CN112315973B

CN112315973B - Application of oyster mushroom polysaccharide selenoside-III anticancer active ingredient in preparation of medicine for resisting colon cancer

Info

Publication number: CN112315973B
Application number: CN202011251224.1A
Authority: CN
Inventors: 钟世安; 王嘉晖; 刘慧�; 张云山; 张卓敏; 沈剑
Original assignee: Hunan Wanzhen Biotechnology Co ltd
Current assignee: Hunan Wanzhen Biotechnology Co ltd
Priority date: 2020-02-17
Filing date: 2020-11-11
Publication date: 2023-09-26
Anticipated expiration: 2040-11-11
Also published as: CN112315973A

Abstract

The invention belongs to the technical field of anticancer drug development, and particularly discloses oyster mushroom polysaccharide selenoside-III with specificity and colon cancer resistance; the invention also discloses an obtaining method of the oyster mushroom polysaccharide selenoside-III and application of the oyster mushroom polysaccharide selenoside-III in preparation of colon cancer drugs. The research of the invention discovers that the oyster mushroom polysaccharide selenoside-III can be used as an anticancer drug to specifically inhibit the growth of colon cancer cells without damaging normal cells: can reduce toxic and side effects while ensuring drug effect.

Description

Application of oyster mushroom polysaccharide selenoside-III anticancer active ingredient in preparation of medicine for resisting colon cancer

Technical Field

The invention relates to the technical field of medicines, in particular to an oyster mushroom polysaccharide selenoside-III anticancer active ingredient with high anticancer effect and low toxic and side effect.

Technical Field

Colon cancer is one of the most common malignant tumors of the digestive system today, and is also one of the common tumors of the clinical oncology. The incidence rate of the traditional Chinese medicine is always in an ascending trend, the traditional Chinese medicine gradually gets younger, the incidence rate of men is higher than that of women, and the incidence rate of urban areas is higher than that of rural areas. The colon cancer has no specific clinical symptoms in early stage, often enters middle and late stages when hematochezia occurs, and is often confused with hemorrhoids or other gastrointestinal inflammation symptoms, so that the treatment of the colon cancer can be greatly delayed. Surgical treatment is performed in advanced stages of colon cancer patients, with survival rates of only 10%. At present, the prevalence rate of colon cancer is high, and research on anticancer drugs which have specific killing ability to colon cancer and have no toxic or side effect on normal cells or tissues is not slow.

Disclosure of Invention

The invention aims to provide an oyster mushroom polysaccharide selenoside-III anticancer active ingredient (also called polysaccharide selenoside-III) with high anticancer effect and low toxic and side effects and application thereof in preparing anti-colon cancer drugs.

The second aim of the invention is to provide an anti-colon cancer drug containing the oyster mushroom polysaccharide selenoside-III anti-cancer active ingredient.

The prior art discloses extraction modes of oyster mushroom polysaccharide selenosides and an antioxidant effect, but the prior art rarely relates to the antioxidant effect of oyster mushroom polysaccharide selenosides. In fact, the present inventors have found that crude oyster mushroom polysaccharide (a mixture of various active ingredients) extracted from oyster mushrooms has substantially no anticancer activity. However, in the background of research in which crude oyster mushroom polysaccharide is known to have no anticancer activity, there is no motivation in the industry to screen out anticancer components therefrom, however, the present inventors have unexpectedly found that the oyster mushroom polysaccharide selenoside-iii isolated from the crude oyster mushroom polysaccharide has unexpectedly good activity against colon cancer, and at the same time has no toxic or side effects on normal cells, through intensive studies, so the following technical scheme is provided:

an application of oyster mushroom polysaccharide selenoside-III anticancer active component in preparing medicine for resisting colon cancer.

The oyster mushroom polysaccharide selenoside-III is a polysaccharide compound modified with organic selenium, and the polysaccharide compound is obtained by connecting monosaccharide units in glucose, galactose, mannose, glucuronic acid, galacturonic acid, xylose, glucosamine, fucose, arabinose, ribose and rhamnose through glycosidic bonds;

the content of glucose in the hydrolyzed polysaccharide compound is more than 80%, and the content of galactose, mannose and glucuronic acid is more than 1%; the content of each of galacturonic acid, xylose, glucosamine, fucose, arabinose, ribose and rhamnose is not higher than 1%;

preferably, the content of glucose in the hydrolyzed heteropolysaccharide compound is 82-84%, the content of galactose is 3-5%, the content of mannose is 2.5-3.5%, and the content of glucuronic acid is 1-2%; the content of each of galacturonic acid, xylose, glucosamine, fucose, arabinose, ribose and rhamnose is not higher than 1%.

The research of the invention discovers that the oyster mushroom polysaccharide selenoside-III has excellent anticancer activity, and moreover, the oyster mushroom polysaccharide selenoside-III can not damage normal cells while killing cancer cells.

For the anticancer drug, the normal cells are not damaged while inhibiting and killing cancer cells. However, the inventor further conducts intensive research on oyster mushroom polysaccharide selenoside-III, and discovers that the oyster mushroom polysaccharide selenoside-III not only has good anticancer activity, but also can solve the problem that the conventional anticancer active ingredients damage normal cells.

The monosaccharide composition analysis shows that the glucose content of the oyster mushroom polysaccharide selenoside-III after hydrolysis is more than 80%, the galactose, mannose and glucuronic acid content is more than 1%, and the monosaccharide composition is obviously different from the polysaccharide or polysaccharide derivative of the existing oyster mushroom source, so that the oyster mushroom polysaccharide selenoside-III is a newly obtained substance.

Preferably, the molecular weight of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient is 13000-18000.

Preferably, the organic selenium is modified in the polysaccharide compound by se=o and Se-C-O;

preferably, the selenium content is 24-26 mug/g.

The application of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient is characterized in that: the preparation process of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient comprises the following steps:

separating the crude polysaccharide by a cellulose exchange column, wherein the separation process comprises the following steps of sequentially carrying out distilled water elution, 0.05-0.15 mol/L sodium chloride solution elution, 0.25-0.35 mol/L sodium chloride solution elution and 0.45-0.55 mol/L sodium chloride solution elution;

collecting the eluent of 0.25-0.35 mol/L sodium chloride solution as target eluent, desalting, concentrating, and purifying by a sephadex column.

According to the preparation method, the oyster mushroom polysaccharide selenoside-III anticancer active ingredient with excellent anticancer activity and normal cell activity is not damaged can be unexpectedly obtained through the special elution mechanism.

The degreasing process of the alcohol aqueous solution comprises the following conditions: the alcohol-water solution is ethanol water solution, and the volume fraction of the ethanol is 75-85%; reflux degreasing;

degreasing an alcohol aqueous solution, performing solid-liquid separation, and dispersing and extracting the precipitate in water; the temperature of the water extraction is 80-90 ℃;

extracting with water to obtain extractive solution, concentrating, adding ethanol, precipitating with ethanol, and separating solid from liquid to obtain ethanol precipitate;

deproteinizing the alcohol precipitate to obtain crude polysaccharide.

Preferably, the cellulose exchange column is a DEAE-52 cellulose ion exchange column.

Preferably, the target eluent is desalted by dialysis means.

Preferably, the sephadex column is a G-100 sephadex column.

Preferably, the application uses the oyster mushroom polysaccharide selenoside-III anticancer active ingredient to prepare an anti-colon cancer drug capable of specifically apoptosis of HCT116 cells.

Further preferably, the use of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient for preparing a polypeptide which specifically causes HCT116 cell apoptosis and does not damage normal colon cells.

The oyster mushroom polysaccharide selenoside-III prepared by the invention can effectively kill cancer cells without damaging normal cell functions, and has important significance for the research of the oyster mushroom polysaccharide serving as an anticancer active ingredient in the development of the anticancer field.

The normal cells are preferably normal colon cells NCM460.

For example, when the concentration of the polysaccharide selenoside-III anticancer active ingredient is 600 mug/mL, the survival rate of cancer cells is about 60%, and the tumor inhibition rate is about 40%. However, at the same administration concentration, the survival rate of normal colon cells can reach more than 90 percent. So that the colon cancer cells can be inhibited and normal cells can not be damaged obviously.

The invention also provides an anti-colon cancer drug which specifically promotes HCT116 cell apoptosis and does not damage normal colon cells, and the drug comprises a pharmaceutically effective amount of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient.

Preferably, the medicine for resisting colon cancer, which specifically promotes apoptosis of HCT116 cells and does not damage normal colon cells, further comprises auxiliary materials for preparing the oyster mushroom polysaccharide selenoside-III anticancer active ingredient into any pharmaceutically acceptable dosage form.

Any pharmaceutically acceptable dosage forms are, for example, powder injection, capsules, dripping pills, medicinal granules and the like.

The anticancer activity of the polysaccharide selenoside-III anticancer active ingredient can be quantitatively analyzed by a cck-8 method, and can also be visually analyzed by an AO/EB fluorescent staining method. Acridine Orange (AO) is a selective fluorescent cationic dye that can permeate cell membranes and can stain both living and dead cells. Observing under a fluorescence microscope, wherein acridine orange can penetrate through a complete cell membrane, and the cell nucleus of a living cell is dyed into uniform green fluorescence; the apoptotic cells are condensed or broken into fragments with different sizes due to the chromatin, and the acridine orange dyes the cell nucleus into yellow green bright green fluorescence; dead cells fluoresce orange, but the fluorescence is weak and even vanishes. Ethidium Bromide (EB) can only stain cells with incomplete cell membranes, AO is used in combination with EB to stain necrotic cells orange or orange red, but this also includes cells that resemble viable nuclear morphology and do not have chromatin condensation. Therefore, AO/EB staining kit can distinguish normal cells, apoptotic cells and necrotic cells. The polysaccharide selenoside-III anticancer active ingredient can be observed to specifically cause HCT116 cells to apoptosis by an AO/EB staining method, and has no obvious damage to normal cells.

Advantageous effects

1. The invention surprisingly discovers that the oyster mushroom polysaccharide selenoside-III not only has good anticancer activity, but also can solve the problem that the existing anticancer active ingredients hurt normal cells.

2. By the preparation method, the oyster mushroom polysaccharide selenoside-III with good anticancer activity and no obvious toxic or side effect on normal cells can be obtained.

Drawings

FIG. 1 is an infrared spectrum (758 cm) of a component of oyster mushroom polysaccharide selenoside-III ^-1 The peak at the position is Se=O characteristic peak, 660cm ^-1 The peak of (C) is Se-C-O bond

FIG. 2 is a Gel Permeation Chromatography (GPC) diagram of oyster mushroom polysaccharide selenoside-III component

FIG. 3 shows the elution curves of the fiber columns of crude selenium polysaccharide of oyster mushroom (three components eluted from aqueous sodium chloride solution with concentration of 0,0.1 and 0.3mol/L respectively, named as selenoside I, component II and component III respectively)

FIG. 4 shows the selenium content (ICP) of the oyster mushroom polysaccharide selenoside-III component

FIG. 5 elution profile (HPLC) of oyster mushroom polysaccharide selenoside-III hydrolysate and reference monosaccharides

FIG. 6A is a Transmission Electron Microscope (TEM) image of oyster mushroom polysaccharide selenoside-III

FIG. 7 shows survival of colon cancer cells (HCT 116) after incubation of oyster mushroom polysaccharide selenoside-III fraction

FIG. 8 is a fluorescent image of colon cancer cells (HCT 116) after AO/EB staining after incubation of the oyster mushroom polysaccharide selenoside-III fraction. (pictures were taken with high content, 40×)

FIG. 9 is a flow chart of colon cancer cells (HCT 116) after staining with annexin V-FITC/PI apoptosis detection kit following incubation of polysaccharide selenoside-III

FIG. 10 is a scratch assay of colon cancer cells (HCT 116) after incubation with polysaccharose selenoside-III

FIG. 11 survival of colon cancer cells (HCT 116) after incubation of crude oyster mushroom polysaccharide

FIG. 12 survival of normal colon cells (NCM 460) after incubation of oyster mushroom polysaccharide selenoside-III fraction

FIG. 13 is a fluorescent image of normal colon cells (NCM 460) after AO/EB staining after incubation of the oyster mushroom polysaccharide selenoside-III fraction. (pictures were taken with high content, 40×)

FIG. 14 is a flow chart of normal colon cells (NCM 460) after staining with annexin V-FITC/PI apoptosis detection kit following incubation of polysaccharide selenoside-III

FIG. 15 survival of normal colon cells (NCM 460) after incubation of crude oyster mushroom polysaccharide

Detailed Description

The following examples are intended to illustrate the invention and are not intended to be limiting.

Example 1

Preparation of crude selenium polysaccharide

15g of selenium-enriched oyster mushroom powder (supplied by Hunan Wanzhen biotechnology Co., ltd.) is taken in a round-bottomed flask, 80% ethanol-water solution 120mL is added, stirring reflux is carried out for 3h at 65 ℃, suction filtration is carried out, a filter cake is obtained, and drying is carried out.

Water extraction: 10g of filter cake was taken in a flask, 300mL of ultrapure water was added thereto, stirred at 85℃for 3 hours, centrifuged, and the supernatant was collected and concentrated to obtain a viscous liquid.

Alcohol precipitation: dripping 4 times of absolute ethyl alcohol into the viscous liquid, standing at 4 ℃ for 24 hours, and centrifuging to obtain a precipitate.

Deproteinization: dissolving the precipitate in 100mL of water, adding sevage reagent with quarter volume, centrifuging to obtain supernatant, repeating for 6 times, dialyzing with dialysis bag, and lyophilizing to obtain crude polysaccharide (also called as crude Pleurotus ostreatus polysaccharide or crude selenium polysaccharide).

(II) refining selenium polysaccharide

(2.1) ion exchange column purification procedure: purifying polysaccharide by using DEAE-52 cellulose ion exchange column, dissolving 300mg of sample (crude oyster mushroom polysaccharide in step (I)) in 10mL of water, and loading by wet method. Then sequentially eluting with distilled water, 0.1,0.3 and 0.5mol/L sodium chloride solution in a gradient way. The polysaccharide content was checked by phenol-sulfuric acid method at 490nm in a tube-separating manner, and the elution profile was drawn and the main component was collected. Three components were separated in total, and the present invention was derived from a component eluted from a 0.3mol/L aqueous sodium chloride solution, namely, component 3.

(2.2) dialysis: the sample-water solution (component 3 of step (2.1)) was concentrated to 10-15mL using a rotary evaporator, then pH was adjusted to 7 with hydrochloric acid solution, dialyzed against deionized water for 36h, and then concentrated to 5mL.

(2.3) dextran column purification procedure: selecting G-100 glucan to purify component 3 (component in step (2.2)), loading 5mL of sample by a wet method, eluting with deionized water, detecting by a phenol-sulfuric acid method in a separation tube, concentrating, and freeze-drying to obtain refined oyster mushroom polysaccharide selenoside-III (also called polysaccharide selenoside-III).

Hydrolysis of oyster mushroom polysaccharide selenoside-III and analysis of monosaccharide components: the sample was weighed precisely into a 5mL amp bottle, 2.0mL (2 mol/L) trifluoroacetic acid was added to a 5.0mL ampoule, the tube was sealed, and acidolysis was performed at 110℃for 8 hours. Taking out and volatilizing trifluoroacetic acid, adding 2.0ml of water for redissolving, and completing the hydrolysis. After that, 250. Mu.L of the hydrolyzed solution was precisely aspirated into a 5mL EP tube, 250. Mu.L (0.6 mol/L) of sodium hydroxide and 500. Mu.L (0.4 mol/L) of a methanol solution of 1-phenyl-3-methyl-5-pyrazolone were added, and reacted at 70℃for 1 hour. Cooling in cold water for 10min; add 500 μl (0.3 mol/L) hydrochloric acid to neutralize, add 1mL chloroform to vortex for 1min, centrifuge for 10min at 3000r/min, carefully take the supernatant, extract 3 times. Taking supernatant fluid and completing derivatization. The reference sample was a sugar mixture solution containing 50. Mu.g/mL of each of the reference sugars, and derivatization was performed under the same conditions. Finally, the sample and the reference are eluted by an Xtime C18 high performance liquid phase device, an elution curve is recorded, and a monosaccharide component can be calculated through the peak area ratio of the sample and the reference (the eluent is 0.05M pH=6.7 potassium dihydrogen phosphate solution-acetonitrile=83-17). The monosaccharide composition analysis results are shown in Table 1 and FIG. 5.

TABLE 1 content of polysaccharide selenoside-III constituent monosaccharides of oyster mushroom

The following anticancer and normal cytotoxicity studies were performed using the oyster mushroom polysaccharide selenoside-III obtained in example 1, specifically:

example 2

Experiments on the efficacy of polysaccharide selenoside-III (obtained in example 1) on colon cancer cells

When the HCT116 cell density reached 80-90%, the cells were allowed to round by pancreatin, after which the digestion was stopped by adding cell culture medium. The cell suspension was diluted appropriately and counted in a blood cell counting plate, and the 96-well plate was placed in a cell incubator for incubation for 24 hours at a density of 8000 cells per well (96-well plate).

The culture medium in the 96-well plate was then replaced with serum-free cell culture medium and incubated in a cell incubator for 24h.

The previous serum-free medium was replaced with cell culture media (0. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 400. Mu.g/mL, 600. Mu.g/mL) containing different concentrations of the polysaccharide selenoside-III, and incubated for 24h. And the cell-free and simple cell culture solution is used as a negative control group.

mu.L of cck-8 reagent was added to each well, and after 2 hours of co-incubation, the absorbance OD was measured at 450nm with a microplate reader to calculate the cell viability.

Cell viability= (OD _{Experimental group} -OD _{Negative control group} )/(OD _{Blank group} -OD _{Negative control group} )。

Plates (96 well plates) were plated at 5000 cells per well, and the 96 well plates were placed in a cell incubator for 24h incubation. The culture medium in the 96-well plate was then replaced with serum-free cell culture medium and incubated in a cell incubator for 24h. Cell culture media (0. Mu.g/mL, 300. Mu.g/mL, 600. Mu.g/mL) containing different concentrations of the polysaccharide selenoside-III were used instead of the previous serum-free medium and incubated for 24h. Wash with PBS and add 90 μl of PBS,5 μl of OA solution and 5 μl of EB solution, stain for 5min, then wash with PBS, observe under high content (AO excitation 488nm, emission 515nm, EB excitation 518nm, emission 605 nm).

Afterwards, apoptosis was quantified by flow cytometry at 5×10 per well ⁵ Is incubated for 24h. The medium was refreshed with cell culture media (0. Mu.g/mL, 300. Mu.g/mL, 600. Mu.g/mL) containing different concentrations of the polysaccharide selenoside-III drug and incubated for 24h. Cells were then collected by digestion with annexin V-FITC/PI apoptosis assay kit and detected on a flow cytometer.

Scratch experiments were used to verify the effect of drugs on cell migration capacity at 1x10 per well ⁶ Cells were inoculated for 24h for adherence, then streaked with 200. Mu.L of gun tips, then cultured in a drug-containing serum-free medium for 0, 12h,24h,48h, and then observed under a fluorescent inverted microscope.

After 24h incubation of HCT116 cells with polysaccharide selenoside-III, the cell viability was about 60% at a drug administration concentration of 600 μg/mL, i.e. about 40% tumor suppression (as shown in FIG. 7). This may suggest from the front that the polysaccharide selenoside-III may be effective in inhibiting colon cancer cells. As shown in fig. 8, from the AO/EB staining results, MGC803 cells incubated without adding the drug did not see a significant apoptosis signal, and with increasing administration concentration, EB channel fluorescence was significantly enhanced, and apoptotic cells were significantly increased, which more intuitively showed that polysaccharide selenoside-iii can effectively induce apoptosis of gastric cancer cells. As shown in FIG. 9, the proportion of apoptotic cells was 42.3% at a drug administration concentration of 600. Mu.g/mL. As shown in fig. 10, as the administration concentration increases, the ability of scratch recovery gradually decreases, and as a result, it was demonstrated that the polysaccharide selenoside-iii can inhibit the migration ability of tumor cells to some extent.

Comparative example 1:

crude oyster mushroom polysaccharide (without column purification, example 1 step (1) crude oyster mushroom polysaccharide) pharmacodynamic experiment on human colon cancer cells

The previous serum-free medium was replaced with cell culture media (0. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 400. Mu.g/mL, 600. Mu.g/mL) containing crude oyster mushroom polysaccharide at different concentrations, and incubated for 24h. And the cell-free and simple cell culture solution is used as a negative control group.

As shown in fig. 11, after the HCT116 cells were incubated with the crude oyster mushroom polysaccharide for 24 hours, the cell viability did not change significantly with increasing concentration of the drug administered, which proves that the crude oyster mushroom polysaccharide had no significant inhibitory effect on colon cancer cells to some extent.

Example 3

Drug efficacy experiment of polysaccharide selenoside-III (example 1) on Normal cells

When the normal colon cell density of the human reaches 80-90%, the human colon is digested by pancreatin until the cells become round, and then the digestion is stopped by adding cell culture solution. The cell suspension was diluted appropriately and counted in a blood cell counting plate, and the 96-well plate was placed in a cell incubator for incubation for 24 hours at a density of 8000 cells per well (96-well plate).

Scratch experiments were used to verify the effect of drugs on cell migration capacity at 1X10 per well ⁶ Cells were inoculated with the cell density of (C) for 24h for adherence, then streaked with 200. Mu.L of gun tips, then cultured with drug-containing serum-free medium for 0, 12h,24h,48h, then subjected to fluorescent inverted microscopyAnd (5) observing the mixture.

As shown in FIG. 12, when normal NCM460 cells are incubated with the polysaccharide selenoside-III, the cell survival rate can reach more than 90% at the administration concentration of 600 mug/mL, so that the polysaccharide selenoside-III can be obtained without obvious damage to normal colon cells. As shown in FIG. 13, with increasing dosing concentration, there was no significant change in AO/EB staining, indicating that the polysaccharide selenoside-III had no significant damage to normal colon cells. As shown in FIG. 14, the flow cytometry results show that the proportion of apoptotic cells is only 1% when the drug adding concentration is 600 mug/mL, so that the polysaccharide selenoside-III has no obvious toxic or side effect on normal cells. The data show that the polysaccharide selenoside-III has no obvious injury effect on normal liver cells.

Comparative example 2:

drug efficacy experiment of crude oyster mushroom polysaccharide (crude oyster mushroom polysaccharide obtained in step (I) of example 1) on normal cells without column purification

When the cell density of normal colon of human origin reaches 80-90%, the cells are digested by pancreatin until the cells become round, and then the cell culture solution is added to stop the digestion. The cell suspension was diluted appropriately and counted in a blood cell counting plate, and the 96-well plate was placed in a cell incubator for incubation for 24 hours at a density of 8000 cells per well (96-well plate).

As shown in fig. 15, after incubating HCT116 normal cells with crude oyster mushroom polysaccharide for 24 hours, the cell viability did not change significantly after incubation with the drug administration, which proves that the crude oyster mushroom polysaccharide has neither inhibitory nor proliferation-promoting effect on normal cells to some extent.

Claims

1. The application of oyster mushroom polysaccharide selenoside-III anticancer active ingredient in preparing medicine for resisting colon cancer is characterized in that: the oyster mushroom polysaccharide selenoside-III anticancer active ingredient is used for preparing an anti-colon cancer medicament which can specifically apoptosis HCT116 cells and does not damage normal colon cells; the normal colon cells are NCM460 cell line;

the oyster mushroom polysaccharide selenoside-III anticancer active ingredient is a heteropolysaccharide compound modified with organic selenium; the heteropolysaccharide compound is obtained by connecting monosaccharides including glucose, galactose, mannose, glucuronic acid, galacturonic acid, xylose, glucosamine, fucose, arabinose, ribose and rhamnose through glycosidic bonds;

wherein, in the hydrolyzed heteropolysaccharide compound, the glucose content is 83.74%, galactose 4.04%, mannose 2.80%, glucuronic acid 1.55%, galacturonic acid 0.54%, xylose 0.37%, glucosamine 0.28%, fucose 0.20%, arabinose 0.11%, ribose 0.03% and rhamnose 0.01%;

the molecular weight of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient is 15306;

the organic selenium is modified in the polysaccharide compound through Se=O and Se-C-O;

the selenium content is 25.9 mug/g;

the preparation process of the oyster mushroom polysaccharide selenoside-III anticancer active ingredient comprises the following steps:

degreasing selenium-enriched oyster mushroom powder by an alcohol aqueous solution, extracting with water, precipitating with ethanol, deproteinizing to obtain crude polysaccharide;

separating the crude polysaccharide by a cellulose exchange column, wherein the separation process comprises the following steps of distilled water elution, 0.1 mol/L sodium chloride solution elution, 0.3mol/L sodium chloride solution elution and 0.5mol/L sodium chloride solution elution which are sequentially carried out;

collecting 0.3mol/L sodium chloride solution as target eluent, desalting, concentrating, and purifying with sephadex column.

2. The use according to claim 1, wherein: the degreasing process of the alcohol aqueous solution comprises the following conditions: the alcohol-water solution is an ethanol water solution, and the volume fraction of ethanol is 75-85%; reflux degreasing;

deproteinizing the alcohol precipitate to obtain crude polysaccharide.

3. The use according to claim 1, wherein: the cellulose exchange column is a DEAE-52 cellulose ion exchange column.

4. The use according to claim 1, wherein: and desalting the target eluent by a dialysis means.

5. The use according to claim 1, wherein: the sephadex column is a G-100 sephadex column.