CN113834927A - Salivary metabolism marker for predicting canceration of oral mucosa precancerous lesion patient and application thereof - Google Patents
Salivary metabolism marker for predicting canceration of oral mucosa precancerous lesion patient and application thereof Download PDFInfo
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Abstract
The invention discloses salivary metabolism markers for predicting canceration of patients with oral mucosa precancerous lesion and application thereof, wherein the group of salivary metabolism markers has important clinical significance for early diagnosis of oral squamous cell carcinoma, particularly prediction of the canceration of the oral mucosa precancerous lesion into cancer evolution. Specifically, the invention discloses metabolic markers which can be detected by CPSI-MS (compact peripheral component analysis-mass spectrometry) in situ mass spectrometry, wherein the metabolic markers comprise 4 metabolites Putrescine, Cadaverine, Thymidine and Adenosine which are gradually increased from healthy people to precancerous lesion to cancers, and 4 metabolites Hippuric acid, Phosphocholine, Glucose and Adrenic acid which are gradually decreased from healthy people to precancerous lesion to cancers, and when the 8 metabolites are up-regulated or down-regulated, a subject is a high-risk patient with cancerization tendency, and needs to pay close attention.
Description
Technical Field
The invention belongs to the field of biological medicines, and relates to a salivary metabolism marker and a diagnostic application thereof in the process of changing oral mucosa precancerous lesion into oral squamous cell carcinoma.
Background
Oral Squamous Cell Carcinoma (OSCC) is one of the most common types of head and neck malignancies. Worldwide, the incidence of oral squamous cell carcinoma is relatively higher in developing countries than in developed countries, and the incidence rate of oral squamous cell carcinoma is reported to be about 10 cases/10 ten thousand in the united states and more than 20 cases/10 ten thousand in india. In recent decades, the incidence of OSCC has been increasing year by year, and for example, in the above sea area, the incidence of female oral squamous cell carcinoma is 3.3/10 ten thousand, and the incidence of male squamous cell carcinoma is 3.8/10 ten thousand. Although the incidence rate of oral squamous cell carcinoma is not high compared with other tumors, the population of China is large, the patient base number is large, and the prevention and treatment of oral squamous cell carcinoma are both serious and far-going.
In recent decades, although the treatment technology and treatment concept of oral squamous cell carcinoma are continuously improved, the overall treatment effect is not improved significantly. The 5-year survival rate of oral squamous carcinoma patients is not significantly improved, still lingers around 60%, and even less than 30% of patients in advanced stages, the main reason being that most patients are already in advanced tumor stages at the time of diagnosis. The literature reports that the postoperative local recurrence rate of oral squamous cell carcinoma is 6.9-22%, and the survival rate of patients is obviously reduced once postoperative recurrence occurs. Previous studies have shown that the 3-year Overall Survival (OS) of patients with postoperative recurrence is about 52.6%, and the 5-year Overall Survival is only 24-50%.
Realizes early prevention, early diagnosis and early treatment of malignant tumors, and has positive significance for improving the treatment effect of malignant tumors. Oral squamous carcinoma is a malignant tumor derived from the epithelium of the oral mucosa, the pathological evolution of which includes normal mucosal epithelium-abnormally hyperplastic epithelium-malignant epithelial cells. Precancerous lesions of the oral mucosa mainly include oral leukoplakia and erythema, and the main pathological changes are abnormal hyperplasia of epithelial tissues (light, medium and severe abnormalities). The precancerous lesions (oral leukoplakia and erythema) of the oral mucosa are often seen in middle-aged and elderly people, patients have long course of disease and are not cured, and the greatest risk is that some patients can progress to oral squamous cell carcinoma, so that the patients are greatly psychologically burdened. Therefore, the method finds a marker capable of effectively predicting the malignant transformation risk of the oral mucosa precancerous lesion (oral leukoplakia and erythema), and has positive significance for improving the prevention and treatment of the oral mucosa precancerous lesion and the oral squamous cell carcinoma.
Disclosure of Invention
In order to realize early, quick and noninvasive screening of the canceration potential of the oral mucosa precancerous lesion, the invention aims to provide a group of salivary metabolism markers capable of accurately distinguishing healthy volunteers, oral mucosa precancerous lesion and oral squamous carcinoma patients.
The invention also aims to provide application of the salivary metabolism marker.
The technical scheme of the invention is as follows:
salivary metabolism markers for predicting canceration of patients with oral mucosa precancerous lesion comprise Putrescine Putrescine, Cadaverine Cadaverine, Thymidine Thymidine, Adenosine Adenosine, Hippuric acid, Phosphocholine Phosphocholine, Glucose or Adrenic acid.
Further, the marker: putrescine, Cadaverine, Thymidine and Adenosine are set as a group; the Hippuric acid, Phosphocholine, Glucose and Adrenic acid were set as another group, and the two groups were used in combination.
Furthermore, the metabolic marker is not only a cancerous metabolic marker of oral squamous cell carcinoma, but also can screen patients with oral mucosa precancerous lesion with high risk of canceration.
A kit for predicting the occurrence of canceration in a patient having precancerous lesions of the oral mucosa, said kit comprising a salivary metabolism marker as defined in any one of the preceding.
The application of the salivary metabolism marker in the preparation of a kit for predicting canceration of a patient with precancerous lesion of oral mucosa.
Further, the application: extracting a saliva sample for diagnosis, wherein the content of Putrescine, Cadaverine, Thymidine and Adenosine is gradually increased from a healthy person to a precancerous lesion patient to a cancer patient; the contents of Hippuric acid, Phosphocholine, Glucose and Adrenic acid are gradually decreased from healthy people-precancerous lesion patients-cancer patients.
Further, the metabolic marker not only diagnoses the oral squamous carcinoma patients, but also screens the oral mucosa precancerous lesion patients with high risk of canceration.
The invention has the following advantages:
(1) according to the invention, Putrescine (Putrescine), Cadaverine (Cadaverine), Thymidine (Thymidine), Adenosine (Adenosine), Hippuric acid (Hippuric acid), Phosphocholine (phosphorylcholine), Glucose (Glucose) and Adrenic acid (arachidonic acid) are selected as saliva metabolite molecular marker combinations, so that the change of the precancerous lesion of the oral mucosa to the cancer can be effectively predicted, and the precancerous lesion patients (white spots and erythema) of the oral mucosa with high risk of canceration are screened.
(2) The oral mucosa precancerous lesion patient (white spot and erythema) saliva diagnosis kit is developed based on Putrescine, cadoverine, thymine, Adenosine, Hippuric acid, Phosphocholine, Glucose and Adrenic acid, can realize early rapid and noninvasive screening of oral mucosa precancerous lesion patients (white spot and erythema) before malignant transformation, can meet the detection requirements of most medical institutions, and has a wide clinical application prospect.
In conclusion, the saliva marker can be used for screening oral mucosa precancerous lesion patients with high risk of canceration, is simple in method, non-invasive to operate and high in accuracy, and provides a new method and a new thought for early screening, early diagnosis and early treatment of oral squamous carcinoma and oral mucosa precancerous lesions.
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FIG. 1 shows the PLS-DA analysis results of healthy volunteers (HC), oral mucosal precancerous lesions (PML) and Oral Squamous Cell Carcinoma (OSCC) cases in the experimental development group (first group).
Figure 2 is a wien plot analysis of the common metabolite of the experimental development group (first batch) and validation group (second batch): 42 metabolites common to healthy volunteers (HC) vs oral mucosal precancerous lesions (PML) in the experimental development and validation groups; (b) 94 metabolites common to oral mucosal precancerous lesions (PML) vs Oral Squamous Cell Carcinoma (OSCC) in both experimental development and validation groups.
Fig. 3 is a schematic diagram of ROC curve analysis of all 373 cases with 8 markers as a combination, which can accurately distinguish early healthy volunteers vs precancerous patients (AUC 0.89), precancerous patients vs oral squamous carcinoma patients (AUC 0.83), and healthy volunteers vs oral squamous carcinoma patients (AUC 0.96).
Detailed Description
The invention is further described below with reference to the figures and examples.
1. The first embodiment is as follows: clinical case data for the study were included.
In this example a total of 373 saliva samples were included for study, of which 125 oral squamous carcinoma cases, 124 oral mucosal precancerous lesions and 124 healthy volunteers, all enrolled cases were signed with informed consent. 373 cases were divided into experimental development groups (1)stbatch) and verification group (1)ndbatch), wherein the experimental development group had 193 saliva samples with 65 oral squamous carcinoma cases, 64 oral mucosal precancerous lesions, and 64 healthy volunteers; the test group had 180 cases of oral squamous cell carcinoma, precancerous lesions of the oral mucosa, and 60 cases of healthy volunteers (shown in Table 1).
Table 1 specific clinical data
2. Example two: the experimental development group preliminarily determines the differential metabolic saliva markers among the oral squamous carcinoma group, the oral mucosa precancerous lesion group and the health group.
Method for extracting saliva sample: rinsing the subject with 0.9% normal saline to remove food residues in the mouth, and collecting 1ml of saliva of the subject with a tooth glass after rinsing; then 1200 turns, centrifuges for 5min, collects the supernatant and carries out CPSI-MS detection.
The conductive polymer spray ionization mass spectrometry (CPSI-MS) is an advanced in-situ mass spectrometry technology, and can be used for analyzing lipid molecules in a sample at high flux in a normal pressure and open environment by only using a small amount of sample. Compared with the traditional paper spray ionization, the technology has the following advantages: (1) the sample consumption is low (more than or equal to 100 nL); (2) polar, water-soluble metabolites and drug molecules; (3) the operation is quick and simple; (4) qualitative and semi-quantitative; (5) the sensitivity is high. The characteristics enable the technology to meet the requirement of Point of Care Test (POCT) of clinical samples.
Firstly, 193 saliva samples (65 oral squamous carcinoma cases, 64 oral mucosa precancerous lesions and 64 healthy volunteers) of an experimental development group are subjected to high-throughput metabolic profiling analysis by using a CPSI-MS technology, and 627 m/z ions are obtained in total. Group clustering analysis of the above metabolites using PLS-DA revealed that cases of the same type of group clustered well together (FIG. 1).
Comparative analysis between the two groups (fold change >1.5or <0.667, P <0.05) was performed using student's t test method, resulting in 58 differential metabolites between the precancerous and healthy groups and 106 differential metabolites between the oral squamous carcinoma and precancerous groups (as shown in tables 2 and 3).
TABLE 2 differential metabolites significantly altered in healthy and precancerous groups
*FC represent fold change of PML versus HC,only metabolites with FC values larger than 2.0 or smaller than 0.5 were listed in the table(n＝42).
TABLE 3 differential metabolism with significant alterations in the oral squamous carcinoma and precancerous lesions groups
*FC represent fold change of PML versus HC,only metabolites with FC values larger than 2.0 or smaller than 0.5 were listed in the table(n＝90).
3. Example three: the validation group further validates the differential metabolites determined by the experimental development group.
A further 180 saliva samples, validation groups (oral squamous carcinoma cases, oral mucosal precancerous lesions, 60 in each case in healthy volunteers) were used for validation analysis of 58 and 106 differential metabolites as previously determined. The results show that 42 metabolites of 58 candidate markers determined by the experimental development group oral mucosa precancerous lesion vs healthy volunteers are detected again and have statistical significance (fold change >2.0or <0.5, P < 0.05). The 106 candidate markers identified in the oral squamous carcinoma group vs oral precancerous lesion group were detected again with 94 metabolites and were statistically significant (fold change >2.0or <0.5, P <0.05) (as shown in fig. 2).
4. Example four: the metabolites gradually increased and decreased in the evolution process of healthy volunteers-precancerous lesion-oral squamous cell carcinoma.
Further analysis of the common differential metabolites identified in the experimental development and validation groups identified 8 most significant metabolites with increasing or decreasing trend from healthy volunteers-premalignant-oral squamous cell carcinoma (shown in table 4), with 4 increasing and 4 decreasing metabolites. Statistical analysis proves that the 8 metabolites can effectively distinguish oral mucosa precancerous lesion and oral squamous cell carcinoma patients, and the relative content of the 8 metabolites is gradually increased or decreased from healthy volunteers-precancerous lesion-oral squamous cell carcinoma, and shows the change condition of salivary metabolites in the process of oral squamous cell carcinoma canceration progression. The 8 characteristic metabolites are not only cancerous metabolites for distinguishing oral squamous cell carcinoma patients, but also good markers for predicting the conversion of oral mucosa precancerous lesion to oral squamous cell carcinoma.
TABLE 4 increasing or decreasing metabolites
The structural identification of the target differential metabolite obtained by the statistical analysis is mainly carried out by the following method and flow:
1) further matching and inferring possible molecular structures thereof by online database retrieval;
2) presume the possible molecular formula of metabolite according to the accurate mass number and isotope abundance ratio of the molecular ion peak, mainly obtain [ M + H ] + or [ M-H ] -ion peak of metabolite to the first-level high-resolution mass spectrum;
3) for the different metabolite ions of the commercially available standard, the LC-MS/MS analysis is further performed on the standard of the target metabolite, and the comparison is performed with the cleavage result of the secondary mass spectrum, so as to confirm the structure name of the metabolite.
5. Example five: the salivary metabolism marker combination has the capability of identifying healthy volunteers, precancerous lesion patients and oral squamous carcinoma patients.
When ROC curve analysis is performed on 373 cases by using the 8 markers as a combination, the early healthy volunteers vs precancerous lesion patients (AUC 0.89), the precancerous lesion patients vs oral squamous carcinoma patients (AUC 0.83), and the healthy volunteers vs oral squamous carcinoma patients (AUC 0.96) can be accurately distinguished (see fig. 3, ROC curve graph), and the 8 salivary metabolism marker combinations are proved to be not only the cancerous metabolic markers of oral squamous carcinoma, but also the oral mucosa precancerous lesion patients with high risk of canceration can be effectively screened. The saliva diagnosis kit for oral mucosa precancerous lesion patients (white spots and red spots) is developed based on the marker combination, can realize early, rapid and noninvasive screening of oral mucosa precancerous lesion patients (white spots and red spots) before malignant transformation, and has wide clinical application prospect.
The above description is only a preferred embodiment of the present invention, and should not be construed as limiting the scope of the present invention. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.
Claims (7)
1. A salivary metabolism marker for predicting canceration of a patient with precancerous lesions of oral mucosa, wherein the marker comprises Putrescine Putrescine, Cadaverine Cadaverine, Thymidine Thymidine, Adenosine Adenosine, Hippuric acid, Phosphocholine Phosphocholine, Glucose or arachidonic acid Adrenic acid.
2. The salivary metabolism marker as claimed in claim 1 wherein the marker is: putrescine, Cadaverine, Thymidine and Adenosine are set as a group; the Hippuric acid, Phosphocholine, Glucose and Adrenic acid were set as another group, and the two groups were used in combination.
3. A salivary metabolism marker as claimed in claim 1 or claim 2 wherein the metabolism marker is not only a cancerous metabolism marker of oral squamous cell carcinoma but also a patient with oral mucosal precancerous lesions with high risk of canceration.
4. A kit for predicting the development of canceration in a patient having precancerous lesions of the oral mucosa, said kit comprising a salivary metabolism marker as claimed in any one of claims 1 to 3.
5. The application of the salivary metabolism marker in the preparation of a kit for predicting canceration of a patient with precancerous lesion of oral mucosa.
6. The use according to claim 5, wherein a saliva sample is taken for diagnosis, and the content of Putrescine, Cadaverine, Thymidine, Adenosine is gradually increased from healthy people-patients with precancerous lesions-patients with cancer; the contents of Hippuric acid, Phosphocholine, Glucose and Adrenic acid are gradually decreased from healthy people-precancerous lesion patients-cancer patients.
7. The use of claim 5or 6, wherein the metabolic markers not only diagnose patients with oral squamous carcinoma, but also screen patients with oral mucosal precancerous lesions at high risk of canceration.
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| CN117467765A (en) * | 2023-10-31 | 2024-01-30 | 西南医科大学附属口腔医院 | Application of FGF6 in diagnosis and treatment of oral mucosa canceration |
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| CN117467765A (en) * | 2023-10-31 | 2024-01-30 | 西南医科大学附属口腔医院 | Application of FGF6 in diagnosis and treatment of oral mucosa canceration |
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