Application of MUC-24 in preparation of drugs for regulating and controlling proliferation cycle and apoptosis of breast cancer cells

Abstract

The invention is applicable to the technical field of biological medicines, and provides application of MUC-24 in preparation of medicines for regulating and controlling proliferation cycle and apoptosis of breast cancer cells. The invention firstly determines that the MUC-24 is remarkably high in expression in breast cancer tissues and is closely related to prognosis of patients, and prompts that the MUC-24 can be used as a potential diagnosis, treatment and prognosis monitoring target of the breast cancer. By targeting MUC-24, the apoptosis of breast cancer cells can be activated, and the proliferation capacity and cell viability of the breast cancer cells can be remarkably inhibited, so that a good anti-tumor effect is shown. Further research finds that MUC-24 induces breast cancer cell cycle arrest and apoptosis by regulating and controlling extracellular matrix, and enriches cognition on the mechanism of regulating and controlling breast cancer cell proliferation molecules by MUC-24. Theoretical support is provided for clinical application of MUC-24 as a new molecular treatment target, and a new thought and a new direction are provided for finding a new target for breast cancer treatment.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the application of MUC-24 in the preparation of drugs for regulating the proliferation cycle and apoptosis of breast cancer cells. Background Art

[0002] The latest global cancer data released by the World Health Organization's International Agency for Research on Cancer show that breast cancer has become the most common malignant tumor in women and the most common type of cancer worldwide. Molecular markers play a key role in the diagnosis and treatment of breast cancer. According to the expression of molecular markers of estrogen or progesterone receptors and human epidermal growth factor 2 (ERBB2), breast cancer is divided into three main subtypes: hormone receptor positive / ERBB2 negative (70%), ERBB2 positive (15-20%) and triple negative (15%). At present, the treatment of breast cancer includes traditional treatment (surgery combined with radiotherapy and chemotherapy) and molecular targeted therapy (endocrine therapy, anti-HER-2 therapy). However, traditional treatment methods have poor prognosis for some patients due to their multidrug resistance and side effects. Molecular targeted therapy targets the marker molecules of tumor cells, and achieves the purpose of precise treatment of tumors by targeting and inhibiting the proliferation of tumor cells, interfering with the cell cycle, inducing tumor cell apoptosis and inhibiting tumor angiogenesis. Currently, commonly used targeted drugs include HER2 targeted drugs (such as trastuzumab) and hormone receptor targeted drugs (such as tamoxifen, aromatase inhibitors). With the advancement of genetic testing technology, new small molecule inhibitors targeting common breast cancer mutations, such as PI3K inhibitors, CDK4 / 6 inhibitors, and immune checkpoint inhibitors, are being developed. Targeted therapy based on the patient's molecular characteristics will significantly improve the effect of individualized treatment. In addition, molecular targeted therapy can be used in combination with other treatments (such as immunotherapy, chemotherapy, etc.) to enhance the overall treatment effect. Therefore, it is crucial to deeply explore the pathogenesis of breast cancer and find new molecular diagnostic and therapeutic targets.

[0003] MUC-24 is a transmembrane sialomucin and cell adhesion molecule that regulates the proliferation, adhesion, and migration of hematopoietic progenitor cells. Existing studies have mainly focused on the potential of MUC-24 as a molecular marker for metastasis in prostate cancer and colorectal cancer, and it has a certain regulatory effect in gliomas. However, no studies have reported its anti-tumor effect in breast cancer cells and its related mechanism. Therefore, the application of MUC-24 in the preparation of drugs that regulate the proliferation cycle and apoptosis of breast cancer cells is proposed. Summary of the invention

[0004] The purpose of the present invention is to provide the use of MUC-24 in the preparation of drugs for regulating the proliferation cycle and apoptosis of breast cancer cells, aiming to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] Application of MUC-24 in the preparation of drugs for regulating the proliferation cycle and apoptosis of breast cancer cells.

[0007] Furthermore, the MUC-24 regulates the cell cycle and apoptosis of breast cancer cells by regulating the extracellular matrix (ECM).

[0008] Compared with the prior art, the present invention has the following beneficial effects:

[0009] The present invention clarifies for the first time that MUC-24 is significantly highly expressed in breast cancer tissue and is closely related to the patient's prognosis, suggesting that MUC-24 can be used as a potential target for diagnosis, treatment and prognosis monitoring of breast cancer. By targeting MUC-24, breast cancer cell apoptosis can be activated, and the proliferation ability and cell viability of breast cancer cells can be significantly inhibited, thereby showing a good anti-tumor effect. Further studies have found that MUC-24 induces breast cancer cell cycle arrest and apoptosis by regulating extracellular matrix (ECM) remodeling, enriching the understanding of the molecular mechanism of MUC-24 regulating breast cancer cell proliferation. The present invention provides theoretical support for the clinical application of MUC-24 as a new molecular therapeutic target, and provides new ideas and directions for finding new targets for breast cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 The molecular mechanism diagram of the present invention.

[0011] Figure 2 The expression of MUC-24 in breast cancer tissue and its relationship with grade and prognosis; A is the expression of MUC-24 in breast cancer tissue and normal breast tissue detected by TCGA database; B is the relationship between MUC-24 expression and disease-free survival of breast cancer patients; C is the relationship between MUC-24 expression and overall survival of breast cancer patients.

[0012] Figure 3 To inhibit the effect of MUC-24 expression on breast cancer cell viability and proliferation; A is the Western Blot method to detect the expression level of MUC-24 in cells; B is the CCK-8 method to detect cell viability; C is the clone formation experiment to detect cell proliferation ability.

[0013] Figure 4 Cell cycle analysis was performed by flow cytometry.

[0014] Figure 5 Cell apoptosis was detected by flow cytometry.

[0015] Figure 6 To enrich and analyze the effect of MUC-24 on downstream signaling pathways.

[0016] Figure 7 To knock down MUC-24 to inhibit the proliferation of breast cancer cells in vivo; wherein, A is a schematic diagram of the tumor volume of each group; B is the HE toxicity detection of the internal organs (heart, liver, spleen, lung, and kidney) of each group of mice. DETAILED DESCRIPTION

[0017] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0018] The molecular mechanism diagram of the present invention is as follows Figure 1 As shown, MUC-24 regulates breast cancer cell cycle and apoptosis by regulating the extracellular matrix.

[0019] The specific implementation of the present invention is described in detail below in conjunction with specific embodiments.

[0020] Example 1: MUC-24 is highly expressed in breast cancer tissues and is associated with grade and prognosis;

[0021] First, the TCGA database was used for analysis, and the results showed that compared with normal breast tissue, the expression of MUC-24 in breast cancer tissue was significantly upregulated ( Figure 2 A); and is significantly negatively correlated with the prognosis of breast cancer patients ( Figure 2 In Figures B and C), patients with high MUC-24 expression (high) had a longer disease-free survival (DFS) than patients with low MUC-24 expression (low). Figure 2 B) and overall survival ( Figure 2 The above results suggest that MUC-24 may be a potential target for diagnosis, treatment and prognosis monitoring of breast cancer.

[0022] Example 2: Inhibition of MUC-24 can inhibit breast cancer cell proliferation and cell viability;

[0023] First, two effective siRNAs were designed to knock down the expression of MUC-24 in breast cancer cells, denoted as siMUC-24-1 and siMUC-24-2, respectively. Figure 3 As shown in A, compared with the control group (siNC), the MUC-24 knockdown groups (siMUC-24-1 and siMUC-24-2) significantly inhibited the expression of MUC-24, indicating that the knockdown efficiency was good. CCK-8 results showed ( Figure 3 (B), compared with the control group, the cell viability of the MUC-24 knockdown group was significantly inhibited. The results of the clone formation experiment showed that ( Figure 3 Middle C), compared with the control group, the number of cells in the MUC-24 knockdown group was significantly reduced. The above results suggest that inhibition of MUC-24 can significantly inhibit the proliferation ability and cell viability of breast cancer cells.

[0024] Example 3, inhibition of MUC-24 can induce breast cancer cell cycle arrest and apoptosis by regulating the extracellular matrix;

[0025] In order to explore the regulatory mechanism of MUC-24 on breast cancer cell proliferation, we used flow cytometry to detect the changes in breast cancer cell cycle and apoptosis before and after knocking down MUC-24. Figure 4 As shown in Figure 3, knockdown of MUC-24 expression induced cell cycle arrest in the S phase of breast cancer cells compared with the control group (siNC). Figure 5 As shown in the figure, compared with the control group, the number of apoptotic cells in the MUC-24 knockdown group (siMUC-24-1 and siMUC-24-2) increased significantly. The above results prove that inhibition of MUC-24 can induce cell cycle arrest and apoptosis in breast cancer cells and has a good anti-tumor effect. In order to find the mechanism of action of MUC-24, we enriched the downstream signaling pathways affected by MUC-24 by transcriptomics sequencing. The results showed that MUC-24 is mainly involved in regulating the remodeling process of extracellular matrix ( Figure 6 ).

[0026] Example 4: Regulatory effect of MUC-24 on breast cancer cells in vivo;

[0027] In order to detect the regulatory effect of MUC-24 on breast cancer cell proliferation in vivo, a breast cancer cell line 4T1 was used to establish a nude mouse breast cancer subcutaneous transplant tumor model, and the drug was treated for 21 days to observe the therapeutic effect of knocking down MUC-24 on breast cancer in nude mice. Figure 7 As shown in Figure A, compared with the control group (siNC), the MUC-24 knockdown group (siMUC-24) significantly inhibited the tumor volume, suggesting that MUC-24 knockdown can inhibit the proliferation of breast cancer cells in vivo. In addition, there was no obvious toxicity in the internal organs (heart, liver, spleen, lung, and kidney) of mice in each group ( Figure 7 Middle B) proves that siMUC-24 treatment has no obvious toxic side effects on nude mice. In summary, MUC-24 can regulate the proliferation of breast cancer cells in vivo.

[0028] Conclusion: This invention clearly shows for the first time that MUC-24 is significantly highly expressed in breast cancer tissues and is closely related to the prognosis of patients, suggesting that MUC-24 can be used as a potential target for diagnosis, treatment and prognosis monitoring of breast cancer. By targeting MUC-24, breast cancer cell apoptosis can be activated, and the proliferation ability and cell viability of breast cancer cells can be significantly inhibited, thereby showing a good anti-tumor effect. Further studies have found that MUC-24 induces breast cancer cell cycle arrest and apoptosis by regulating the extracellular matrix, enriching the understanding of the molecular mechanism of MUC-24 regulating breast cancer cell proliferation. This invention provides theoretical support for the clinical application of MUC-24 as a new molecular therapeutic target, and provides new ideas and directions for finding new targets for breast cancer treatment.

[0029] The above are only preferred embodiments of the present invention. It should be pointed out that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present invention. These should also be regarded as the protection scope of the present invention. These will not affect the effect of the implementation of the present invention and the practicality of the patent.

Claims

1. Application of MUC-24 in the preparation of drugs for regulating the proliferation cycle and apoptosis of breast cancer cells.

2. The use according to claim 1, characterized in that: The MUC-24 regulates the cell cycle and apoptosis of breast cancer cells by regulating the extracellular matrix.

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