cell death, suggesting that it has the effect of preventing and treating cancer by inhibiting the general growth of cancer tissue.
  • it inhibits angiogenesis occurring in carcinogenesis and enhances the infiltration of immune cells into cancer tissue to induce strong immune activity, thus showing strong anticancer activity.
  • the degree of a decrease in cell growth in the group treated with the extract was compared with a control group treated with nothing, a vehicle group having methanol and acetone added thereto, and a positive control group treated with Taxus -derived paclitaxel (Sigma) as an anticancer agent.
  • the cells were washed twice with a cell culture medium warmed to 37° C. for a given time, and then 1 ml of MTT solution (5 mg/ml, without phenol red) was added thereto. Then, the cells were cultured again for 4 hours. The supernatant was removed, and the produced formazen precipitate was dissolved in 1 ml of DMSO and measured for absorbance at 570 nm. The number of survived cells was observed by fixing the cells with 50% methanol at 3 days after treatment with the extract and staining the adhered survived cell with crystal violet.
  • Taxus -derived cell lines from which the culture medium has been removed was dissolved in distilled water, methanol and acetone in sequence to obtain cell line extracts, and then cells were treated with each of the extracts.
  • the methanol extract showed the highest apoptotic activity in most of the tested cells, and this apoptotic activity was similar to the apoptotic effect in a positive control group (Taxol (paclitaxel) of FIG. 7 ) treated with about 10 ⁇ g/ml of paclitaxel.
  • the distilled water and acetone extracts also showed apoptotic effects, but showed apoptotic activity, which was generally lower than that of the methanol extract.
  • the vehicle control (vehicle FIG. 7 ) treated with only methanol and acetone had no effect on apoptosis. This suggests that the extracts of the cell lines contain substances showing cancer cell-killing activity.
  • the cells were treated with various concentrations of the methanol extract, and the degree of apoptosis of the cells was measured.
  • the methanol extract was dissolved in methanol at a concentration of 200 mg/ml and added to each of 1 ml of the test cell line culture medium at concentrations of 150, 400 and 800 ⁇ g/ml.
  • the growth of the cells was measured by the MTT method at a one-day interval from day 1 to day 3 after addition of the methanol extract, and at 3 days after addition of the methanol extract, the survived cells were fixed with methanol, and then stained with crystal violet.
  • the results of measurement for the 14 cancer cell lines are shown in FIG. 8 to FIG. 21 .
  • the methanol extract of the Taxus cambium- or procambium-derived cell line according to the present invention showed apoptotic activity in the 14 cancer cell lines in a concentration-dependent manner.
  • 800 ⁇ g/ml of the methanol extract showed strong apoptotic activity in all the cell lines, and this apoptotic activity was similar to the apoptotic activity of paclitaxel (10 ⁇ g/ml) used in the positive control group.
  • paclitaxel 10 ⁇ g/ml
  • the quantitative difference between paclitaxel used in the positive control group and the methanol extract of the cell line according to the present invention is attributable to the fact that paclitaxel is a highly purified substance, whereas the extract of the cell line is a mixture of a large number of compounds. Accordingly, it is considered that the apoptotic activity of the cell line according to the present invention will be further improved through a purification process.
  • a paclitaxel (Taxol) standard used in this Example was purchased from Sigma, and HPLC water (J. T. Baker), acetonitrile and methanol were filtered through a 0.2 ⁇ m filter and then used as a mobile phase.
  • Example 2 0.2 g of the cell line prepared in Example 1 was vortexed in 0.4 ml of MeOH for 5 minutes, and then extracted at room temperature for 1 hour. The extract was centrifuged at 13000 rpm for 5 minutes and filtered through a 0.2 ⁇ m filter, and the filtrate was quantitatively analyzed by UPLC.
  • the taxol standard was analyzed by UPLC at absorbances of 227 nm and 280 nm. As a result, as shown in FIG. 22( a ), the retention time was 4.67 minutes. In addition, it was reported that paclitaxel must exhibit a UV absorbance ratio (227 nm/280 nm) of about 0.05 (Castor & Tyler 1993), and the paclitaxel standard was measured in this Example and, as a result, the UV absorbance ratio, A (280 nm)/A (227 nm), was 0.048.
  • the cell line (sample) according to the present invention was analyzed by UPLC at absorbances of 227 nm and 280 nm. As a result, as shown in FIG. 22( b ), the retention time was 4.70 minutes.
  • the PDA spectrums of the paclitaxel standard and the cell line extract according to the present invention were analyzed. As shown in FIG. 22( c ) and FIG. 22( d ), the analysis results revealed that the cell line according to the present invention contained no paclitaxel.
  • 100 mg of the cell line extract prepared in Example 1 was mixed with 100 mg of maize starch, 100 mg of lactose and 2 mg of magnesium stearate, and the mixture was compressed into a tablet according to a conventional tableting method.
  • Example 1 1 g of the cell line prepared in Example 1 was mixed with 10 g of isomerized sugar, 5 g of mannitol and a suitable amount of purified water, and the mixture was prepared into 100 ml of a syrup formulation according to a conventional method.
  • Preparation 1 200 mg of the cell line prepared in Example 1 was dissolved in 96 ml of water, and then 500 mg of vitamin C as a supplement, 1 g of each of citric acid and oligosaccharide as flavor enhancers and 0.05 g of sodium benzoate as a preservative were added thereto. Then, purified water was added thereto, thus preparing 100 ml of a functional beverage.
  • Preparation 2 200 mg of the cell line extract prepared in Example 1 was dissolved in 96 ml of water, and then 500 mg of vitamin C as a supplement, 1 g of each of citric acid and oligosaccharide as flavor enhancers and 0.05 g of sodium benzoate as a preservative were added thereto. Then, purified water was added thereto, thus preparing 100 ml of a functional beverage.
  • a cell line according to the present invention and a lysate, extract and culture medium thereof are derived from a natural material, have minimized side effects compared to prior therapeutic drugs, are safe to the human body, are involved directly in the growth of cancer to induce cancer cell death, and show anticancer activity of inhibiting angiogenesis occurring in carcinogenesis. Accordingly, they are useful for the prevention, treatment and alleviation of cancer.

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US20080194025A1 (en) * 2005-10-31 2008-08-14 Unhwa Stability of Secondary Metabolite Mass Production Through Synchronized Plant Cell Cultures
US20090011477A1 (en) * 2005-10-31 2009-01-08 Unhwa Stability of Secondary Metabolite Mass Production Through Synchronized Plant Cell Cultures
US20100233813A1 (en) * 2007-09-21 2010-09-16 Unhwa Corporation Plant stem cell line derived from cambium of herbaceious plant with storage root and method for isolating the same
US20110117039A1 (en) * 2005-10-31 2011-05-19 Unhwa Corporation Antioxidant, anti-inflammatory or anti-aging composition containing taxus cambium- or procambium-derived cell line as active ingredient
US20110229443A1 (en) * 2008-11-06 2011-09-22 Unhwa Corporation Composition for Cancer Prevention or Treatment Containing as Active Ingredient Plant Stem Cell Line Dervied from Cambium of Panax Ginseng Including Wild Ginseng or Ginseng
US20140199341A1 (en) * 2010-01-19 2014-07-17 Chieh-Chou Yu Use of antrodia camphorata for treating diseases
US9289457B2 (en) 2008-08-14 2016-03-22 Unhwa Coproration Composition for preventing or treating liver diseases, containing plant stem cell lines derived from the cambium of Panax ginseng including mountain ginseng or ginseng as active ingredient
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US11872258B2 (en) * 2018-12-06 2024-01-16 Stemstix, Inc. Oral cavity treatments

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KR20100127727A (ko) * 2009-05-26 2010-12-06 주식회사 운화 버드나무과의 형성층 유래 식물줄기세포 및 이의 분리배양방법
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5310672A (en) * 1992-04-01 1994-05-10 Union Camp Corporation Induction of somatic embryogenisis in taxus, and the production of taxane-ring containing alkaloids therefrom
US5407816A (en) * 1992-02-20 1995-04-18 Phyton Catalytic, Inc. Enhanced production of taxol and taxanes by cell cultures of taxus species
US5850032A (en) * 1992-04-01 1998-12-15 Union Camp Corporation Method for production of plant biological products in precocious neomorphic embryoids
US7264951B1 (en) * 1992-02-20 2007-09-04 Phyton, Inc. Enhanced production of taxol and taxanes by cell cultures of Taxus species

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5019504A (en) * 1989-03-23 1991-05-28 The United States Of America As Represented By The Secretary Of Agriculture Production of taxol or taxol-like compounds in cell culture
KR950005081B1 (ko) * 1993-07-06 1995-05-18 산림청임목육종연구소 주목 종자의 씨눈 유래의 체세포배 및 체세포배성세포 혹은 그 배양배지로부터 택솔(Taxol) 및 그 유도체를 생산하는 방법
KR960008427B1 (en) 1993-07-19 1996-06-26 Lg Inf & Comm Ltd Multi-terminal connecting apparatus of pbx
KR970010602B1 (ko) * 1993-12-22 1997-06-28 주식회사 태평양 주목의 세포배양에 의한 택솔의 생산방법
KR0150347B1 (ko) * 1995-04-29 1998-10-15 박종헌 결정화에 의한 텍서스속 식물체로 부터 택솔의 정제방법
KR100533120B1 (ko) 2003-06-16 2005-12-14 주식회사 운화 바이오텍 형성층을 이용한 식물세포배양 방법
WO2007052876A1 (en) * 2005-10-31 2007-05-10 Unhwa Stability of secondary metabolite mass production through syncronized plant cell cultures

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5407816A (en) * 1992-02-20 1995-04-18 Phyton Catalytic, Inc. Enhanced production of taxol and taxanes by cell cultures of taxus species
US7264951B1 (en) * 1992-02-20 2007-09-04 Phyton, Inc. Enhanced production of taxol and taxanes by cell cultures of Taxus species
US5310672A (en) * 1992-04-01 1994-05-10 Union Camp Corporation Induction of somatic embryogenisis in taxus, and the production of taxane-ring containing alkaloids therefrom
US5850032A (en) * 1992-04-01 1998-12-15 Union Camp Corporation Method for production of plant biological products in precocious neomorphic embryoids

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Khosroushahi et al. "Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata," Cell Biology International, 30; (2006), pgs. 262-269. *

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US9415081B2 (en) 2008-09-30 2016-08-16 Unhwa Corporation Composition for preventing or treating AIDS containing plant stem cell line derived from cambium of Panax ginseng including wild ginseng or ginseng as active ingredient
US20110229443A1 (en) * 2008-11-06 2011-09-22 Unhwa Corporation Composition for Cancer Prevention or Treatment Containing as Active Ingredient Plant Stem Cell Line Dervied from Cambium of Panax Ginseng Including Wild Ginseng or Ginseng
US9314492B2 (en) 2008-11-06 2016-04-19 Unhwa Corporation Composition for cancer prevention or treatment containing as active ingredient plant stem cell line dervied from cambium of Panax ginseng including wild ginseng or ginseng
US20140199341A1 (en) * 2010-01-19 2014-07-17 Chieh-Chou Yu Use of antrodia camphorata for treating diseases
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WO2023225268A1 (en) * 2022-05-19 2023-11-23 The University Of Chicago Vanadyl sulfate compositions and methods of use therefor for the treatment and prevention of aging-related diseases and cancer

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