CN118006780A - As papillary thyroid cancer marker and method of use - Google Patents
As papillary thyroid cancer marker and method of use Download PDFInfo
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Abstract
The invention belongs to the technical field of genetic engineering, and discloses an application method of annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker, which comprises the following steps: collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples; and (3) extracting and detecting the circRNA: separating circRNA from serum and tissue samples by adopting a specific RNA extraction method, and detecting the expression level of hsa_circ_0010023 by using a real-time fluorescent quantitative PCR (RT-qPCR) technology; data analysis: comparing the expression difference of hsa_circ_0010023 in serum of the PTC patient with that of healthy control, and the expression condition of the circRNA in PTC tissue and paracancerous normal tissue; authentication and application: verification studies were performed by expanding the sample size to assess the sensitivity and specificity of hsa_circ_0010023 as a PTC auxiliary diagnostic marker. The invention provides a new means and method for the early diagnosis, prognosis evaluation and treatment monitoring of PTC, and is expected to improve the clinical management and the life quality of PTC patients.
Description
Technical Field
The invention belongs to the technical field of genetic engineering, and particularly relates to an application method of annular RNAhsa _circ_0010023 serving as a papillary thyroid cancer marker.
Background
Papillary Thyroid Carcinoma (PTC) is the most common type of thyroid carcinoma, and early accurate diagnosis is critical to increase patient survival. However, currently available diagnostic methods such as Fine Needle Aspiration Cytology (FNAC) still suffer from certain false negatives and uncertainties. Circular RNA (circRNA) is used as a novel non-coding RNA, is abnormally expressed in various cancers, and is expected to become a novel tumor marker. The prior art has the following technical problems in the early and accurate diagnosis of Papillary Thyroid Carcinoma (PTC):
1. limitations of diagnostic methods
Fine needle puncture cytology (FNAC) is one of the current methods of diagnosing thyroid cancer, but this method has certain limitations. FNAC produces an indeterminate or false negative result in some cases, which leads to inaccurate diagnosis and thus delays treatment. In particular, the accuracy of FNAC can be compromised when the tumor is small or in a location that is difficult to puncture.
2. Lack of specific biomarkers
Although some biomarkers have been used for diagnosis of thyroid cancer, many markers lack sufficient specificity and sensitivity, which limits their use in early diagnosis. In particular, in distinguishing benign from malignant thyroid nodules, existing markers often fail to provide adequate diagnostic information.
3. Diagnostic challenges for early and microscopic lesions
Thyroid cancer, and PTC in particular, appears at an early stage as only tiny lesions that are difficult to detect by routine imaging and FNAC. This poses a challenge for early diagnosis and treatment, as early discovery and treatment are critical in improving patient survival.
4. Requirements for personalized treatment regimens
There are significant biological differences between thyroid cancer patients, which require the identification of specific molecular features during the diagnostic process in order to provide an individualized treatment regimen for each patient. However, existing diagnostic methods often fail to provide adequate molecular level information, limiting the formulation of personalized therapeutic strategies.
5. The potential of non-coding RNAs in cancer diagnosis is underutilized
Recent studies have revealed an important role for non-coding RNAs, particularly circular RNAs (circrnas), in the development of tumorigenesis. These molecules are expected to be novel tumor markers, but the current research on the expression and function of the molecules in thyroid cancer, especially PTC is insufficient, and the potential of the molecules in early diagnosis and treatment monitoring is not fully utilized.
The prior art faces the technical problems of limitations of diagnostic methods, lack of specific biomarkers, early and micro focus diagnosis challenges, individual treatment scheme requirements, underutilization of non-coding RNA potential and the like in the aspect of early and accurate diagnosis of papillary thyroid cancer.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides an application method of annular RNAhsa _circ_0010023 serving as a papillary thyroid cancer marker.
The invention is realized in such a way that a ring RNAhsa _circ_0010023 is used as an application method of a papillary thyroid cancer marker, and the method specifically comprises the following steps:
S1: collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples;
S2: and (3) extracting and detecting the circRNA: separating circRNA from serum and tissue samples by adopting a specific RNA extraction method, and detecting the expression level of hsa_circ_0010023 by using a real-time fluorescent quantitative PCR (RT-qPCR) technology;
S3: data analysis: comparing the expression difference of hsa_circ_0010023 in serum of the PTC patient with that of healthy control, and the expression condition of the circRNA in PTC tissue and paracancerous normal tissue; analyzing the correlation of hsa_circ_0010023 expression level and PTC clinical pathological characteristics (such as tumor size, stage, lymph node metastasis and the like);
s4: authentication and application: performing verification study by enlarging the sample size, and evaluating the sensitivity and the specificity of hsa_circ_0010023 as a PTC auxiliary diagnosis marker; meanwhile, the potential application value of the PTC composition in PTC prognosis evaluation and treatment monitoring is explored.
Further, the S1 specifically includes:
S11: patient recruitment: in cooperation with hospitals, patients diagnosed with Papillary Thyroid Carcinoma (PTC) are recruited and collected as serum samples, PTC tissue samples, and corresponding paracancerous normal tissue samples; at the same time, healthy volunteers were recruited as a control group, and serum samples thereof were collected;
S12: sample processing: after centrifugation, the collected serum sample is split into frozen storage tubes without RNase, and the tissue sample is immediately placed into liquid nitrogen for quick freezing and then transferred to a refrigerator at-80 ℃ for long-term storage; all samples were collected and processed following strict aseptic operating specifications to prevent RNA degradation.
Further, the S2 specifically includes:
(1) RNA extraction: total RNA is extracted from serum and tissue samples by using a special circRNA extraction kit, and the kit can effectively remove linear RNA and enrich the circRNA. The extraction process needs to follow the operation instruction of the kit to ensure the integrity and purity of RNA;
(2) RT-qPCR detection: specific primers for hsa_circ_0010023 were designed and RNA was reverse transcribed into cDNA using reverse transcriptase; then, carrying out PCR amplification by using a real-time fluorescence quantitative PCR instrument, and monitoring the generation amount of a PCR product in real time by detecting the intensity change of a fluorescence signal; finally, the relative expression level of hsa_circ_0010023 is calculated from the standard curve.
Further, the step S3 specifically includes:
(1) Expression differential analysis: using statistical software to conduct differential analysis on the expression level of hsa_circ_0010023 in serum of PTC patients and healthy controls, and comparing the expression differences between the two groups; meanwhile, similar analysis is carried out on the expression conditions in PTC tissues and paracancerous normal tissues;
(2) Correlation analysis: collecting clinical pathology data of PTC patients, including tumor size, stage, lymph node metastasis and other information; then, using correlation analysis methods, the correlation between hsa_circ_0010023 expression levels and these clinical pathology features was studied.
Further, the step S4 specifically includes:
(1) Verification study: in order to verify the reliability of hsa_circ_0010023 as a PTC marker, a verification study is performed by expanding the sample size; serum samples from more PTC patients and healthy controls were collected as described above and tested for hsa_circ_0010023 expression levels; then, its sensitivity and specificity were evaluated by statistical analysis;
(2) Prognosis evaluation and treatment monitoring: collecting preoperative and postoperative serum samples of PTC patients receiving surgical treatment, detecting changes in expression levels of hsa_circ_ 0010023; meanwhile, collecting follow-up information of the patient, including information such as recurrence, metastasis and survival conditions; the potential application value of hsa_circ_0010023 in prognosis evaluation and treatment monitoring is explored by analyzing the correlation between the expression level of hsa_circ_0010023 and the prognosis condition of a patient.
Further, in S1, the serum sample provides a non-invasive pathway to detect circulating RNA (circRNA).
Further, in the step S2, the real-time fluorescent quantitative PCR (RT-qPCR) technology is a high-sensitivity and high-specificity method, and is suitable for quantitatively analyzing the RNA level.
Another object of the present invention is to provide a system for researching the application of the annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker, which implements the method for applying the annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker, the system comprising:
the sample collection module is used for collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples;
The circRNA extraction and detection module is connected with the sample collection module and is used for separating the circRNA from serum and tissue samples by adopting a specific RNA extraction method and detecting the expression level of hsa_circ_0010023 by utilizing a real-time fluorescent quantitative PCR (RT-qPCR) technology;
The data analysis module is connected with the circRNA extraction and detection module and compares the expression difference of hsa_circ_0010023 in serum of the PTC patient and healthy control person and the expression condition of the circRNA in PTC tissue and normal tissue beside the cancer; analyzing the correlation of hsa_circ_0010023 expression level and PTC clinical pathological characteristics (such as tumor size, stage, lymph node metastasis and the like);
The verification and application module is connected with the data analysis module, and is used for carrying out verification research by expanding the sample size and evaluating the sensitivity and the specificity of hsa_circ_0010023 serving as the PTC auxiliary diagnosis marker; meanwhile, the potential application value of the PTC composition in PTC prognosis evaluation and treatment monitoring is explored.
It is a further object of the present invention to provide a computer device comprising a memory and a processor, the memory storing a computer program which, when executed by the processor, causes the processor to perform the steps of the method of applying the annular RNAhsa _circle_ 0010023 as a papillary thyroid cancer marker.
Another object of the present invention is to provide a computer-readable storage medium storing a computer program, which when executed by a processor, causes the processor to execute the steps of the application method of the annular RNAhsa _circle_ 0010023 as a papillary thyroid cancer marker.
In combination with the technical scheme and the technical problems to be solved, the technical scheme to be protected has the following advantages and positive effects:
first, to the technical problems existing in the prior art, some technical effects brought by solving the problems have creative effects. The specific description is as follows:
1. high specificity and high sensitivity detection technology:
Specific primers for hsa_circ_0010023 are developed, and a real-time fluorescent quantitative PCR (RT-qPCR) technology is combined, so that high-specificity and high-sensitivity detection of the circRNA are realized. The technology can accurately distinguish the expression level of hsa_circ_0010023 in the serum of the PTC patient and the healthy control person, and provides a powerful tool for the early diagnosis of PTC.
2. Serum and tissue dual validation system:
A serum and tissue dual verification system was established, not only to detect hsa_circ_0010023 from serum, but also to verify from PTC tissue and paracancerous normal tissue. The system improves the accuracy and reliability of diagnosis and provides more comprehensive diagnosis basis for clinic.
3. Analysis of correlation of clinical pathological features with expression level:
The close relationship between the circRNA and the occurrence and development of PTC is revealed by collecting clinical pathology data of PTC patients and performing correlation analysis with the expression level of hsa_circ_ 0010023. The discovery provides a new thought for researching the pathological mechanism of the PTC and formulating the treatment strategy.
4. Prognosis evaluation and treatment monitoring markers:
preliminary studies have shown that hsa_circ_0010023 has a novel biomarker as a prognosis for PTC and monitoring of therapy. By monitoring the dynamic change of the circRNA in the serum of the patient, the prognosis situation of the patient can be predicted, and a timely and accurate basis is provided for treatment adjustment.
These significant technological advances provide new means and methods for early diagnosis, prognostic evaluation and therapeutic monitoring of PTC, with the hope of improving clinical management and quality of life for PTC patients. In the future, with the deep research and popularization of clinical application, the value of hsa_circ_0010023 as a PTC marker will be widely accepted and applied.
Secondly, the technical scheme is regarded as a whole or from the perspective of products, and the technical scheme to be protected has the following technical effects and advantages:
1. diagnostic effect: the study found that hsa_circ_0010023 was expressed at significantly higher levels in serum and tissues from PTC patients than healthy controls and paracancerous normal tissues, indicating abnormally high expression of the circRNA in PTC. Through ROC curve analysis, hsa_circ_0010023 is found to have higher sensitivity and specificity as a PTC diagnostic marker.
2. Clinical relevance: the expression level of hsa_circ_0010023 is closely related to the clinical pathological characteristics of PTC, such as higher expression level of the circRNA in PTC patients with larger tumor, late stage, and presence of lymph node metastasis. This suggests that hsa_circ_0010023 is involved in the PTC generation and development process.
3. Prognosis evaluation and treatment monitoring: preliminary studies showed that the expression level of hsa_circ_0010023 decreased significantly after PTC patients received surgical treatment and that its dynamic changes correlated with the prognosis of the patient. This suggests that hsa_circ_0010023 has a novel biomarker as a PTC prognostic assessment and therapy monitoring.
The annular RNAhsa _circ_0010023 provided by the invention is abnormally high expressed in papillary thyroid cancer and shows potential application value as a PTC diagnosis, prognosis evaluation and treatment monitoring marker. Future can be further researched and clinically verified to promote the wide application of the PTC battery pack in PTC clinical diagnosis and treatment.
Thirdly, the application method using the circular RNA hsa_circ_0010023 as the Papillary Thyroid Carcinoma (PTC) marker provided by the invention comprises a plurality of steps, and each step represents a remarkable technical progress. In particular, these technological advances are embodied in the following aspects:
1. Targeting and specificity enhancement
Specific circRNA is used as a biomarker hsa_circ_0010023 is used as a research object to specifically identify and distinguish PTC, and the specificity improves the diagnosis accuracy.
Compared to traditional cancer markers, circRNA has higher stability and cell specificity, which enhances the reliability of diagnosis.
2. Non-invasive sample collection
Serum sample analysis the method of extracting circRNA from serum is a non-invasive technique which is safer and more comfortable for the patient than traditional tissue biopsies. The application of this method makes frequent monitoring an advantage for early detection of disease and assessment of treatment progression.
3. Accurate molecular detection technology
Real-time fluorescent quantitative PCR (RT-qPCR) the expression level of hsa_circ_0010023 can be accurately quantified by using RT-qPCR technology, which is a molecular detection technology with very high sensitivity and specificity. Compared to traditional gene expression analysis methods, RT-qPCR provides faster, more accurate data.
4. Comprehensive data analysis and clinical application
And (3) correlation analysis of clinical pathology features, namely correlating the hsa_circ_0010023 expression level with the clinical pathology features of the PTC, and providing important information for typing and prognosis of diseases. The association analysis is helpful for the establishment of personalized treatment schemes and improves the treatment effect.
5. Large-scale validation research and potential applications
And (3) expanding the sample size for verification, namely verifying the sensitivity and the specificity of hsa_circ_0010023 by expanding the sample size, and ensuring the reliability and the universal applicability of the research result. The potential for this approach is not limited to aiding diagnosis, but extends to prognosis evaluation and therapy monitoring.
The method provided by the invention shows remarkable technical progress in the aspect of diagnosing PTC, in particular in the aspects of improving the specificity and accuracy of diagnosis, reducing the burden of patients, providing a more comprehensive disease management scheme and the like. These advances not only promote early diagnosis and effective treatment of PTC, but also help improve the overall therapeutic experience and prognosis of the patient.
Drawings
FIG. 1 is a flowchart of an application method of a ring RNAhsa _circle_ 0010023 as a papillary thyroid cancer marker according to an embodiment of the present invention;
FIG. 2 is a flow chart of a method for collecting samples of serum from PTC patients and healthy controls, and PTC tissue and paracancerous normal tissue samples, according to an embodiment of the present invention;
Fig. 3 is a diagram of an application research system of annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Aiming at the application method of the circular RNA hsa_circ_0010023 as the Papillary Thyroid Carcinoma (PTC) marker, the following two specific embodiments and implementation schemes thereof are provided:
EXAMPLE 1 PTC early diagnosis
Target #: a blood detection method based on hsa_circ_0010023 is developed for early diagnosis of papillary thyroid cancer.
1. Sample collection:
serum samples were collected from suspected PTC patients and healthy controls.
CircRNA extraction and detection:
total RNA, including circRNA, was extracted from serum samples using an RNA extraction kit.
Quantitative analysis was performed against hsa_circ_0010023 using specific RT-qPCR primers and probes.
3. Data analysis:
the difference in hsa_circ_0010023 expression levels in the sera of PTC patients and healthy controls was compared.
Statistical software was used for data analysis to evaluate the correlation of hsa_circ_0010023 expression levels with PTC risk.
4. Clinical application:
Diagnostic criteria are set based on a threshold of hsa_circ_0010023 expression levels.
The method is applied to clinical primary screening and assists in early diagnosis of papillary thyroid cancer.
Expected effect #:
-increasing the early diagnostic rate of PTC.
-Providing a new molecular marker for non-invasive screening.
EXAMPLE 2 PTC treatment effect monitoring
Target #: hsa_circ_0010023 was used as biomarker to monitor the effect of treatment on patients with papillary thyroid cancer.
1. Sample collection:
serum samples were collected before and after PTC patients received treatment.
Circrna detection:
the expression level of hsa_circ_0010023 before and after treatment was detected using a real-time fluorescent quantitative PCR technique.
3. Data analysis and comparison:
The changes in expression levels of hsa_circ_0010023 before and after treatment were compared.
The correlation between changes in hsa_circ_0010023 expression and the therapeutic effect (e.g. tumor shrinkage, reduced biomarker levels, etc.) was analyzed.
4. Clinical application:
hsa_circ_0010023 was used as a biomarker for monitoring treatment response and efficacy.
Personalized treatment regimens and follow-up strategies are provided for the patient.
Expected effect #:
-monitoring the treatment effect in real time and adjusting the treatment regimen in time.
Providing an effective biomarker for the long-term management of PTC patients.
These two examples demonstrate how hsa_circ_0010023 can be used as a biomarker for early diagnosis of PTC and treatment effect monitoring, thereby improving the accuracy of diagnosis and the individuation level of treatment.
As shown in fig. 1, an embodiment of the present invention provides an application method of annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker, which specifically includes:
S1: collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples;
S2: and (3) extracting and detecting the circRNA: separating circRNA from serum and tissue samples by adopting a specific RNA extraction method, and detecting the expression level of hsa_circ_0010023 by using a real-time fluorescent quantitative PCR (RT-qPCR) technology;
S3: data analysis: comparing the expression difference of hsa_circ_0010023 in serum of the PTC patient with that of healthy control, and the expression condition of the circRNA in PTC tissue and paracancerous normal tissue; analyzing the correlation of hsa_circ_0010023 expression level and PTC clinical pathological characteristics (such as tumor size, stage, lymph node metastasis and the like);
s4: authentication and application: performing verification study by enlarging the sample size, and evaluating the sensitivity and the specificity of hsa_circ_0010023 as a PTC auxiliary diagnosis marker; meanwhile, the potential application value of the PTC composition in PTC prognosis evaluation and treatment monitoring is explored.
As shown in fig. 2, the S1 specifically includes:
S11: patient recruitment: in cooperation with hospitals, patients diagnosed with Papillary Thyroid Carcinoma (PTC) are recruited and collected as serum samples, PTC tissue samples, and corresponding paracancerous normal tissue samples; at the same time, healthy volunteers were recruited as a control group, and serum samples thereof were collected;
S12: sample processing: after centrifugation, the collected serum sample is split into frozen storage tubes without RNase, and the tissue sample is immediately placed into liquid nitrogen for quick freezing and then transferred to a refrigerator at-80 ℃ for long-term storage; all samples were collected and processed following strict aseptic operating specifications to prevent RNA degradation.
The step S2 specifically comprises the following steps:
(1) RNA extraction: total RNA is extracted from serum and tissue samples by using a special circRNA extraction kit, and the kit can effectively remove linear RNA and enrich the circRNA. The extraction process needs to follow the operation instruction of the kit to ensure the integrity and purity of RNA;
(2) RT-qPCR detection: specific primers for hsa_circ_0010023 were designed and RNA was reverse transcribed into cDNA using reverse transcriptase; then, carrying out PCR amplification by using a real-time fluorescence quantitative PCR instrument, and monitoring the generation amount of a PCR product in real time by detecting the intensity change of a fluorescence signal; finally, the relative expression level of hsa_circ_0010023 is calculated from the standard curve.
The step S3 specifically comprises the following steps:
(1) Expression differential analysis: using statistical software to conduct differential analysis on the expression level of hsa_circ_0010023 in serum of PTC patients and healthy controls, and comparing the expression differences between the two groups; meanwhile, similar analysis is carried out on the expression conditions in PTC tissues and paracancerous normal tissues;
(2) Correlation analysis: collecting clinical pathology data of PTC patients, including tumor size, stage, lymph node metastasis and other information; then, using correlation analysis methods, the correlation between hsa_circ_0010023 expression levels and these clinical pathology features was studied.
The step S4 specifically comprises the following steps:
(1) Verification study: in order to verify the reliability of hsa_circ_0010023 as a PTC marker, a verification study is performed by expanding the sample size; serum samples from more PTC patients and healthy controls were collected as described above and tested for hsa_circ_0010023 expression levels; then, its sensitivity and specificity were evaluated by statistical analysis;
(2) Prognosis evaluation and treatment monitoring: collecting preoperative and postoperative serum samples of PTC patients receiving surgical treatment, detecting changes in expression levels of hsa_circ_ 0010023; meanwhile, collecting follow-up information of the patient, including information such as recurrence, metastasis and survival conditions; the potential application value of hsa_circ_0010023 in prognosis evaluation and treatment monitoring is explored by analyzing the correlation between the expression level of hsa_circ_0010023 and the prognosis condition of a patient.
In S1, the serum sample provides a non-invasive route to detect circulating RNA (circRNA).
In the step S2, the real-time fluorescence quantitative PCR (RT-qPCR) technology is a high-sensitivity and high-specificity method, and is suitable for quantitatively analyzing the RNA level.
As shown in fig. 3, an embodiment of the present invention provides an application research system for implementing the application method of annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker, wherein the application research system comprises:
the sample collection module is used for collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples;
The circRNA extraction and detection module is connected with the sample collection module and is used for separating the circRNA from serum and tissue samples by adopting a specific RNA extraction method and detecting the expression level of hsa_circ_0010023 by utilizing a real-time fluorescent quantitative PCR (RT-qPCR) technology;
The data analysis module is connected with the circRNA extraction and detection module and compares the expression difference of hsa_circ_0010023 in serum of the PTC patient and healthy control person and the expression condition of the circRNA in PTC tissue and normal tissue beside the cancer; analyzing the correlation of hsa_circ_0010023 expression level and PTC clinical pathological characteristics (such as tumor size, stage, lymph node metastasis and the like);
The verification and application module is connected with the data analysis module, and is used for carrying out verification research by expanding the sample size and evaluating the sensitivity and the specificity of hsa_circ_0010023 serving as the PTC auxiliary diagnosis marker; meanwhile, the potential application value of the PTC composition in PTC prognosis evaluation and treatment monitoring is explored.
An embodiment of the present invention provides a computer device, where the computer device includes a memory and a processor, where the memory stores a computer program, where the computer program, when executed by the processor, causes the processor to execute the steps of the application method of the annular RNAhsa _circle_ 0010023 as a papillary thyroid cancer marker.
An embodiment of the present invention provides a computer-readable storage medium storing a computer program, which when executed by a processor, causes the processor to execute the steps of the application method of the annular RNAhsa _circle_ 0010023 as a papillary thyroid cancer marker.
Example 1: clinical diagnostic study
Implementation scheme #
1. Sample collection and preparation:
Serum samples were collected from thyroid cancer patients and healthy volunteers in hospitals. Ensure that the samples cover patients of different ages, sexes and pathological stages.
And simultaneously collecting tumor tissues and corresponding paracancerous normal tissue samples of the PTC patient.
CircRNA extraction and detection:
The RNA extraction kit was used to extract circRNA from serum and tissue samples.
The expression level of hsa_circ_0010023 was detected using RT-qPCR technique, using specific primers and probes.
3. Data analysis:
Statistical software was used to compare the differences in hsa_circ_0010023 expression in serum of PTC patients versus healthy controls.
The expression of hsa_circ_0010023 in PTC tissue and paracancerous normal tissue was analyzed and its correlation with tumor characteristics was studied.
4. Clinical application exploration:
The potential of hsa_circ_0010023 as a PTC diagnostic marker was assessed from the data obtained.
The application value of the PTC composition in PTC treatment effect monitoring and prognosis evaluation is explored.
Example 2: biomarker mechanism research
Implementation scheme #
1. Biological function study:
In an in vitro cell culture system, the biological function of hsa_circ_0010023 was studied using PTC cell lines and normal thyroid cell lines.
The effect of hsa_circ_0010023 on PTC cell proliferation, migration and invasion capacity was studied by gene knock-down experiments (e.g., using siRNA).
2. Molecular mechanism exploration:
Molecular mechanisms of hsa_circ_0010023 and PTC development were explored using proteomic and transcriptomic methods.
Interactions of hsa_circ_0010023 with other molecules (e.g. miRNA, mRNA) were studied.
3. Clinical sample verification:
A broader clinical sample was used to verify the role of hsa_circ_0010023 in PTC development.
The correlation between hsa_circ_0010023 expression levels and patient prognosis was analyzed.
4. Potential therapeutic target evaluation:
Hsa_circ_0010023 was discussed as a potential therapeutic target, and its potential in targeted therapy PTC was evaluated.
The potential value of hsa_circ_0010023 in thyroid cancer is explored from the angles of clinical application and basic research, and a new view is provided for diagnosis, treatment and mechanism research of thyroid cancer.
The foregoing is merely illustrative of specific embodiments of the present invention, and the scope of the invention is not limited thereto, but any modifications, equivalents, improvements and alternatives falling within the spirit and principles of the present invention will be apparent to those skilled in the art within the scope of the present invention.
Claims (10)
1. A method of using annular RNAhsa _circ_0010023 as a marker for papillary thyroid cancer, the method comprising:
S1: collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples;
S2: and (3) extracting and detecting the circRNA: separating circRNA from serum and tissue samples by adopting a specific RNA extraction method, and detecting the expression level of hsa_circ_0010023 by using a real-time fluorescent quantitative PCR (RT-qPCR) technology;
S3: data analysis: comparing the expression difference of hsa_circ_0010023 in serum of the PTC patient with that of healthy control, and the expression condition of the circRNA in PTC tissue and paracancerous normal tissue; analyzing the correlation of hsa_circ_0010023 expression level and PTC clinical pathological characteristics (such as tumor size, stage, lymph node metastasis and the like);
s4: authentication and application: performing verification study by enlarging the sample size, and evaluating the sensitivity and the specificity of hsa_circ_0010023 as a PTC auxiliary diagnosis marker; meanwhile, the potential application value of the PTC composition in PTC prognosis evaluation and treatment monitoring is explored.
2. The method of using the annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker of claim 1, wherein S1 specifically comprises:
(1) Patient recruitment: in cooperation with hospitals, patients diagnosed with Papillary Thyroid Carcinoma (PTC) are recruited and collected as serum samples, PTC tissue samples, and corresponding paracancerous normal tissue samples; at the same time, healthy volunteers were recruited as a control group, and serum samples thereof were collected;
(2) Sample processing: after centrifugation, the collected serum sample is split into frozen storage tubes without RNase, and the tissue sample is immediately placed into liquid nitrogen for quick freezing and then transferred to a refrigerator at-80 ℃ for long-term storage; all samples were collected and processed following strict aseptic operating specifications to prevent RNA degradation.
3. The method of using the annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker of claim 1, wherein S2 specifically comprises:
(1) RNA extraction: total RNA is extracted from serum and tissue samples by using a special circRNA extraction kit, and the kit can effectively remove linear RNA and enrich the circRNA. The extraction process needs to follow the operation instruction of the kit to ensure the integrity and purity of RNA;
(2) RT-qPCR detection: specific primers for hsa_circ_0010023 were designed and RNA was reverse transcribed into cDNA using reverse transcriptase; then, carrying out PCR amplification by using a real-time fluorescence quantitative PCR instrument, and monitoring the generation amount of a PCR product in real time by detecting the intensity change of a fluorescence signal; finally, the relative expression level of hsa_circ_0010023 is calculated from the standard curve.
4. The method of using the annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker of claim 1, wherein S3 specifically comprises:
(1) Expression differential analysis: using statistical software to conduct differential analysis on the expression level of hsa_circ_0010023 in serum of PTC patients and healthy controls, and comparing the expression differences between the two groups; meanwhile, similar analysis is carried out on the expression conditions in PTC tissues and paracancerous normal tissues;
(2) Correlation analysis: collecting clinical pathology data of PTC patients, including tumor size, stage, lymph node metastasis and other information; then, using correlation analysis methods, the correlation between hsa_circ_0010023 expression levels and these clinical pathology features was studied.
5. The method of using the annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker of claim 1, wherein S4 specifically comprises:
(1) Verification study: in order to verify the reliability of hsa_circ_0010023 as a PTC marker, a verification study is performed by expanding the sample size; serum samples from more PTC patients and healthy controls were collected as described above and tested for hsa_circ_0010023 expression levels; then, its sensitivity and specificity were evaluated by statistical analysis;
(2) Prognosis evaluation and treatment monitoring: collecting preoperative and postoperative serum samples of PTC patients receiving surgical treatment, detecting changes in expression levels of hsa_circ_ 0010023; meanwhile, collecting follow-up information of the patient, including information such as recurrence, metastasis and survival conditions; the potential application value of hsa_circ_0010023 in prognosis evaluation and treatment monitoring is explored by analyzing the correlation between the expression level of hsa_circ_0010023 and the prognosis condition of a patient.
6. The method of using the circular RNAhsa _circ_0010023 as a marker for papillary thyroid cancer according to claim 1, wherein in S1, the serum sample provides a non-invasive route to detect circulating RNA (circRNA).
7. The method of using the circular RNAhsa _circ_0010023 as a marker for papillary thyroid cancer according to claim 1, wherein in S2, the real-time fluorescent quantitative PCR (RT-qPCR) technique is a highly sensitive and highly specific method suitable for quantitative analysis of RNA levels.
8. An application study system for performing the application method of annular RNAhsa _circ_0010023 as a papillary thyroid cancer marker according to any one of claims 1 to 7, wherein the system comprises:
the sample collection module is used for collecting serum samples of PTC patients and healthy controls, and PTC tissues and paracancerous normal tissue samples;
The circRNA extraction and detection module is connected with the sample collection module and is used for separating the circRNA from serum and tissue samples by adopting a specific RNA extraction method and detecting the expression level of hsa_circ_0010023 by utilizing a real-time fluorescent quantitative PCR (RT-qPCR) technology;
The data analysis module is connected with the circRNA extraction and detection module and compares the expression difference of hsa_circ_0010023 in serum of the PTC patient and healthy control person and the expression condition of the circRNA in PTC tissue and normal tissue beside the cancer; analyzing the correlation of hsa_circ_0010023 expression level and PTC clinical pathological characteristics (such as tumor size, stage, lymph node metastasis and the like);
The verification and application module is connected with the data analysis module, and is used for carrying out verification research by expanding the sample size and evaluating the sensitivity and the specificity of hsa_circ_0010023 serving as the PTC auxiliary diagnosis marker; meanwhile, the potential application value of the PTC composition in PTC prognosis evaluation and treatment monitoring is explored.
9. A computer device comprising a memory and a processor, the memory storing a computer program which, when executed by the processor, causes the processor to perform the steps of the method of applying the annular RNAhsa _circ_0010023 as a marker for papillary thyroid cancer as claimed in any one of claims 1 to 7.
10. A computer readable storage medium storing a computer program which, when executed by a processor, causes the processor to perform the steps of the method of applying the circular RNAhsa _circle_ 0010023 of any one of claims 1 to 7 as a marker for papillary thyroid cancer.
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