CN112980951A - Application of mitochondrial protein SLC25A24 in diagnosis and prognosis judgment of colorectal cancer - Google Patents
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Abstract
The invention discloses application of a mitochondrial protein SLC25A24 in diagnosis and prognosis judgment of colorectal cancer. In a first aspect of the application, there is provided the use of a reagent for the quantitative determination of SLC25a24 in the manufacture of a product for the diagnostic and/or prognostic determination of colorectal cancer. According to the application of the embodiment of the application, at least the following beneficial effects are achieved: the applicants have surprisingly found that the mitochondrial protein and its mRNA are significantly differentially expressed between patients with colorectal cancer and normal persons, and therefore the SLC25A24 can be detected at the nucleic acid and/or protein level in subjects to effectively diagnose or judge whether colorectal cancer is caused and the prognosis of colorectal cancer.
Description
Technical Field
The application relates to the field of cancer detection, in particular to application of a mitochondrial protein SLC25A24 in diagnosis and prognosis of colorectal cancer.
Background
Colorectal cancer is one of the most fatal cancers in the world. Despite the recent emergence of new therapeutic approaches, such as targeted therapy and immunotherapy, the mortality rate for colorectal cancer remains high and about 50% of patients experience relapse and distant metastasis during treatment. The incidence and mortality of colorectal cancer in China are obviously higher than the average level in the world, and the incidence and mortality of colorectal cancer in China are increased by 84.1% and 8.2% from 1990 to 2017. Therefore, based on the current diagnosis and treatment situation of Chinese colon cancer, it is necessary to develop the basic research work related to colorectal cancer. Screening for early diagnostic markers and therapeutic targets associated with colorectal cancer is an important part of basic research efforts.
Mitochondria, as an important organelle in eukaryotes, are highly involved in the occurrence and development of colorectal cancer. In the early stages, metabolic changes caused by mitochondrial depletion are the basis of cellular transformation. As the tumor progresses, mitochondrial function is reactivated to accommodate cell proliferation and metastasis. Mitochondria play an indispensable role in energy production, apoptosis and metabolism, and the signal pathway regulation mechanism related to the mitochondria is gradually a research hotspot in the occurrence and development of colorectal cancer. However, the work of studying targets and biomarkers for diagnosis and prognosis of colorectal cancer using mitochondria as a subject has been less advanced, and there is a lack of markers that can effectively evaluate colorectal cancer.
Disclosure of Invention
The present application is directed to solving at least one of the problems in the prior art. Therefore, the application provides the application of the mitochondrial protein SLC25A24 capable of effectively evaluating the colorectal cancer in the diagnosis and prognosis judgment of the colorectal cancer.
In a first aspect of the application, there is provided the use of a reagent for the quantitative determination of SLC25a24 in the manufacture of a product for the diagnostic and/or prognostic determination of colorectal cancer.
According to the application of the embodiment of the application, at least the following beneficial effects are achieved:
the applicants have surprisingly found that the mitochondrial protein and its mRNA are significantly differentially expressed between patients with colorectal cancer and normal persons, and therefore the SLC25A24 can be detected at the nucleic acid and/or protein level in subjects to effectively diagnose or judge whether colorectal cancer is caused and the prognosis of colorectal cancer.
Among them, the mitochondrial protein SLC25a24(Calcium-binding mitogenic carrier protein SCaMC-1) is a Calcium-binding mitochondrial carrier protein SCaMC-1, encoded by SLC25a24 gene, and mainly participates in mediating the reversible electroneutral exchange of Mg-ATP or Mg-ADP with phosphate ions as a Calcium-dependent mitochondrial solute carrier, catalyzing the net uptake or efflux of adenine nucleotides on the inner mitochondrial membrane.
Diagnosis refers to the judgment of whether a subject suffers from colorectal cancer, and prognosis refers to the judgment of the possible course and outcome of colorectal cancer, and specifically may include the judgment of the specific outcome (such as recovery, appearance or disappearance of other abnormalities such as specific symptoms, signs and complications, and death) of colorectal cancer.
In some embodiments of the present application, the colorectal cancer is selected from at least one of a colon adenoma, a rectal adenoma, a colon adenocarcinoma, a caecum adenocarcinoma, a rectal adenocarcinoma, a sigmoid colon adenocarcinoma, a mucinous colon adenocarcinoma, and a mucinous rectal adenocarcinoma.
In some embodiments of the present application, the agent quantitatively detects SLC25a24 at the nucleic acid and/or protein level. Since the nucleic acid sequence as well as the amino acid sequence of SLC25a24 are known in the art, one skilled in the art can prepare reagents capable of quantitatively detecting SLC25a24 based on conventional means or by commercially available reagents.
In some embodiments of the present application, the reagent is selected from the group consisting of a PCR reagent, an immunohistochemical reagent, a western blot reagent. The PCR reagent specifically includes a primer capable of specifically amplifying a nucleic acid molecule fragment expressing a gene encoding the mitochondrial protein, and may further include an enzyme mixture for carrying out an amplification reaction, dNTP, a buffer solution, and the like; detection probes or other components may also be included, depending on the particular PCR method. Specifically, the immunohistochemical reagent includes an antibody capable of specifically binding to the mitochondrial protein, and for detection, a detectable label such as a radioisotope, a fluorescent group, an enzyme, a dye, or the like may be further coupled to the antibody.
In some embodiments of the present application, the level of SLC25a24 is negatively correlated with an indicator of survival of the subject. That is, the lower the content/expression of SLC25a24 nucleic acid or protein, the worse the prognosis of the subject. For example, the level of SLC25a24 in the subject is compared to a predetermined control level and if the comparison indicates that the level of SLC25a24 in the subject is below the control level, the subject is indicated to have colorectal cancer, or a poor prognosis.
In some embodiments of the present application, the survival index is selected from at least one of survival, disease progression.
In a second aspect of the application, a kit is provided, comprising reagents for the quantitative detection of SLC25a 24.
In some embodiments of the present application, the kit is for use in the diagnosis and/or prognosis of colorectal cancer.
In some embodiments of the present application, the colorectal cancer is selected from at least one of a colon adenoma, a rectal adenoma, a colon adenocarcinoma, a caecum adenocarcinoma, a rectal adenocarcinoma, a sigmoid colon adenocarcinoma, a mucinous colon adenocarcinoma, and a mucinous rectal adenocarcinoma.
In some embodiments of the present application, the agent quantitatively detects SLC25a24 at the nucleic acid and/or protein level.
Additional aspects and advantages of the present application will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the present application.
Drawings
FIG. 1 shows the results of screening for prognostic factors in example 1 of the present application.
Figure 2 is the differential expression validation result of SLC25a24 of example 2 of the present application.
Figure 3 is a quantitative comparison of the expression of SLC25a24 of example 2 of the present application.
FIG. 4 is a comparison of the staging according to different degrees of development of colorectal cancer in example 3 of the present application versus the change in SLC25A24 expression in stage I-IV patients.
Figure 5 is a comparison of the overall survival of different expression levels of SLC25a24 in example 3 of the present application.
FIG. 6 is a comparison of progression-free survival for different expression levels of SLC25A24 in example 3 of the present application.
FIG. 7 is a comparison of SLC25A24 expression levels in colorectal cancer patients of different subtypes in example 3 of the present application.
Detailed Description
The conception and the resulting technical effects of the present application will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, and not all embodiments, and other embodiments obtained by those skilled in the art without inventive efforts based on the embodiments of the present application belong to the protection scope of the present application.
The following detailed description of embodiments of the present application is provided for the purpose of illustration only and is not intended to be construed as a limitation of the application.
In the description of the present application, the meaning of a plurality is one or more, the meaning of a plurality is two or more, and the above, below, exceeding, etc. are understood as excluding the present number, and the above, below, within, etc. are understood as including the present number. If the first and second are described for the purpose of distinguishing technical features, they are not to be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of the technical features indicated.
In the description of the present application, reference to the description of the terms "one embodiment," "some embodiments," "an illustrative embodiment," "an example," "a specific example," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
Example 1
Screening for differentially expressed mitochondrial proteins
Cancer tissues of 8 colorectal cancer patients (the diagnosis was confirmed by histopathological examination) and paracancerous normal tissues as controls were collected.
Extracting proteins from the tissue sample. Every two patients in the same disease period are mixed into one sample, namely 4 tumor group samples and 4 paracancer group samples are subjected to mass spectrometry, and proteins with significant differences in expression amount are screened out. And enriching the differential expression protein by GO, KEGG, subcellular structure and other enrichment strategies to obtain the mitochondria-related protein with significant changes in expression. The results show that the expression of the mitochondrial related protein in colorectal cancer patients is significantly changed compared with that of normal people, and the subcellular structural analysis shows that the mitochondrial protein is most obviously reduced in the protein with the reduced expression. While 164 proteins were reduced in 192 mitochondrial proteins with altered expression.
538 transcriptome data and clinical data of colorectal Cancer patients are downloaded from The Cancer Genome Atlas Program (TCGA) database, and 11 independent prognostic factors including SLC25A24 and other mitochondrial genes are screened from The results by combining proteomic results and a bioinformatics method. The results are shown in FIG. 1. N1-N4 represent paracarcinoma groups, and C1-C4 represent tumor groups. As can be seen in figure 1, there is a clear down-regulation of the SLC25a24 mitochondrial gene in the tumor group.
Example 2
Differential expression verification experiment
Clinical information of colorectal Cancer patients in The Cancer Genome Atlas Program (TCGA) database is searched, patients with other malignant tumor history and receiving radiotherapy or chemotherapy are excluded, 538 relevant data of colorectal Cancer patients are screened out to be used as an experimental group for research, and 51 relevant data of normal people are selected to be used as a control group. The expression of SLC25A24 in the experimental group and the control group were statistically analyzed. Immunohistochemical results of SLC25A24 Protein distribution expression in tumor tissues and normal Human tissues of 21 colorectal cancer patients in The Human Protein Atlas database were retrieved for analysis.
As shown in FIG. 2, A is the expression statistics of the corresponding mitochondrial proteins, and B and C are the immunohistochemical results of the control and experimental groups of mitochondrial proteins. In A of FIG. 2, the control group is high-expressing, and the experimental group is high-expressing and high-expressing, respectively, from top to bottom. From the immunohistochemical staining results of B and C, it can be seen that the control group had extensive strong positive staining, whereas the experimental group had weak staining. The above results indicate that mitochondrial proteins have significant down-regulated expression in the experimental group.
Qualitative results of the expression of mRNA of the mitochondrial protein SLC25a24 are shown in fig. 3, and it can be seen that there was significant down-regulated expression of mRNA of this mitochondrial protein in both experimental groups compared to the control group (p < 0.01).
The above results indicate that SLC25A24 has significant differential expression in colorectal cancer patients, and has potential as a diagnostic or prognostic marker.
Example 3
The value of the prognosis for SLC25A24 in colorectal cancer was further validated as follows:
using the relevant clinical data of 538 patients with colorectal cancer screened in example 2, the above-mentioned changes in the mRNA expression of mitochondrial proteins in patients in stages I to IV were analyzed according to their stages of different degrees of development, and the results are shown in FIG. 4. It can be seen from the figure that the expression of SLC25a24 gradually decreased with the progression of colorectal cancer, and there was a significant difference in SLC25a24 expression between stage i and stage iv patients (p < 0.01).
The overall survival profile and progression-free survival profile were plotted from the relevant clinical data for 538 colorectal cancer patients, and the results are shown in fig. 5 and 6, respectively, in which the upper side is a high-expression curve and the lower side is a low-expression curve. As can be seen in fig. 5 and 6, the expression level of SLC25a24 was negatively correlated with the overall survival of the patients (p <0.01), while also being negatively correlated with disease progression (p < 0.01).
Searching clinical information of colorectal cancer patients in an oncomine database, excluding patients who have other malignant tumor histories and received radiotherapy or chemotherapy, screening out relevant data of 410 colorectal cancer patients, and respectively counting the expression conditions of SLC25A24 of various subtypes of colorectal cancer, wherein the results are shown in figure 7. In the above, nos. 1 to 8 represent colon adenomas (30 cases), rectal adenomas (7 cases), colon adenocarcinomas (142 cases), cecal adenocarcinoma (39 cases), rectal adenocarcinoma (134 cases), sigmoid colon adenocarcinoma (13 cases), mucinous colon adenocarcinoma (35 cases), and mucinous colon adenocarcinoma (10 cases), respectively. As can be seen, SLC25a24 showed significant down-regulation in a number of colorectal tumor subtypes.
The results further indicate that the SLC25A24 has higher value as a prognosis marker of colorectal cancer.
The present application has been described in detail with reference to the embodiments, but the present application is not limited to the embodiments described above, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present application. Furthermore, the embodiments and features of the embodiments of the present application may be combined with each other without conflict.
Claims (10)
1. Application of a reagent for quantitatively detecting SLC25A24 in preparing a product for diagnosing and/or prognostically judging colorectal cancer.
2. The use according to claim 1, wherein the colorectal cancer is selected from at least one of a colon adenoma, a rectum adenoma, a colon adenocarcinoma, a cecal adenocarcinoma, a rectum adenocarcinoma, a sigmoid colon adenocarcinoma, a mucinous colon adenocarcinoma, and a mucinous rectum adenocarcinoma.
3. The use of claim 1, wherein the agent quantitatively detects SLC25A24 at the nucleic acid and/or protein level.
4. Use according to claim 3, wherein the reagents are selected from PCR reagents, immunohistochemical reagents, Western blotting reagents.
5. The use of claim 1, wherein the level of SLC25a24 is negatively correlated with an indicator of survival of the subject.
6. The use of claim 5, wherein the survival index is selected from at least one of survival, disease progression.
7. The kit is characterized by comprising a reagent for quantitatively detecting SLC25A 24.
8. The kit according to claim 7, wherein the kit is for use in the diagnosis and/or prognosis of colorectal cancer.
9. The kit of claim 8, wherein the colorectal cancer is selected from at least one of colon adenoma, rectal adenoma, colon adenocarcinoma, cecal adenocarcinoma, rectal adenocarcinoma, sigmoid colon adenocarcinoma, mucinous colon adenocarcinoma, and mucinous rectal adenocarcinoma.
10. The kit of claim 7, wherein the reagents allow for the quantitative determination of SLC25A24 at the nucleic acid and/or protein level.
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