KR20220122914A - Composition for treatment of lung cancer including extract of beet as an effective ingredient - Google Patents

Abstract

The present invention relates to a composition for treating lung cancer containing a beet extract as an active ingredient. In addition, the beet extract is the composition for treating lung cancer, characterized in that there is a riboflavin-derived active oxygen inhibitory activity. Through the present invention, it is possible to provide the beet extract having antioxidant or anti-inflammatory effects.

Description

Composition for treatment of lung cancer including extract of beet as an effective ingredient

The present invention relates to a composition for treating lung cancer comprising beetroot extract as an active ingredient.

The most serious difficulties in patients with advanced cancer are symptoms due to cachexia, which is a major cause of death in cancer patients. In the case of patients receiving chemotherapy, the side effect of treatment is an excess of inflammatory substances (cachexia) due to damage to normal cells and destruction of cancer cells, and about 80% of cancer patients are experiencing it. Excessive cachexia is the biggest cause of suffering and shortening the lifespan of patients. The cause of cachexia is mainly an excess of cytokines, which are inflammatory substances, and the continuous expression of inflammation is pointed out as the biggest problem. They account for about 80% of the causes of death in cancer patients, and the persistent inflammation and pain are resolved. It is a matter to be dealt with.

On the other hand, oxidative stress (Oxidative stress) plays an important role in numerous physiological and pathological phenomena, as reactive oxygen species (ROS) accompanying oxidative stress include oxygen (O ₂ ), peroxidation ( superoxide, O ^- ), hydroxyl radical (HO ^- ), hydrogen peroxide (H ₂ O ₂ ), and the like. These reactive oxygen species are very reactive and cause DNA denaturation, excessive signal transduction and protein denaturation in the body. These harmful reactions are inhibited by antioxidants or antioxidant enzymes present in the living body, and are designed to maintain homeostasis. However, senescence of the antioxidant system according to aging and accumulation of free radicals due to continuous harmful stimuli break homeostasis and promote aging, and cause various diseases such as skin diseases, skin cancer, arteriosclerosis and blood clots (Laure Rittie et al. , Aging Research Reviews, 1, 705-720, 2002; Cutler RG, Annals of the New York Academy of Sciences, 1055, 93-135, 2005).

Inflammatory response is one of the body's defense responses to certain stimuli, and refers to a complex lesion that combines three types: tissue deterioration, circulatory disorder and exudation, and tissue proliferation. In addition, it can be said that it is the expression of a defense mechanism in vivo against various types of infection or irritants in in vivo metabolites, and various chemical mediators are involved in the expression mechanism of inflammation, and the pathogenesis is very complicated. It is a local protective reaction induced by tissue injury or destruction, and acts to destroy, weaken, or mask both the injury-causing agent and the injured tissue. The hallmark of such inflammation is that microvessels are perforated, blood components leak into the interstitial space, and leukocytes migrate to the inflamed tissue, which is usually accompanied by clinical symptoms such as erythema, edema, hyperalgesia and pain.

Various biochemical phenomena are involved as the cause of inflammation in the living body. It is known to play an important role in mediating. Accordingly, NOS, an enzyme that produces NO from L-arginine, or COX, an enzyme involved in synthesizing prostaglandins from arachidonic acid, plays a major role in blocking inflammation. According to recent research results, NOS exists in the brain bNOS (brain NOS), nNOS (neuronal NOS) present in the nervous system, eNOS (endothelial NOS), etc. present in the vascular system, and is always expressed at a certain level in the body. and NO produced in small amounts by them plays an important role in maintaining normal body improvement, such as inducing neurotransmission or vasodilation. On the other hand, NO, which is rapidly excessively generated by iNOS induced by various cytokines or external stimulants, is known to cause cytotoxicity or various inflammatory reactions, and there are studies that chronic inflammation is related to an increase in iNOS activity. Multifunctional cytokines such as TNF- are not only expressed in normal tissues, but their expression level is increased in the course of lesions, and in particular, plays an important role in skin inflammation that occurs in the process of promoting cancer. It has already been reported that TNF-rk is associated with human inflammatory skin diseases.

In addition, symptoms were alleviated when an antibody against TNF- was treated for various inflammatory diseases and allergic phenomena, and TNF- activates neutrophils and increases the production of hydrogen peroxide, thereby functioning as an endogenous cancer promoter. Therefore, by inhibiting the expression of TNF-, a cytokine that plays a pivotal role in the inflammatory stage closely related to the stage of promoting carcinogenesis, or by inhibiting the production of prostaglandin PGE 2 due to inhibition of COX-2 activity, the increase of the initial inflammatory molecules is prevented. It is known that there is a high possibility of controlling the concomitant lesion process. That is, when treated with antibiotics under bacterial infection, the LPS released from the outer membrane while killing the bacteria causes an endotoxin shock, and when it is severely destroyed by this process, it is not possible to neutralize the continuously generated LPS.

On the other hand, beets are known to have anticancer effects. It has been reported that beetroot extract can prevent breast and prostate cancer. It is known that a component called betalain contained in beetroot exhibits anticancer activity. Numerous studies support the anticancer and anticancer properties of beetroot. In particular, lettuce is a good source of functional substances with antioxidant action, such as betalains, flavonoids, polyphenols, vitamins, and folic acid. In particular, betalain, a pigment material in the root of red beet, is composed of betacyanin, a red violet pigment, and betaxanthin, a yellow pigment. Betanine, the main substance of betalain, is composed of a phenolic group and a cyclic amine group with excellent electron donating ability.

1. Republic of Korea Patent Publication No. 10-2016-0046782 2. Republic of Korea Patent Publication No. 10-2017-0080336

1. Jun Ho Lee and Koo Bok Chin. Evaluation of Antioxidant Activities of Red Beet Extracts, and Physicochemical and Microbial Changes of Ground Pork Patties Containing Red Beet Extracts during Refrigerated Storage. Korean J. Food Sci. An. 32(4), 497-503 (2012) 2. Mi-Ran Yi1, Kang Chang-Hee1,2, Hee-Jung Bu. Antioxidant and anti-inflammatory activity of extracts from red beet(Beta vulagaris) root. Korean J. Food Preserv. 24(3), 413-420 (2017) 3. Jung-Yun Kim, Hyun-Ku Kim. Physiological Activity of Redbeet. Bulletin of Food Technology. 22(3), 537-543 (2009)

An object of the present invention is to provide beetroot extract as an active ingredient having a therapeutic effect on lung cancer.

The present invention is an anti-inflammatory composition comprising beetroot extract as an active ingredient.

In addition, the beet extract is a composition for the treatment of lung cancer, characterized in that it has the ability to inhibit riboflavin-derived free radicals.

In addition, the beet extract is a composition for treating lung cancer, characterized in that it has NO production inhibitory ability.

On the other hand, in the present invention, the beet extract refers to the beetroot extract as water, methanol, ethanol, lower alcohol having 1 to 4 carbon atoms such as butanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N, Extracts obtained by leaching using N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof, carbon dioxide, pentane, etc. It refers to the obtained extract or a fraction obtained by fractionating the extract, and the extraction method may apply any method such as chilling, reflux, heating, ultrasonic radiation, supercritical extraction, etc. can In the case of the fractionated extract, it includes a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and sequentially fractionating the extract using the solvents in the order of increasing or decreasing polarity. It includes fractions, and further includes fractions obtained by chromatography using properties such as size, charge, hydrophobicity, and affinity. In addition, the meaning of the extract includes a concentrated liquid or solid extract from which the extraction solvent is removed by methods such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. When referring to the examples below, preferably an extract obtained by leaching using water, ethanol, or a mixed solvent thereof as an extraction solvent (especially a mixed solvent of water and ethanol), a high obtained by removing the extraction solvent from the extract It refers to the fraction of each layer obtained when the extract of the shape is suspended in water and sequentially fractionated with hexane, ethyl acetate and butanol. Here, 'sequential fractionation' means fractionation with the fractionation solvent in the order listed above by continuously using the residual water layer after fractionation.

The composition of the present invention may include the active ingredient in any amount (effective amount) as long as it can exhibit the ameliorating activity of the inflammatory disease intended for treatment depending on the use, formulation, compounding purpose, etc., but a typical effective amount is the total weight of the composition It will be determined within the range of 0.001 wt % to 15 wt % based on Herein, the term "effective amount" refers to an amount of an active ingredient capable of inducing improvement, treatment, or suppression/delay of the onset of such pathological symptoms in mammals, preferably humans, to which it is applied. Such effective amounts can be determined empirically within the ordinary ability of one of ordinary skill in the art.

Subjects to which the composition of the present invention can be applied (prescribed) are mammals and humans, particularly preferably humans. The composition of the present invention may be used as a pharmaceutical composition in a specific embodiment. The pharmaceutical composition of the present invention includes oral dosage forms (tablets, suspensions, granules, emulsions, capsules, syrups, etc.), parenteral dosage forms (sterile aqueous or oily suspensions for injection), topical formulations (solutions, creams, ointments, gels, lotions, patches), and the like. In the above, "pharmaceutically acceptable" means that it does not inhibit the activity of the active ingredient and does not have a toxicity (sufficiently low toxicity) beyond which the subject of application (prescription) can be adapted.

Through the present invention, it is possible to provide a beet extract having an antioxidant or anti-inflammatory effect.

1 is a graph showing the antioxidant activity of the beet extract fraction by the DPPH radical scavenging method.
2 is a graph showing the superoxide inhibitory activity of the beet extract fraction.
Figure 3 shows the activity of increasing the activity of the antioxidant enzyme of the beet extract fraction.
Figure 4 shows the active oxygen species reduction effect of the beet extract fraction.
5 shows the effect of increasing the expression of SOD-3 in transformed nematodes of the beet extract fraction.
6 shows the effect of increasing oxidative stress resistance of the beet extract fraction.
7 to 14 are graphs showing the antioxidant effect of beet extract.
15 to 16 are graphs showing the lung cancer treatment effect of beet extract.

Hereinafter, the present invention will be described with reference to specific examples.

The terms or words used in the present specification and claims are not to be construed as being limited in their ordinary or dictionary meanings, and the inventor may properly define the concepts of the terms to best describe his invention. Based on the principle, it should be interpreted as meaning and concept consistent with the technical idea of the present invention. Therefore, since the embodiments described in this specification are only the most preferred examples of the present invention and do not represent all the technical ideas of the present invention, there may be various equivalents and modifications that can be substituted for them at the time of the present application. It should be understood that there is

<Evaluation of antioxidant power of beets>

1. Materials and test methods

- Extraction method

After shredding the beets, 3.2 Kg of the sample obtained was extracted twice at 50°C for 5 hours while shaking with ethanol. The extract was concentrated under reduced pressure on a water bath to obtain about 164.93 g of ethanol extract, suspended in distilled water in this ethanol extract, and the same amount of n-hexane (7.71 g), methylene chloride (2.01 g), ethyl acetate (1.18 g) according to a conventional method. ) and n-butanol (2.68 g) were subjected to solvent fractionation in the order to obtain each fraction.

- Antioxidant activity by DPPH free radical scavenging method

In a 96-well plate, 0.2 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) (EtOH) was added in a certain amount to a solution prepared at each concentration with EtOH. After shaking for 10 seconds, it was left at 25oC for 30 minutes, and then absorbance was measured at 517 nm using a microplate reader. As a control drug, L-ascorbic acid was used. The antioxidant effect was graphically compared with the absorbance of the control group to which the sample was not added. DPPH radical scavenging activity for each sample was measured three times repeatedly.

- Riboflavin-derived superoxide inhibitory activity

The superoxide inhibitory ability of the sample is to measure the photochemical activity using an evaluation system composed of methionine, riboflavin, and NBT. The reaction mixture consisted of 2.6 μM riboflavin, 3 mM methionine, 75 μM NBT, 0.1 mM EDTA, PBS (pH 7.4), and samples of various concentrations. The mixture was placed in a light box and kept for 15 minutes, changing the position randomly every 5 minutes. The temperature inside the light box was maintained at 20±1oC, and the brightness of the light was maintained at 5,500 lux. NBT was reduced to blue formazane under light, and the product was measured at 560 nm. Inhibition of blue formazane formation becomes the ability to inhibit superoxide.

Cultivation of C. elegans - C. elegans was cultured on a Nematode Growth Medium (NGM) agar plate coated with E. coli OP50 at 20°C. Beet ethanol extract was added to sterilized NGM plates (at 50°C) as a stock solution using DMSO as a solvent. The final DMSO concentration was maintained at 0.1% (v/v) in all conditions.

- Measurement of antioxidant enzyme (SOD, catalase) activity in nematodes

N2 nematodes of the same growth stage were cultured on plates prepared by concentration of samples. On the 4th day after becoming adults, the nematodes were collected, washed three times with M9 buffer, and then pulverized and used for enzyme activity measurement (homogenization buffer: 10 mM Tris-HCl, 150 mM NaCl, 0.1 mM EDTA, pH7.5). SOD activity was measured by applying the method of Ibrahim et al. First, a reaction mixture (1.6 mM xanthine and 0.48 mM NBT 0.49 mL) was prepared with 10 mM phosphate buffer (pH 8.0) as a solvent, and then pre-incubated with 10 μL of each concentration sample at 37 oC for 5 minutes. After that, 100 μL (0.05 U/mL) of xanthine oxidase was added, incubated at 37 oC for 20 minutes, the reaction was stopped with 69 mM SDS, and absorbance was measured at 570 nm. Catalase activity was determined by applying Aebi's method to reacting 50 μL of each concentration sample in 25 mM H2O2 for 3 minutes and measuring the absorbance at 240 nm.

- Analysis of reactive oxygen species (ROS) in nematode cells

Reactive oxygen species in nematode cells were measured using 2',7'-dichlorodihydro fluorescein diacetate (H2DCF-DA). Nematodes having the same growth stage were cultured on plates prepared by sample concentrations. On the 4th day after they became adults, they were placed in M9 buffer containing 50 μM juglone, exposed for 2 hours, and then transferred 5 each to 50 μL M9 buffer contained in a 96 well plate. Finally, after adding 50 μL of 25 μM H2DCF-DA, absorbance was measured at excitation 485 nm and emission 535 nm, respectively.

Measurement of SOD-3::GFP fluorescence in transformed nematodes- As transformed nematodes, CF1553 containing SOD-3::GFP was cultured in a medium administered by concentration. It was used on the 4th day after becoming an adult, and the nematodes were anesthetized with sodium azide (4%), and the expression of GFP was observed with a fluorescence stereo microscope (Olympus, Japan). In order to quantify and analyze the expression intensity, photos were taken using a microscope and analyzed using ImageJ software. All experiments were repeated three times.

- Statistical analysis

Statistical data were expressed as mean ± standard error (mean ± S.E.M.). The statistical significance test between groups was analyzed through Student's t-test, and the continuous viability of nematodes was analyzed using the log-rank test analysis method. The p value was considered significant when *p<0.05, **p<0.01, and ***p<0.001.

2. Results

- DPPH radical scavenging ability

DPPH radical scavenging activity measurement-DPPH radical scavenging activity analysis was performed to evaluate the DPPH radical scavenging activity of the samples used in the experiment. The DPPH radical scavenging effect (IC50 value, 239.3 μg/ml) of the beet extract was not better than that of vitamin C (IC50 value, 19.81 μg/ml) as shown in FIG. 1, but the free radical DPPH scavenging effect was good. The mean value was obtained from three experiments.

- Superoxide inhibitory activity

Superoxide inhibitory activity was performed to evaluate the superoxide inhibitory activity of the sample used by measuring the photochemical activity using an evaluation system composed of methionine, riboflavin, and NBT. In the beet ethanol extract, the superoxide inhibitory effect was almost similar to that of the beet ethanol extract (IC50 value, 317.8 μg/ml) and vitamin C (IC50 value, 255.7 μg/ml), as shown in FIG. 2 . . The mean value was obtained from three experiments.

- Efficacy of increasing antioxidant enzyme (SOD, Catalase) activity

As a result of measuring the activity of SOD by using superoxide anion generated during the enzymatic reaction of xanthine oxidase with xanthine as a substrate, as shown in Fig. 3A, the group administered with the beet ethanol extract of C. and 250 and 500 μg/mL of beet ethanol extract increased SOD activity by about 6.2% and 14.4% (*p<0.05), respectively, compared to the control group. The activity of catalase that metabolizes hydrogen peroxide, an active oxygen species with strong reactivity, in the body, as shown in FIG. 3B, showed that the beetroot extract 250 and 500 μg/mL administered group showed a catalase activity of about 10% (*p<0.05), compared to the control group. It increased by 21.7% (***p<0.001) (FIG. 3).

- Reduction of reactive oxygen species (ROS)

In order to investigate the effect of reducing intracellular reactive oxygen species by concentration of the beet ethanol extract, H2DCF-DA was reacted with reactive oxygen species inside the nematode and fluorescence was observed. The decrease in reactive oxygen species fluorescence was 9.1% (*p<0.05) in the 250 μg/mL beet extract group and about 20.8% (**p<0.01) in the 500 μg/mL group on average compared to the control group. It was confirmed to decrease (Fig. 4).

- Efficacy of increasing expression of SOD-3 in transgenic nematodes

In order to check whether the protein to resist oxidative stress in the nematode increases, it was checked whether the SOD-expressing gene was increased. As a result of the experiment using the transgenic nematode CF1553 including SOD-3, the CF1553 transgenic nematodes treated with 250 and 500 μg/mL of beet ethanol extract showed a higher SOD-3::GFP expression rate (8.9%) compared to untreated nematodes. , 18.5%, *p<0.05, **p<0.01) (FIGS. 5A, 5B).

- Efficacy of increasing oxidative stress resistance

The effect of beet ethanol extract on the survival rate of nematodes under oxidative stress conditions was confirmed. In order to induce oxidative stress in nematodes, the maximum survival time of control nematodes cultured in a 96-well plate containing 1 mM juglone M9 buffer was 21 hours, but with 250 μg/mL of beet ethanol extract, the survival time was increased to 22 hours. and the survival time was increased to 26 hours at a concentration of 500 μg/mL. The average survival time of the control group was 13.7±0.9 hours, but 15.8±0.9 hours in the 250μg/mL treatment group and 18.1±1.2 hours in the 500 μg/mL concentration group, which improved the survival time by 15.8% and 32.2%. (*p<0.05, **p<0.01) (Fig. 6 A, B).

Antioxidant activity was shown in DPPH radical scavenging test and superoxide scavenging activity test of beet ethanol extract. As a result of measuring the viability of nematodes by applying oxidative stress to determine the factors affecting the lifespan extension of the beetroot extract using C. elegans, the survival rate was increased at 500 μg/mL of the beetroot extract. These results suggest that beetroot ethanol extract has development value as a resource for the development of substances related to antioxidants.

<Anti-inflammatory experiment>

1. Materials and test methods

- Extraction and concentration

Red beet raw material (about 20 kg) was dried in the shade and then 2 kg were extracted with ethanol while shaking with ethanol twice at 50°C for 5 hours. The extract was concentrated under reduced pressure in a water bath to obtain ethanol extract, which was stored in a freezer (-200) and used as a sample.

- Measurement of NO production inhibitory action

The concentration of NO was measured using the Griess Reagent System according to the method of Wang et al. 1) for the concentration of nitrite in the culture medium. Raw 264.7 cells were aliquoted in a 96-well plate to 5.0 × 105 cells/well and incubated for 18 hours, then transferred to extracts of red beet and 6 other types, betalain, DL-sulforaphane, formula 10, 100, and 1000 μg/mL. After 1 hour of treatment, 10 μg/mL of LPS was treated and cultured for 24 hours. The same amount of Griess Reagent as the culture medium was added and reacted at room temperature for 10 minutes, and then absorbance was measured at 540 nm. The NO concentration of the culture medium was determined using a standard curve for each concentration of sodium nitrite.

- Measurement of proinflammatory cytokines - TNF-α, IL-1β, IL-8

RAW264.7 cells were aliquoted to a 96-well plate at 1.0 × 105 cells/well and cultured for 18 hours. Extracts of beet and 6 other types, betalain, DL-sulforaphane, and total extract at concentrations of 10, 100, and 1000 μg/mL were used. After pre-treatment and 1 hour after treatment with LPS 10 μg/mL, incubated for 24 hours. After obtaining a cell culture medium, TNF-α, IL-1β, and IL-8 contained in the culture medium were measured using an ELISA kit.

- Statistical analysis

Experimental results were calculated as mean ± standard error (Mean ± S.E.), and the students' t-test was used to verify the significance between each group. When p<0.05, it was considered to be significant.

2. Results

- Measurement result of NO production inhibitory action

Macrophages play a central role in the inflammatory response, and reactive nitrogen species (RNS) such as NO, HNO2, and ONOO- are produced in large amounts due to the immune response of macrophages, neutrophils and other immune cells during the inflammatory response. NO, a type of reactive oxygen species that has recently been known to play an important role in inducing inflammation, is a highly reactive biogenic molecule, produced from L-arginine by NOS (nitric oxide synthase). It is involved in inflammatory cytokines such as TNF-α and LPS, and is expressed when stimulated.

As a result of the inhibition of NO production in RAW 264.7 cells using LPS used as an inflammatory substance, red beet was very good. (Fig. 7)

In order to confirm this inhibitory effect, it was compared with gold, which is a positive control. When comparing beets at 1000mg/ml concentration, when LPS-induced inflammation was 100%, beets were able to suppress the inflammation expression by 54.68% and 45.32%. (Fig. 8) As a result, it was confirmed that beets have an excellent NO production inhibitory effect.

- Proinflammatory cytokine measurement results (each result is shown in FIGS. 9 to 14)

In terms of TNF-a production inhibitory power, beets showed 75.05% and IL-1β production inhibitory effects were 41.1% when LPS value was 100% at 1000 mg/ml concentration, and gold showed 43.5% as a positive control. As a result of the study on the IL-8β production inhibitory effect, when the LPS value was 100% at the concentration of 1000 mg / ml, the beetroot extract showed 40.7% and the positive control group, gold, showed 43.0%.

<Lung cancer treatment effect>

Selection of primary ingredients: Beetroot, Celeriac, Vitelotte, Burdock, Scutellaria baicalensis, Cabbage, Brussels Sprouts, Broccoli , Potato, Radish, and Curcuma longa 12 kinds of raw materials were excluded through the preliminary screening test for anticancer growth.

Dulbeco's modified eagle medium (DMEM) was used as the cancer cell culture medium, and GIBCO (Grand Island Biological Co., NY, USA) products were used for fetal bovine serum (FBS), antibiotics, and trypsin-EDTA. Methanol (MeOH) and ethanol (EtOH) as organic solvents for solvents are from Duksan Pharmaceutical (KOREA), and other reagents from Sigma-Aldrich Co., Ltd. Ltd (Irvine, UK) reagent was used. My CO2 (Science Technology, Korea) was used for the CO2 incubator, and a ZEISS inverted microscope (Axiovert s 100, Germany) was used for the cell microscope.

Eight cell lines were used in the study: A549 (lung cancer), DU-145 (prostate cancer), HeLa (uterine cancer), MCF-7 (breast cancer), SNU-182 (liver cancer), SNU-1196 (biliary tract cancer), and HepG2. (liver cancer) and SNU -1 (stomach cancer). All of them used human-derived cancer cells, and they were purchased from the Korea Cell Line Bank (KCLB). Of these, two types of HepG2 (liver cancer) and SNU-1 (gastric cancer) were excluded from the study due to culture and proliferation problems, and a total of 6 types were studied.

- Extraction and concentration

Beetroot, Celeriac, Vitelotte, Burdock, Scutellaria baicalensis, Brussels Sprouts , Broccoli raw materials (approximately 20 kg) were minced in the shade and then extracted twice with warm needles at 50 °C for 5 hours while shaking 2 kg with methanol. The extract was concentrated under reduced pressure on a water bath to obtain methanol extract, which was stored in a freezer (-200) and used as a sample.

cell culture

In a 100 mm petri dish, DMEM (containing 5% FBS) medium and about 2 × 104 cells/5 ml were incubated in a CO2 incubator for 48 hours, and a suspension of each cell was prepared and tested by the cck-8 method.

Cytotoxicity test (cck-8)

After dispensing 100 μl of cancer cell suspension (5×103 cells/well) in a 96-well plate,

Pre-incubate the plate at 37 °C and 5% CO2 conditions for 24 hr. 10 μl of samples such as beet extracts of various concentrations are added to the 96-well plate. After incubation for 24 hours under the same conditions, 10 μl of CCK-8 solution was added to each well of the plate.

Incubate the plate for 2 h and measure the absorbance at 450 nm using a microplate reader. Since the absorbance value is proportional to the number of living cells, the inhibition of cancer cell growth of each sample can be known as the absorbance value.

me. Experiment result

(1) Growth inhibitory effect of each raw material on lung cancer cell A549

As a result of the study on the cancer cell proliferation inhibitory effect of 10 extracts against lung cancer cell A549, beets were the best, and when the Coctrol value was 100% at 1 mg/ml concentration, beets grew only by 0.167, showing an inhibitory power of 98.4%. As a positive control, gold 99.4%, Celeriac 91.8%, Purple potato (Vitelotte), 62.8% Beetroot 1mg, 98.4%, Celeriac 91.8%, Gold (Scutellaria baicalensis), 99.4% showed

Claims (3)

A composition for treating lung cancer comprising a beet extract as an active ingredient. The method of claim 1,
The beet extract is a composition for treating lung cancer, characterized in that it has the ability to inhibit riboflavin-derived free radicals. The method of claim 1,
The beet extract is a composition for the treatment of lung cancer, characterized in that it has the ability to inhibit NO production.

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