75 results about "Breast cancer stem cell" patented technology

Human breast tumors contain hetrogeneous cancer cells. using an animal xenograft model in which human breast cancer cells were grown in immunocompromised mice we found that only a small minority of breast cancer cells had capacity to form new tumors. The ability to form new tumors was not a slochastic property, rather certain populations of cancer cells were depleted for the ability to form new tumors, while other populations were enriched for the ability to form new tumors. Tumorigenic cells could be distinguished from non-tumorigenic cancer cells based on surface marker expression. We prospectively identified and isolated the tumorigenic cells as CD4430CD24− / lowLINEAGE A few as 100 cells from this population were able to form tumors the animal xenograft model, while tens of thousands of cells from non-tumorigenic populations failed to form tumors. The tumorigenic cells could be serially passaged, each time generating new tumors containing and expanded numbers of CD44+CD24 Lineage tumorigenic cells as well as phenotypically mixed populations of non-tumorigenic cancer cells. This is reminiscent of the ability of normal stem cells to self-renew and differentiate. The expression of potential therapeutic targets also differed between the tumorigenic and non-tumorigenic populations. Notch activation promoted the survival of the tumorigenic cells, and a blocking antibody against Notch 4 induced tumorigenic breast cancer cells to undergo apoptosis.

The invention provides a preparation method of a double chimeric antigen receptor gene modified T lymphocyte targeting breast cancer stem cells. The preparation method is characterized in that integrin-associated protein (CD47) and transcriptional coactivator (TAZ) are both highly expressed in breast cancer tissues, especially in the breast cancer stem cells; by established CD47 and TAZ over-expressed breast cancer cells, higher proliferation and metastasis capacity and cancer stem cell features are shown. Extracellular domain of the two breast cancer stem cell antigens CD47 and TAZ are targeted to generate monoclonal strains in specific binding under immunity action, and single-chain antibodies in specific binding with the CD47 and TAZ by genetic recombination are obtained to construct a human CD47 and TAZ containing double chimeric antigen receptor gene recombined to a virus vector to transfect human T lymphocyte. By high expression of the double chimeric antigen receptor gene in specific binding with human CD47 and TAZ expressing breast cancer stem cells, a first signal and a costimulatory signal can be activated to trigger anti-breast-cancer cytotoxicity, and high cytotoxicity in in-vivo and in-vitro anti-cancer experiments is achieved.

The invention relates to the technical field of medicines, and provides a docetaxel and sulforaphane loaded hyaluronic acid modified poly(lactic-co-glycolic acid) copolymer self-assembled nano-particle, a preparation method thereof and an application of the self-assembled nano-particle in preparing a medicine for treating breast cancer. The microscopic structure of the nano-particle comprises a hydrophobic PLGA (poly(lactic-co-glycolic acid)) core and a hydrophilic HA (hyaluronic acid) shell, wherein the PLGA core is used for wrapping a medicine DTX (docetaxel) or SFN (sulforaphane), and the hydrophilic shell HA can target a breast cancer stem cell highly expressing a CD44 receptor. In-vitro cytotoxicity tests indicate that the drug-loaded nano-particle can play a good role in resisting differentiation of mammary gland cells and breast cancer stem cells when compared with free medicines; in-vivo tumor inhibition experiments indicate that the drug-loaded medicine is more effective than free medicines and combined free medicines, and has a small systematic toxicity. The nano-particle can play a role in simultaneously performing target differentiating on mammary gland cells and breast cancer stem cells, and a new strategy is provided to treatment of breast cancer.

MicroRNA markers of breast cancer stem cells (BCSC) are provided herein. The markers are polynucleotides that are differentially expressed in BCSC as compared to normal counterpart cells. Uses of the markers include use as targets for therapeutic intervention; as targets for drug development, and for diagnostic or prognostic methods relating to breast cancer and BCSC cell populations. BCSCs have the phenotype of having lower expression of certain miRNAs compared to normal breast epithelial cells, or to cancer cells that are not cancer stem cells.

Immunogenic compositions, cancer vaccines and methods for treating cancer are provided. Compositions comprising: (a) a glycan such as Globo H or an immunogenic fragment thereof, wherein the glycan is conjugated with a carrier protein by a linker such as para-nitrophenyl; and (b) an adjuvant comprising glycolipid capable of binding CDId on a dendritic cell, such as an a-galactosyl-ceramide derivative, wherein the immunogenic composition induces an immune response that induces a higher relative level of IgG isotype antibodies as compared to IgM isotype antibodies, are provided. Immunogenic compositions comprising the carrier protein diphtheria toxin cross-reacting material 197 (DT-CRM 197) and the adjuvant C34 are provided. Antibodies generated by immunogenic compositions disclosed herein further neutralize at least one of the antigens Globo H, stage-specific embryonic antigen-3 (SSEA-3) and stage-specific embryonic antigen-4 (SSEA-4). Therapeutics against breast cancer stem cells comprising immunogenic compositions comprising Globo H, SSEA-3 or SSEA-4 conjugated with DT-CRM 197.

This invention relates to a method for separating breast cancer stem cells. The method comprises: digesting breast cancer cells into single cell suspension, preparing Percoll discontinuous density gradient separating medium, mixing with single cell suspension, centrifuging to separate cell subsets, detecting by comparing the experimental tube and the control tube, adjusting the density of Percoll discontinuous density gradient separating medium until the content of the cell subsets related to the breast cancer stem cells reaches predetermined enrichment degree, and screening with a flow cytometry. The method utilizes Percoll screening or serum-free culture to enrich breast cancer stem cells, and then screens with a flow cytometry to ensure the screening efficiency. The ratio of the breast cancer stem cells after primary enrichment is higher than 25%, while that after low cytometry screening is higher than 99%.

The invention discloses a screening kit of malignant breast cancer stem cells. The screening kit comprises an optional related reagent for detecting CXCR6 gene expression. The invention also discloses application of the reagent for detecting the CXCR6 gene expression in preparation of a reagent for screening the malignant breast cancer stem cells. The kit provided by the invention is capable of accurately detecting whether a sample to be tested contains the malignant breast cancer stem cells, and provides a reliable index for diagnosis and treatment of clinical breast cancer, and also is bright in clinical application prospect.

The invention discloses a breast cancer molecular probe, which is composed of a signal component and an affinity component. The breast cancer molecular probe is characterized in that the affinity component is a component for specific combination of breast cancer stem cell targets; and the signal component is composed of a near-infrared fluorescent signal unit and a magnetic resonance signal unit. The preparation method comprises the following steps: coupling the signal function unit with monoclonal antibodies which are specific surface markers of breast cancer stem cells by adopting a chemical covalent coupling method; and purifying. Aiming at the molecular probe which is specifically combined with multiple targets of the breast cancer stem cell targets, in vivo detection and quantitative analysis of the breast cancer stem cells can be solved by virtue of the molecular probe; a basis is provided for a new strategy for imaging diagnosis and targeted treatment therapeutic effect evaluation of solid tumor stem cells; a 'new mode' is provided for early diagnosis and grading of tumors; precise information and an evaluation method are provided for tumor treatment; and a diagnostic information foundation is laid for further solving reappearing and transferring problems.

The invention provides a separation method of breast cancer stem cells and belongs to the field of cell separation. The separation method comprises the following steps of: (1) preparing a hydrothorax single-cell suspension; (2) carrying out immunomagnetic bead negative sorting, and separating and removing non-breast-cancer cells, so as to obtain breast cancer epithelial cell sap; and (3) carrying out enrichment and purification on the breast cancer epithelial cell sap obtained in the step (2) by utilizing a non-adhesive culture method, so as to obtain breast cancer stem cell spheroids. Compared with the prior art, the separation method has the beneficial effects that the surfaces of the cells obtained by utilizing the separation method are not interfered by antibodies and magnetic beads; the separation efficiency of the breast cancer cells and the purity of the breast cancer cells after the breast cancer stem cells are enriched can be guaranteed; and not only can a small quantity of breast cancer epithelial cells be effectively separated, but also the separation time is shortened, and the production cost is lowered, so that the separation method is safe and effective.

The invention discloses a three-dimensional culture method for screening breast cancer stem cells. According to the three-dimensional culture method, hydrogel is adopted for conducting three-dimensional culture on the human breast cancer cells to directly obtain breast cancer stem cell microspheres which are in ESA+CD44+ / CD24- / low phenotypes and obviously have the characteristics of tumor stem cells. The method is a reliable breast cancer stem cell screening method. The method is simple to operate and has the advantages that the costs of reagent consumption, instrument usage, human resources and the like are greatly lowered compared with the prior art such as a flow cytometer screening method.

The invention discloses a lovastatin silicon plastid of a targeted breast cancer stem cell and a preparation method thereof.The lovastatin silicon plastid is prepared from, by weight, lovastatin 2%-30%, organic-inorganic compound lipid 20%-70%, phosphatidyl ethanolamine-polyethylene glycol 2000 1%-20% and distearoyl phosphatidylcholine 10%-60%.The lovastatin silicon plastid overcomes the shortcoming that the bioavailability of lovastatin drugs in the prior art is low.

The invention discloses a method for separating and purifying breast cancer stem cells using an immunomagnetic bead sorting system. The steps include: firstly preparing a single cell suspension, and secondly performing negative sorting by immunomagnetic beads to separate and remove normal cells in breast cancer cells , and finally perform immunomagnetic bead positive sorting to purify breast cancer stem cells. The method is simple and quick to operate, does not require expensive experimental equipment, has a high survival rate of cells after screening, and has high CD44++CD24- / + specificity, and will have broad application prospects in breast cancer stem cell research.

The invention discloses a breast cancer stem cell cryopreservation protection agent and a prepared cryopreservation protection kit. The cryopreservation protection agent consists of serum-free medium freeze-dried powder and fetal calf serum FBS, wherein the serum-free medium freeze-dried powder is prepared by the following steps: adding conventional growth factors, galanthamine and lycorine into a DMEM / F12 culture solution to obtain a culture solution of which the EGF concentration is 15-25g / L, the BSA concentration is 3-5g / L, the insulin concentration is 4-6mg / L, the galanthamine concentration is 50-100nmol / L and the lycorine concentration is 50-100nmol / L, wherein the conventional growth factors include recombinant human epidermal growth factor EGF, bovine serum albumin BSA, insulin and B27; storing in a shaking table for 20-28h at 37 DEG C in the presence of 5% CO2; and performing freeze drying. The cryopreservation protection agent disclosed by the invention can effectively reduce the damage of breast cancer stem cells caused by cryopreservation, the recovered cells have a high survival rate and strong tumorigenic ability and are not much different from those before cryopreservation, and thus the invention is remarkably superior to the prior art.

Disclosed is a method for inhibiting the growth of breast carcinoma stem cells. that express High Molecular Weight -Melanoma Associated Antigen (HMW-MAA). The method comprises administering to an individual a composition comprising an antibody reactive with HMW-MAA or a fragment of such an antibody in an amount effective to inhibit the growth of the breast carcinoma cells. Also provided are methods for inhibiting metastasis of breast carcinomas and methods for identifying HMW-MAA+ breast cancer stem cells.

Human breast carcinoma SK-BR-3 cell strain as source cell for building train adapted by the invention is continuously inoculated on female NOD / SCID mouse mammary gland fat pad and filtered in pressure of low dose chemotherapy drug tail intravenous injection to build human breast carcinoma cell line SK-3rd rich in tumour breast carcinoma stem cell. The proper of knub stem cell in the cell is increased greatly comparing with non NOD / SCID vivo passage source cell. The formation rate of saccule and the number of cell in the saccule are increased greatly in case of cell suspension culture. The ratio of undifferentiation cancer cell is reduced but is higher evidently than source cell in process of SK-3rd saccule cell inducing differentiation. The saccule cell formed from SK-3rd cell has carcinoma stem cell phenotype which gradually dies out in process of differentiation. NOD / SCID mouse orthotopic transplantation has high tumour-forming rate, high long-distance transferring rate, short tumour-forming periodic time and less number of cells needed by tumour-forming. The invention is an ideal mould for studying breast carcinoma stem cell and has wide applications foreground for disclosing generation mechanism of breast carcinoma and neoplastic treat.

The invention discloses a micro-fluidic chip for breast cancer stem cell culture and pharmaceutical analysis, which is used for solving the technical problem of poor practicality of an existing micro-fluidic chip. According to the technical scheme, the micro-fluidic chip comprises four same array units, each array unit adopts a three-layer PDMS structure, namely a top layer, a medium layer and a bottom layer, and a polycarbonate film is arranged between each medium layer and the respective bottom layer; every two array units share a cell suspension inlet and a cell suspension outlet; a cell suspension inlet, a cell suspension waste liquid outlet and a cell suspension inlet passage are formed in each top layer; a cell culture cavity is formed in each medium layer; and each bottom layer is provided with a culture medium inlet, a drug inlet, a density gradient generator, a perfusion passage, a waste liquid passage and a waste liquid outlet. The micro-fluidic chip can be used for supplying more suspending space for suspension cells by constructing the cell culture cavity having a high aspect ratio, so that injury of fluid shearing force for the suspension cells can be reduced, and the aims of culturing beast stem cells and performing target pharmaceutical analysis can be achieved; and the practicality is good.

The invention discloses an electrochemical detection method for stem cells. The electrochemical detection method is characterized in that breast cancer stem cells are identified by polypeptide; the Nterminal of the polypeptide is modified with an azide group; an electrochemical signal is obtained by a copper-catalyzed azide-alkynyl Husigen cycloaddition reaction. According to the method providedby the invention, identification of the breast cancer stem cells is formed by designed and synthesized polypeptide and peptide fibers formed by self-assembly of the polypeptide, further silver nanoparticles with electrochemical response are introduced by means of click chemical reaction, and quantitative detection of target breast cancer stem cells is realized. The method has the advantages of high sensitivity and high specificity; no antibody needs to participate in, so the electrochemical detection method has a broad application prospect in the field of clinical diagnosis and the like.

The invention discloses a reagent and kit for enriching breast cancer stem cells from breast cancer cell strains. The reagent includes galantamine and lycorine as a screening agent. The breast cancer cell strains are MCF-7 breast cancer cell strains. The kit comprises a double-antibody, galantamine and lycorine as a screening agent, a screening-agent-dissolving auxiliary agent, sodium sulfite as an anti-oxidant, a recombinant human epidermal growth factor (EFG) which is a conventional growth factor, bovine serum albumin (BSA), insulin and B27. The provided reagent can enrich breast cancer stem cells from human breast cancer MCF-7 cell strains in a convenient, fast and efficient way, the cell spheroids are short in forming period, and the content of cancer cells and SP cells marked by CD44<+>CD24<- / low> molecules in microsphere cells is high. The developed kit can improve the efficiency for enriching cancer stem cells by science researchers.

The invention provides new application of a CD44 plasmid. The CD44 plasmid can be effectively used for enhancing the in-vitro damaging effect of DC-CIK on mammary cancer stem cells.

The present invention provides an application of a compound for targeted inhibition of CDK7 in preparation of inhibited triple negative breast cancer stem cells. The compound for targeted inhibition of CDK7 is THZ1. The CDK7 targeting inhibitor THZ1 can specifically inhibit growth and stemness maintenance of the triple negative breast cancer stem cells and in-vivo tumorigenesis and growth of the stem cells. Transcriptome sequencing and subsequent biochemical experiment verifications show that the CDK7-targeting inhibitor THZ1 can specifically regulate and control a tumor stemness Notch-Hes signaling pathway, promote stemness transformation of triple-negative breast cancer stem cell subsets, transform stemness cell subsets into non-stemness cells, cause cycle arrest of the triple-negative breast cancer stem cells, inhibit DNA damage repair, promote cell apoptosis, meanwhile, can also directly cause the apoptosis of the stem cells, can remarkably influence growth of the triple-negative breast cancer stem cells, and meanwhile, can play a synergistic sensibilization function in the radiotherapy of the triple-negative breast cancer stem cells.

The invention discloses an application of miRNA specific oligonucleotides in preparation of a kit for predicting hormone receptor positive breast cancer preoperative chemosensitivity. The kit is usedfor obtaining a risk score by detecting the expression level of at least one miRNA gene from a subject sample and combining a gene risk stratification calculation formula, whether the subject has therisk of hormone receptor positive breast cancer preoperative chemosensitivity or not is evaluated, a truncation value is selected as a judgment basis, the subject is at the high risk of preoperative chemosensitivity when the value is higher than the truncation value, and the subject is at the low risk of preoperative chemosensitivity when the value is lower than the truncation value; wherein the at least one miRNA gene is miR-200c. The invention further discloses a preparation method of the miRNA gene. According to the invention, the HR positive breast cancer preoperative chemosensitivity score is obtained by detecting the relative expression quantity of miRNA related to breast cancer stem cells according to a risk scoring model, so as to evaluate whether a patient is suitable for preoperative chemotherapy or not.

The invention discloses a tumor stem cell vaccine as well as a preparation method and application thereof. A breast cancer stem cell vaccine provided by the invention comprises inactivated breast cancer stem cells, mannatide and fat emulsion which are mixed according to the volume ratio of 3:1:1. The preparation method of the breast cancer stem cell vaccine comprises the following steps: obtaining breast cancer stem cells; inactivating the breast cancer stem cells; freeze-thawing the inactivated breast cancer stem cells into normal saline, and mixing the inactivated breast cancer stem cells with adjuvant mannatide and the fat emulsion, so as to obtain the breast cancer stem cell vaccine. The invention further provides the application of the breast cancer stem cell vaccine, prepared according to the preparation method provided by the invention, in preparing medicines for treating breast cancer. The invention provides a novel way for the treatment of breast cancer and the preparation of the vaccine, an active curative effect on a breast cancer patient is achieved, the immune function of the patient is enhanced, the lifetime of the patient is prolonged, and the active effect is achieved.

The invention provides a composition for detecting breast cancer stem cells. The composition comprises a first antibody-nucleic acid conjugate specifically binding to cell surface CD44 protein, a second antibody-nucleic acid conjugate specifically binding to cell surface CD24 protein, a DNA single-chain T chain and a DNA single-chain F chain, wherein the T chain and a nucleic acid chain of the first antibody-nucleic acid conjugate are complementarily bound, the F chain and a nucleic acid chain of the second antibody-nucleic acid conjugate are complementarily bound, a programmed DNA loop is formed, signal markers on the surfaces of the breast cancer cells with CD44 positive and CD24 positive phenotypes are erased, and signal probes marked to the breast cancer stem cells are retained. The composition provided by the invention realizes identification and effective distinguishing of the breast cancer stem cells and the breast cancer cells, and has the characteristics of simple and convenient operation, accurate result, low technical level requirement, small data analysis amount and the like.

The invention discloses two breast cancer stem cell lines capable of being stably cultured and keeping original characteristics, separation method and uses thereof. A breast cancer tumor specimen just cut off from the tumor is taken, then is subjected to collagenase digestion and is filtered; a flow cytometer sorts CD44<+>, CD24<-> and ALDH1<+> cells, the cells are adjusted to a purity of more than 98%; a stem cell culturing agent for breast cancer cell is prepared; the sorted cells are screened in a monoclonal manner and the obtained monoclone cells are inoculated in an ultralow adhesion culture dish containing breast cancer stem cell culturing agents; penicillin and streptomycin are added in the original culturing medium; culturing is performed in CO<2> cultivation box. In the continuous culturing process, the breast cancer stem cell lines capable of being stably cultured and keeping original characteristics keep ball formation ability, self-updating ability, CD44<+>, CD24<-> / low Lin- ALDH1<+> phenotype, pluripotency differentiation ability, in vitro soft agar tumorigenesis ability, in vivo tumorigenesis ability in nude mouse, and in vivo tumorigenesis pluripotency differentiation ability in nude mouse.

The invention relates to a preparing method of breast cancer specificity CTLs, and belongs to the technical field of biological pharmacy. The preparing method includes the steps that breast cancer stem cells are separated from autologous cancer tissue of a breast cancer patient, then the cell fusion technology is used for fusing autologous DC cells of the patient and the autologous breast cancer stem cells to obtain fused cells with characteristics of the breast cancer stem cells and antigen transmissibility of the DC cells, and then cytotoxic T-cells (CTLs) capable of specifically killing the breast cancer stem cells can be generated in quantity through stimulation. The obtained CTLs capable of specifically killing the breast cancer stem cells are returned into the body of the patient, and the effects of killing the breast cancer stem cells and preventing relapse and transfer can be achieved.

The invention discloses a preparation method and an application of a BCSC (breast cancer stem cell) antigen. The method comprises the following steps: culturing a BCSC, adding hypochlorous acid to a solution for culturing the BCSC for uniform mixing, inducing apoptosis of the BCSC, cracking the BCSC treated by hypochlorous acid, and collecting a cell lysate to obtain the antigen, wherein hypochlorous acid added to the solution for culturing the BCSC has concentration of 20-80 mu M and treatment time of hypochlorous acid is 30-60 min. The antigen prepared with the method can be used for preparing tumor DC vaccine. The method for collecting the antigen after treating the BCSC with hypochlorous acid is not only economical and simple, but also can more strongly activate immune response of antigen-specific T cells and improve tumor killing capacity of the T cells.

The present invention provides a method for inhibiting the growth of cancer stem cells, particularly colorectal cancer stem cells, liver cancer stem cells, lung cancer stem cells or breast cancer stem cells, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of antimycin A or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.