Use of Recombinant Humanized Collagen Type I in the Treatment of Breast Cancer

Abstract

The present invention belongs to the field of biomedicine, and specifically relates to the use of recombinant type I humanized collagen in the treatment of breast cancer. The present invention provides the use of recombinant type I humanized collagen in the preparation of a drug for preventing and / or treating breast diseases; wherein, the recombinant type I humanized collagen comprises n repeats of the sequence shown in SEQ ID No.1, n is an integer greater than or equal to 1, and when n is an integer greater than or equal to 2, the repeat sequences are directly connected; optionally, an amino acid sequence capable of being excised by TEV protease is included at the N-terminus of the recombinant type I humanized collagen polypeptide. The research results show that recombinant type I humanized collagen can significantly inhibit the growth and proliferation ability of breast cancer cells, inhibit the migration characteristics of tumor cells, and at the same time promote the dormancy of breast cancer cells.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the use of recombinant type I humanized collagen in the treatment of breast cancer. Background Art

[0002] Breast carcinoma (BC) is one of the most common malignant tumors in women and currently ranks first in the global cancer incidence. The treatment methods for breast cancer are limited, mainly including surgical resection, radiotherapy, chemotherapy, and hormone therapy, etc.

[0003] Triple negative breast cancer (TNBC) is a subtype of breast cancer, which refers to breast cancer with negative results in immunohistochemical examination of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). It has a high recurrence and metastasis rate, and due to the lack of effective treatment targets, the current treatment methods are relatively single, mainly chemotherapy. Therefore, more effective and safe treatment methods are urgently needed.

[0004] Collagen is the most abundant protein in animals, accounting for about 30% of the total protein, and plays an important role in physiological processes such as cell adhesion. Due to its good biocompatibility and biodegradation safety, collagen has been widely used in the medical field. Until now, the vast majority of collagens used in various studies are from animal tissues and skin extracts. Collagen extracted from animals has poor water solubility and weak processability, which directly limits the development of many potential uses. However, collagen produced by genetic engineering technology can effectively overcome the above disadvantages.

[0005] The Chinese patent application with the application number 202110968550.2 and the invention name "A recombinant type I humanized collagen polypeptide and its preparation method and use" records the recombinant type I humanized collagen studied and produced by the inventors of the present invention before. The recombinant type I humanized collagen has the characteristics of high water solubility and high biological activity. There has been no report on the connection between recombinant type I humanized collagen and the treatment of breast cancer. Summary of the Invention

[0006] Problems to be Solved by the Invention

[0007] The present invention intends to use the recombinant type I humanized collagen in the Chinese patent application with the application number 202110968550.2 as a raw material to evaluate its effectiveness in the treatment of breast diseases such as breast cancer. Specifically, the present invention intends to use the recombinant type I humanized collagen to inhibit the proliferation activity and / or migration ability of breast cancer cells and promote the dormancy of breast cancer cells, so as to achieve the effect of preventing and / or treating breast cancer.

[0008] Solutions for Solving the Problems

[0009] The present invention provides the use of recombinant type I humanized collagen in the preparation of a drug for preventing and / or treating breast diseases;

[0010] wherein, the recombinant type I humanized collagen comprises n repeats of the sequence shown in SEQ ID No.1, n is an integer greater than or equal to 1, and when n is an integer greater than or equal to 2, the repeat sequences are directly connected; optionally, the N-terminus of the recombinant type I humanized collagen polypeptide comprises an amino acid sequence that can be excised by TEV protease.

[0011] Furthermore, the amino acid sequence that can be excised by TEV protease comprises the sequence shown in SEQ ID No.2.

[0012] Preferably, the sequence shown in SEQ ID No.2 is directly connected to the sequence shown in SEQ ID No.1.

[0013] Furthermore, the recombinant type I humanized collagen comprises:

[0014] a) the amino acid sequence shown in SEQ ID No.3;

[0015] b) an amino acid sequence having more than 90% identity with the amino acid sequence of SEQ ID No.3 and retaining the activity of the amino acid sequence shown in SEQ ID No.3;

[0016] c) an amino acid sequence with 1 or more amino acid residues added, substituted, deleted or inserted in the amino acid sequence of SEQ ID No.3 and retaining the activity of the amino acid sequence of SEQ ID No.3; or

[0017] d) an amino acid sequence encoded by a nucleotide sequence that hybridizes with the polynucleotide sequence encoding the amino acid sequence of SEQ ID No.3 under stringent conditions, and the amino acid sequence retains the activity of the amino acid sequence of SEQ ID No.3, and the stringent conditions are medium stringent conditions, medium-high stringent conditions, high stringent conditions or very high stringent conditions.

[0018] Preferably, the recombinant type I humanized collagen comprises the amino acid sequence shown in SEQ ID No. 3.

[0019] Furthermore, the breast disease is breast cancer.

[0020] Furthermore, the breast cancer includes luminal A breast cancer, luminal B breast cancer, HER-2 overexpressing breast cancer, basal-like breast cancer, triple-negative breast cancer, and normal breast-like breast cancer.

[0021] Furthermore, the breast cancer is luminal A breast cancer, HER-2 overexpressing breast cancer, basal-like breast cancer, or triple-negative breast cancer.

[0022] Furthermore, the recombinant type I humanized collagen prevents and / or treats breast cancer by inhibiting the proliferation ability of breast cancer cells, inhibiting the migration ability of breast cancer cells, and / or promoting breast cancer cells to enter the dormant cycle.

[0023] Furthermore, the inhibition of the proliferation ability of breast cancer cells includes inhibiting the DNA replication activity of breast cancer cells.

[0024] Effects of the Invention

[0025] The research results of the present invention show that the recombinant type I humanized collagen can significantly inhibit the growth and proliferation ability (including DNA replication activity) of breast cancer cells (including both non-triple-negative breast cancer cells and triple-negative breast cancer cells), and can inhibit the migration characteristics of tumor cells (including both non-triple-negative breast cancer cells and triple-negative breast cancer cells). At the same time, it can also promote breast cancer cells to enter dormancy, thus laying a foundation for the preparation of drugs for treating breast cancer, and also providing a certain theoretical basis for the safety and effectiveness of recombinant type I humanized collagen in clinical use for breast cancer patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is the statistical result of the influence of recombinant type I humanized collagen at different concentrations on the proliferation activity of MCF-7 cells at different times.

[0027] Figure 2 It is the statistical result of the influence of recombinant type I humanized collagen at different concentrations on the proliferation activity of MDA-MB-231 cells at different times.

[0028] Figure 3 It is the statistical result of the influence of recombinant type I humanized collagen at different concentrations on the proliferation activity of SK-BR-3 cells at different times.

[0029] Figure 4Statistical results of the effect of the recombinant type I humanized collagen provided by the present invention on the migration ability of MCF-7 cells at different times.

[0030] Figure 5 Statistical results of the effect of the recombinant type I humanized collagen provided by the present invention on the migration ability of MDA-MB-231 cells at different times.

[0031] Figure 6 Statistical results of the effect of the recombinant type I humanized collagen provided by the present invention on the migration ability of SK-BR-3 cells at different times.

[0032] Figure 7 Detection results of the effect of the recombinant type I humanized collagen provided by the present invention on the DNA replication activity of MCF-7 cells in the EDU cell proliferation experiment.

[0033] Figure 8 Detection results of the effect of the recombinant type I humanized collagen provided by the present invention on the DNA replication activity of MDA-MB-231 cells in the EDU cell proliferation experiment.

[0034] Figure 9 Detection results of the effect of the recombinant type I humanized collagen provided by the present invention on the DNA replication activity of SK-BR-3 cells in the EDU cell proliferation experiment.

[0035] Figure 10 Detection results of the effect of the recombinant type I humanized collagen provided by the present invention on the cell cycle of MDA-MB-231 cells. Detailed implementation manners

[0036] The following describes the implementation manners of the present invention, but the present invention is not limited thereto.

[0037] In the present invention, the meaning expressed by "can" includes both the meaning of performing a certain process and not performing a certain process.

[0038] In the present invention, "optional" or "optionally" means that the subsequent described event or situation may or may not occur, and this description includes the situation where the event occurs and the situation where the event does not occur.

[0039] In the present invention, the terms "comprise", "have", "include" or "contain" can mean inclusive or open-ended, and do not exclude additional, unrecited elements or method steps. At the same time, "comprise", "have", "include" or "contain" can also mean closed-ended, excluding additional, unrecited elements or method steps.

[0040] In the present invention, the "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", etc. mentioned refer to the specific elements (e.g., features, structures, properties, and / or characteristics) related to the described embodiment, which are included in at least one of the embodiments described herein, and may or may not be present in other embodiments. Additionally, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0041] In the present invention, the numerical range represented by "numerical value A to numerical value B" or "numerical value A - numerical value B" refers to the range including the endpoint numerical values A and B.

[0042] In the present invention, the term "a", "an", or "the" may refer to "one", or may also refer to "one or more", "at least one", and "one or more than one".

[0043] In the present invention, the terms "individual", "patient", or "subject" include mammals. Mammals include, but are not limited to, domestic animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non - human primates such as monkeys), rabbits, and rodents (e.g., mice and rats).

[0044] In the present invention, "treatment" means that after a subject suffers from a disease, the subject is contacted (e.g., administered) with the recombinant type I humanized collagen described in the present invention, so that the symptoms of the disease are alleviated compared with the situation without contact, and it does not necessarily mean complete suppression of the symptoms of the disease. Suffering from a disease means that the body shows symptoms of the disease.

[0045] In the present invention, "prevention" means that before a subject suffers from a disease, by contacting the subject with the recombinant type I humanized collagen described in the present invention, the probability of suffering from the disease and / or the symptoms after suffering from the disease are reduced compared with the situation without contact, and it does not necessarily mean complete suppression of getting the disease.

[0046] In the present invention, the recombinant type I humanized collagen is described in the Chinese patent application with the application number 202110968550.2 and the invention title "A Recombinant Type I Humanized Collagen Polypeptide and Its Preparation Method and Use".

[0047] In some embodiments, the recombinant humanized type I collagen of the present invention comprises n repeats of the sequence shown in SEQ ID No.1 (composed of 60 amino acids, from the N-terminus to the C-terminus are GEKGS PGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQ GPSGAS), where n is an integer greater than or equal to 1. When n is an integer greater than or equal to 2, the repeat sequences are directly linked; optionally, the N-terminus of the recombinant humanized type I collagen comprises an amino acid sequence that can be excised by TEV protease.

[0048] In some preferred embodiments, the recombinant humanized type I collagen of the present invention comprises n repeats of the sequence shown in SEQ ID No.1, where n is an integer greater than or equal to 1. When n is an integer greater than or equal to 2, the repeat sequences are directly linked; and the N-terminus of the recombinant humanized type I collagen comprises an amino acid sequence that can be excised by TEV protease, and the amino acid sequence that can be excised by TEV protease comprises the sequence shown in SEQ ID No.2 (composed of 6 amino acids, from the N-terminus to the C-terminus are ENLYFQ).

[0049] In some more preferred embodiments, the recombinant humanized type I collagen of the present invention comprises n repeats of the sequence shown in SEQ ID No.1, where n is 4, the repeat sequences are directly linked, and the N-terminus of the recombinant humanized type I collagen comprises the sequence shown in SEQ ID No.2, and the sequence shown in SEQ ID No.1 and the sequence shown in SEQ ID No.2 are directly linked.

[0050] In some other preferred embodiments, the recombinant humanized type I collagen of the present invention comprises n repeats of the sequence shown in SEQ ID No.1, where n is an integer greater than or equal to 1. When n is an integer greater than or equal to 2, the repeat sequences are directly linked.

[0051] In some other more preferred embodiments, the recombinant humanized type I collagen of the present invention comprises n repeats of the sequence shown in SEQ ID No.1, where n is 4, the repeat sequences are directly linked, that is, it comprises the sequence shown in SEQ ID No.3.

[0052] GEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGASGEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGASGEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGASGEKGSPGADGPAGAPGTPGPQGIAGQRGVVGLPGQRGERGFPGLPGPSGEPGKQGPSGAS(SEQ ID No.3).

[0053] In the present invention, amino acid addition refers to adding amino acids to the C-terminus or N-terminus of an amino acid sequence, such as SEQ ID No.3, as long as the recombinant humanized type I collagen of the present invention retains the activity of the amino acid sequence of SEQ ID No.3.

[0054] In the present invention, amino acid substitution refers to replacing an amino acid residue at a certain position in an amino acid sequence, such as the sequence of SEQ ID No.3, with another amino acid residue, as long as the recombinant humanized type I collagen of the present invention retains the activity of the amino acid sequence of SEQ ID No.3.

[0055] In the present invention, amino acid insertion refers to inserting amino acid residues at appropriate positions in an amino acid sequence, such as the sequence of SEQ ID No.3. The inserted amino acid residues may also be adjacent to each other in whole or in part, or none of the inserted amino acids are adjacent to each other, as long as the recombinant humanized type I collagen of the present invention retains the activity of the amino acid sequence of SEQ ID No.3. In this article, the insertion position of amino acids is not between each repeat sequence.

[0056] In the present invention, amino acid deletion refers to deleting 1, 2 or more than 3 amino acids from an amino acid sequence, such as the sequence of SEQ ID No.3, as long as the recombinant humanized type I collagen of the present invention retains the activity of the amino acid sequence of SEQ ID No.3.

[0057] In the present invention, the amino acid substitution may be a conservative amino acid substitution, which means that compared with the amino acid sequence of SEQ ID No. 3, 3, more preferably 2 or 1 amino acid is replaced by an amino acid with similar or close properties to form a peptide. These conservative variant peptides can be generated according to the following amino acid substitutions: substitution of Ala by Val, Leu or Ile; substitution of Arg by Lys, Gln, Asn or His; substitution of Asn by Gln, His, Lys or Arg; substitution of Asp by Glu or Asn; substitution of Cys by Ser or Ala; substitution of Gln by Asn or Glu; substitution of Glu by Asp or Gln; substitution of Gly by Ala; substitution of His by Asn, Lys, Gln or Arg; substitution of Ile by Leu, Met, Ala, Val, Phe or norleucine; substitution of Leu by Ile, Met, Ala, Val, Phe or norleucine; substitution of Lys by Asn, Gln or Arg; substitution of Met by Ile, Leu or Phe; substitution of Phe by Leu, Val, Ile, Ala or Tyr; substitution of Pro by Ala; substitution of Ser by Thr; substitution of Thr by Ser or Val; substitution of Trp by Phe or Tyr; substitution of Tyr by Trp, Phe, Thr or Ser; and substitution of Val by Phe, Ala, Met, Ile, Leu or norleucine. The amino acid substitution may also be a non-conservative amino acid substitution.

[0058] As used herein, the terms "moderate stringency conditions", "medium-high stringency conditions", "high stringency conditions" or "very high stringency conditions" describe the conditions for nucleic acid hybridization and washing. Guidance for performing hybridization reactions can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6, which is incorporated herein by reference. Both aqueous and non-aqueous methods are described in this reference and either can be used. For example, specific hybridization conditions are as follows: (1) Low stringency hybridization conditions are in 6× sodium chloride / sodium citrate (SSC) at about 45°C, then at least 50°C, washing twice in 0.2× SSC, 0.1% SDS (for low stringency conditions, the wash temperature can be increased to 55°C); (2) Moderate stringency hybridization conditions are in 6× SSC at about 45°C, then at 60°C, washing once or more in 0.2× SSC, 0.1% SDS; (3) High stringency hybridization conditions are in 6× SSC at about 45°C, then at 65°C, washing once or more and preferably in 0.2× SSC, 0.1% SDS; (4) Very high stringency hybridization conditions are 0.5 M sodium phosphate, 7% SDS, at 65°C, then at 65°C, washing once or more in 0.2× SSC, 1% SDS.

[0059] Unless otherwise defined, all other technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0060] Examples

[0061] The present invention is further illustrated by the following examples, but any example or combination thereof should not be construed as limiting the scope or implementation of the present invention. The scope of the present invention is defined by the appended claims, and one of ordinary skill in the art can clearly understand the scope defined by the claims in combination with this specification and general common knowledge in the art. Without departing from the spirit and scope of the present invention, those skilled in the art can make any modifications or changes to the technical solutions of the present invention, and such modifications and changes are also included within the scope of the present invention.

[0062] For those not specifically noted in the examples, the operations were carried out under conventional conditions or conditions recommended by the manufacturer. All reagents or instruments not indicating the manufacturer were conventional products available commercially. To better illustrate the present invention, numerous specific details are given in the following detailed description. Those skilled in the art should understand that the present invention can be implemented without some of these specific details. In other examples, well-known methods, means, equipment and steps in the art are not described in detail so as to highlight the gist of the present invention.

[0063] Experimental materials:

[0064] Breast cancer cells: MCF-7 cells, MDA-MB-231 cells, and SK-BR-3 cells are from ATCC.

[0065] FBS is from: Sigma.

[0066] Serum-free medium is from: Sigma.

[0067] The amino acid sequence of recombinant humanized type I collagen is shown in SEQ ID No.3.

[0068] CCK-8 kit is from: GLPbio.

[0069] EDU and cell cycle detection kits are from: Beyotime.

[0070] Example 1: Effect of recombinant humanized type I collagen on cell proliferation

[0071] Experimental method:

[0072] Experimental group: Breast cancer cells + 10% FBS medium + recombinant humanized type I collagen co-culture.

[0073] Negative control group: Breast cancer cells + 10% FBS medium, co-cultured without collagen.

[0074] Blank control group: Without cells, 10% FBS medium, co-cultured without collagen.

[0075] After setting up the experimental groups, inoculate three types of breast cancer cells (MCF-7 cells, MDA-MB-231 cells, and SK-BR-3 cells) at an appropriate density according to the need into 96-well plates, with a volume of 100 μL / well. After the cells adhere, add recombinant humanized type I collagen with final concentrations of 0.01 mg / mL, 0.1 mg / mL, and 1 mg / mL (labeled as 0.01 mg / mL, 0.1 mg / mL, and 1 mg / mL in the figure), and the negative control group does not add collagen (labeled as 0 mg / mL in the figure); after co-culturing the cells with collagen for 24 h and 48 h, transfer the 96-well plates to the cell culture hood in the cell room, discard the medium, aspirate the residual liquid, add 100 μL of serum-free medium and 10 μL of CCK-8 to each well under light protection conditions, and add PBS around the 96-well plates to prevent reagent evaporation; place the 96-well plates in an incubator at 37 °C for light-protected culture for 1 hour, and use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance value OD 450nm, measure 3 times repeatedly and take the average value. Calculate according to the formula in the CCK-8 kit instruction manual: Cell survival rate = (OD value of experimental group - OD value of blank control group) / (OD value of negative control group - OD value of blank control group) × 100%.

[0076] Experimental results:

[0077] The results are as Figures 1 to 3 shown that recombinant humanized collagen type I can inhibit the proliferation activity of breast cancer cells.

[0078] After grouped culture, compared with the control group, the proliferation activities of the three kinds of breast cancer cells treated with recombinant humanized collagen type I were significantly reduced, indicating that recombinant humanized collagen type I has an inhibitory effect on the proliferation ability of breast cancer cells.

[0079] Example 2: Effect of recombinant humanized collagen type I on cell migration

[0080] Experimental method:

[0081] The experimental grouping was the same as in Example 1.

[0082] Inoculate breast cancer cells into six-well plates at a volume of 2 mL / well. After the cells adhered, add recombinant humanized collagen type I prepared with 10% FBS medium to the experimental group for co-culture (marked as the experimental group in the figure), with a final concentration of collagen of 1 mg / mL. The control group (marked as the control group in the figure) was added with 10% FBS medium. When the cells in each well reached 95% confluence to form a monolayer of cells, use a 200 μL pipette tip to make a "one"-shaped scratch on the monolayer of cells, wash 3 times with PBS, continue to add recombinant humanized collagen type I prepared with serum-free medium to the experimental group for culture, with a final concentration of collagen of 1 mg / mL, and the control group was added with serum-free medium. After incubating in the incubator for 24 h, aspirate the culture medium, wash 3 times with PBS, observe and take pictures under an inverted fluorescence microscope, and at the same time count the relative cell migration rate.

[0083] Experimental results:

[0084] The results are as Figures 4 to 6 shown that recombinant humanized collagen type I can inhibit the migration ability of breast cancer cells.

[0085] After grouped culture, compared with the control group, the migration rates of the three kinds of breast cancer cells treated with recombinant humanized collagen type I were significantly decreased, indicating that recombinant humanized collagen type I has an inhibitory effect on the migration ability of breast cancer cells.

[0086] Example 3: Experimental method for the effect of recombinant humanized collagen type I on cell DNA replication activity:

[0087] The experimental grouping was the same as in Example 1, and the final concentration of collagen co-culture was 1 mg / mL.

[0088] Breast cancer cells in the logarithmic growth phase were seeded into a 96-well plate at a density of 8×10 3 cells per well. The next day, after adherent culture to the normal growth stage, staining was performed using an EDU kit. First, an equal volume of pre-warmed 1× EdU working solution (10 μM) at 37 °C was added to the 6-well plate and incubated in an incubator at 37 °C for 2 h; after the EdU-labeled cells were completed, the culture medium was removed, and 0.1 mL of 4% (W / V) paraformaldehyde was added for fixation. After fixation at room temperature for 15 min, the fixing solution was removed, and the cells in each well were washed 3 times with 0.1 mL of washing solution, 3 - 5 minutes each time. After removing the washing solution, 0.1 mL of permeabilization solution (PBS containing 0.3% (V / V) Triton X-100) was added to each well and incubated at room temperature for 10 - 15 minutes. After removing the permeabilization solution, the cells in each well were washed 1 - 2 times with 0.1 mL of washing solution, 3 - 5 minutes each time, and then 0.05 mL of Click reaction solution was added to each well. After incubating at room temperature in the dark for 30 minutes, the Click reaction solution was aspirated, and the cells were washed 3 times with the washing solution, 3 - 5 minutes each time. The cell nuclei were stained with Hoechst 33342 and incubated at room temperature in the dark for 10 minutes. Fluorescence detection could then be performed.

[0089] Experimental results:

[0090] The results were as Figures 7 to 9 shown. Recombinant humanized type I collagen could inhibit the DNA replication activity of breast cancer cells and further inhibit the proliferation ability of breast cancer cells.

[0091] After grouped culture, compared with the control group, the proportion of breast cancer cells (red fluorescence) in the proliferative activity after treatment with recombinant humanized type I collagen decreased significantly, indicating that recombinant humanized type I collagen had an inhibitory effect on the DNA replication activity and proliferation of breast cancer cells.

[0092] Example 4: Effect of recombinant humanized type I collagen on the cell cycle

[0093] Experimental method:

[0094] The experimental grouping was the same as in Example 1, and the final concentration of collagen co-culture was 1 mg / mL.

[0095] After digesting MDA-MB-231 breast cancer cells with trypsin, add them to 1 mL of 70% ethanol pre-cooled in an ice bath, gently pipette and mix well, and fix at 4°C for 30 minutes. Centrifuge at 1000 g for 3 - 5 minutes to precipitate the cells. Add 1 mL of PBS pre-cooled in an ice bath to resuspend the cells. Centrifuge again to precipitate the cells, aspirate the supernatant, add 0.5 mL of propidium iodide staining solution to each tube of cell sample, slowly and thoroughly resuspend the cell pellet, and incubate at 37°C in the dark for 30 minutes to complete flow cytometry detection.

[0096] Experimental results:

[0097] The results are as Figure 10 shown. Recombinant type I humanized collagen can affect the cell division cycle of breast cancer cells.

[0098] After grouped culture, compared with the control group, after treatment with recombinant type I humanized collagen, the proportion of breast cancer cells in the dormant cycle (G1 / G0 phase) increased significantly, and the proportion of breast cancer cells in the proliferative active cycle (G2 / M phase) decreased significantly, indicating that recombinant type I humanized collagen can promote breast cancer cells to enter the dormant cycle (G1 / G0 phase).

[0099] Summary:

[0100] Breast cancer currently ranks first in the global cancer incidence, and the treatment methods are limited. Especially for triple-negative breast cancer, the treatment means are single, and the recurrence and metastasis rates are high. There is an urgent need for more effective and safe treatment methods. Recombinant type I humanized collagen has the characteristics of high water solubility and high biological activity. Existing experiments have confirmed that recombinant type I humanized collagen can inhibit the growth and proliferation ability of breast cancer cells, and can also inhibit the migration characteristics of tumor cells, and its mechanism may depend on promoting tumor cells to enter the dormant cycle. This will provide a certain theoretical basis for the safety and effectiveness of recombinant type I humanized collagen in clinical application for breast cancer patients.

Claims

1. Use of recombinant humanized type I collagen in the preparation of a medicament for treating breast diseases; Among them, The amino acid sequence of the recombinant humanized type I collagen is as shown in SEQ ID No. 3; The breast disease is breast cancer, and the breast cancer is luminal A breast cancer, HER-2 overexpressing breast cancer, basal-like breast cancer or triple-negative breast cancer.

Images