CN109797222B - Liver cancer diagnosis marker, detection primer, kit and application of liver cancer diagnosis marker - Google Patents
Liver cancer diagnosis marker, detection primer, kit and application of liver cancer diagnosis marker Download PDFInfo
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Abstract
The invention belongs to the technical field of liver cancer diagnosis, and discloses a liver cancer diagnosis marker, a detection primer, a kit and application of the marker, wherein the liver cancer diagnosis marker comprises two types of non-coding RNAs (ribonucleic acids) including micro RNAs and lncRNAs, wherein the micro RNAs comprise has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p and has-miR-10b-5p, and the lncRNA comprises lncRNA HOTAIR, lncRNA DANCR, lncRNA HOTTIP and lncRNA TCF 7. The invention has the advantages that the selected marker can play a role in carcinogenesis and cancer suppression, can be used for combined detection and diagnosis of liver cancer, the detection specimen required in the kit is easy to obtain, the clinical operability is strong, the wound is small, the serum microRNAs and lncRNAs have good stability, the detection is convenient, the detection process is simple and convenient, the repetition is easy, the detection can be completed by a common technician, the detection investment of a hospital or a detection mechanism can be greatly reduced, the disease state of a liver cancer patient can be reflected in time, the existing complicated detection is avoided, the time and labor cost is saved, and the individual treatment scheme can be conveniently adopted by a clinician.
Description
Technical Field
The invention belongs to the technical field of liver cancer diagnosis, and particularly relates to a liver cancer diagnosis marker, a detection primer, a kit and application of the liver cancer diagnosis marker.
Background
Primary liver cancer is one of the most common malignant tumors in the world, ranks the fifth most malignant tumor in the world and is also the third most malignant tumor in China. Among them, the most common primary tumor of the liver is hepatocellular carcinoma (HCC). The incidence of primary liver cancer is rising year by year in the world, and the 5-year survival rate of liver cancer patients is less than 10 percent at present, and the primary liver cancer is one of the tumors with the highest malignancy degree. About half of new cases worldwide every year come from China, but in China, most patients have advanced liver cancer when diagnosed, and primary liver cancer also has the characteristics of high incidence and rapid disease progression, and seriously threatens the life health of people. Therefore, the early discovery and early treatment have very important significance for improving the survival rate of the liver cancer patients.
At present, the early diagnosis and screening of liver cancer mainly depend on two means of imaging examination and tumor marker detection. The imaging examination mainly includes Magnetic Resonance Imaging (MRI), CT and ultrasound, and diagnoses the liver by the liver space occupying lesion visible in the visual field, and these means have a certain lag, and the examination depends on the technique and experience of the operator, so that the imaging examination is not suitable for large-scale application, and is not beneficial to the early diagnosis and treatment of liver cancer. Therefore, the detection of tumor markers plays an increasingly important role in the early diagnosis, prognosis, and detection of postoperative recurrence of liver cancer. At present, alpha-fetoprotein (AFP) is still the most common tumor marker for detecting primary liver cancer, but because the sensitivity is low, the specificity is not satisfactory, and misjudgment is easily caused, the search for a new liver cancer diagnosis marker with high specificity, high sensitivity and easy detection has a far-reaching clinical significance.
Micro RNA (micro RNA, miRNA) is a short-chain non-coding RNA containing 18-25 basic groups, and can be combined with a gene or an mRNA specific site to regulate the expression of the gene, thereby playing a wide range of biological roles such as controlling cell growth, proliferation, differentiation, apoptosis, stress and the like. It has been demonstrated that mirnas are resistant to degradation by endogenous enzymes and are therefore stable in plasma, and a large body of research evidence suggests that mirnas play a very important role in the development of cancer. These important studies suggest that mirnas may be biomarkers for tumor diagnosis and targets for prognostic intervention. In the research of liver cancer, some miRNAs including miR-122, miR-26, miR-21, miR-223 and the like are proved to have important cancer inhibiting and cancer promoting effects.
Long Non-coding RNA (lncRNA) with a length of 200-100000nt is a type of RNA without protein coding ability. It is now revealed that lncRNAs can exert many important biological functions, and gene expression can be regulated by exogenous silencing, shear regulation, interaction with miRNAs, protein interaction, genetic variation and other ways. In recent years, more and more lncRNAs related to tumors are discovered, and the lncRNAs become important sources of new tumor markers and drug targets by influencing the growth, apoptosis, infiltration, metastasis and the like of tumor cells. lncRNA HULC, lncRNA PVT1, lncRNA ATB, lncRNA DANCR, lncRNA HEIH and the like are reported to be differentially expressed in primary liver cancer tissues in sequence, are closely related to the occurrence and development of liver cancer, and can be used as biological markers of the liver cancer and reference indexes for prognosis judgment.
The current research suggests that microRNAs and lncRNAs have respective potentials as liver cancer markers, but the following defects still exist: (1) most of researches only select several different microRNAs and lncRNAs as prediction candidate indexes for judging the occurrence and the development of cancers. However, because the index type is single, the liver cancer may not be predicted and diagnosed more accurately, so that the occurrence and development processes of the cancer judged by different markers are different, which greatly interferes the medical workers in judging the course of the patient. (2) Because the types of the indexes are different and the course judgment standards corresponding to each index are inconsistent, fuzzy processing can be carried out on various judgment standards in order to reduce misjudgment of medical workers clinically, so that diagnosis of liver cancer is relatively general, grading prediction diagnosis is not carried out on different stages of liver cancer, accurate judgment of the course of the patient by the medical workers is influenced, and other unnecessary work is increased.
Disclosure of Invention
In order to solve the above problems in the prior art, the present invention aims to provide a diagnostic marker for liver cancer, a detection primer, a kit and use thereof, which can accurately determine the occurrence process of the liver cancer stage.
The technical scheme adopted by the invention is as follows:
the invention provides a liver cancer diagnosis marker, which comprises two types of non-coding RNAs (ribonucleic acids) including microRNAs and lncRNAs, wherein the microRNAs comprise has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p and has-miR-10b-5p, and the lncRNAs comprise lncRNA HOTAIR, lncRNA DANCR, lncRNA HOTTIP and lncRNA TCF 7.
The invention also provides a detection primer of the liver cancer diagnostic marker, the nucleotide sequence of the detection primer is shown in SEQ ID NO. 1-SEQ ID NO.22, wherein SEQ ID NO.1 and SEQ ID NO.2 are used for detecting has-miR-200a-3p, SEQ ID NO.3 and SEQ ID NO.4 are used for detecting has-miR-122-5p, SEQ ID NO.5 and SEQ ID NO.6 are used for detecting has-miR-125-5p, SEQ ID NO.7 and SEQ ID NO.8 are used for detecting has-miR-21-5p, SEQ ID NO.9 and SEQ ID NO.10 are used for detecting has-miR-92b-3p, SEQ ID NO.11 and SEQ ID NO.12 are used for detecting has-miR-10b-5p, SEQ ID NO.13 and SEQ ID NO.14 are used for detecting LncHOIR, SEQ ID NO.15 and SEQ ID NO.16 are used for detecting LncDAR, SEQ ID NO.17 and SEQ ID NO.18 were used for detection of LncRNA HOTTIP, and SEQ ID NO.19 and SEQ ID NO.20 were used for detection of LncRNA TCF 7.
The invention also provides a liver cancer diagnosis kit which comprises a real-time fluorescence quantitative PCR reaction kit and the primer group.
Specifically, the enzyme in the fluorescent quantitative PCR reaction kit is 2 XPCR Master Mix.
Specifically, the kit further comprises a serum RNA extraction kit or a liver tissue RNA extraction kit, and further comprises a reverse transcription kit.
The invention also provides application of the liver cancer diagnostic marker in preparing a kit for predicting a liver cancer occurrence process, wherein the liver cancer markers are lncRNA HOTTIP, hsa-miR-10b-5p and hsa-miR-125b-5p, and the calculation models of the three liver cancer markers are logit (P) ═ 5.214-16.461 miRNA125+30.570 miRNA10b-5208.527 lncTTHOIP.
Preferably, the application is that the detection primer of hsa-miR-125b-5p is shown in SEQ ID NO.7 and SEQ ID NO. 8.
Preferably, the application is that the detection primer of hsa-miR-10b-5p is shown in SEQ ID NO.11 and SEQ ID NO. 12.
Preferably, the application is that the detection primer of lncRNA HOTTIP is shown in SEQ ID NO.17 and SEQ ID NO. 18.
The beneficial effects of the invention are as follows:
(1) the expression quantities of the microRNAs and the lncRNAs in the serum and the tissues of the liver cancer patient are obviously different from the expression quantities of the corresponding tissues of normal people, which shows that the markers can play the roles of carcinogenesis and cancer inhibition, can be used for joint detection and diagnosis of liver cancer, and is particularly used for early diagnosis, prognosis judgment, postoperative recurrence detection and the like of the liver cancer.
(2) The detection specimen in the kit provided by the invention is easy to obtain, strong in clinical operability and small in wound, and serum microRNAs and lncRNAs have good stability and are convenient to detect.
(3) The experimental method related in the kit provided by the invention is mature, the detection process is simple and convenient and is easy to repeat, and the detection can be completed by a common technician, so that the detection investment of a hospital or a detection institution can be greatly reduced.
(4) The kit provided by the invention can reflect the disease state of a liver cancer patient in time, avoid complicated detection, save time and labor cost and facilitate a clinician to adopt an individualized treatment scheme.
Drawings
FIG. 1 shows the expression levels of has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p and has-miR-10b-5p in different liver tissue samples;
FIG. 2 shows the expression levels of IncRNA HOTAIR, IncRNA DANCR, IncRNA HOTTIP, and IncRNA TCF7 in different liver tissue samples;
FIG. 3 is a graph showing the expression levels of has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p and has-miR-10b-5p in different serum samples;
FIG. 4 is a graph showing the expression levels of IncRNA HOTAIR, IncRNA DANCR, IncRNA HOTTIP, and IncRNA TCF7 in different serum samples;
FIG. 5 shows the expression levels of miRNA200a and lncRNA DANCR between HBV positive and HBV negative sera in HCC patients;
FIG. 6 is a ROC (receptor Operating diagnosis) curve for individually detecting ha-miR-200 a-3p, ha-miR-122-5 p, ha-miR-125-5 p, ha-miR-10 b-5p, ha-miR-21-5 p and ha-miR-92 b-3p in serum;
FIG. 7 is ROC curves of separately detecting and diagnosing HCC in serum lncRNA HOTAIR, lncRNA DANCR, lncRNA HOTTIP, and lncRNA TCF7, respectively;
FIG. 8 is a ROC curve for the diagnosis of HCC in combination with the HAs-miR-125-5p, HAs-miR-10b-5p and lncRNA HOTTIP detection in serum.
Detailed Description
The invention is further explained by combining the drawings and specific test examples.
For convenience of mapping, the has-miR-200a, has-miR-122, has-miR-125, has-miR-21-5p, has-miR-92b, and has-miR-10b in FIGS. 1, 3, and 5 correspond to has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p, and has-miR-10b-5p, respectively, hereinafter.
Test example 1: collection and preparation of tissue specimens
Grouping: liver cancer patients (HCC) and hepatic hemangioma patients (a common liver benign tumor) are collected, the hepatic hemangioma patients are used as liver tissue specimens of a healthy control group, the liver tissue specimens are grouped according to the grouping standard shown in table 1, and the liver cancer and the control group specimens thereof are set according to the principle of gender and age matching.
Collecting hepatic hemangioma tissues and liver cancer tissues excised from hepatobiliary surgery in general hospitals in western war zone of people's liberation military in China, wherein the liver cancer tissues are respectively sampled from cancer tissues and tissues beside cancer. The samples are wrapped with ice cubes after being collected and stored in a refrigerator at the temperature of 80 ℃ below zero within 15min for later use.
Test example 2: collection and preparation of serum samples
Grouping: blood samples of healthy persons (HC, patients with hepatic hemangioma) and patients with liver cancer (HCC) were collected, and grouped according to the grouping standards shown in Table 1, and liver cancer and its control group samples were set according to the principle of gender and age-matching. The clinical pathological characteristics of liver cancer patients and healthy patients are shown in table 2.
TABLE 1 grouping criteria for participating populations
TABLE 2 clinical pathological characteristics of liver cancer patients and healthy persons
Collecting samples: 4ml of peripheral venous blood before operation of the liver cancer patient and 4ml of peripheral venous blood of a healthy person are extracted by using a dry blood collection tube and are kept still at 4 ℃ for more than half an hour. Centrifuging the peripheral venous blood at 4 deg.C 400g for 10min, collecting supernatant, centrifuging the supernatant at 4 deg.C 1800g for 10min, collecting supernatant to obtain serum, subpackaging, and storing at-80 deg.C for use.
Test example 3: extraction of RNA from tissue
Extraction was performed with reference to kit instructions. Adding 1ml TRIZOL reagent into every 50-100mg tissue sample, homogenizing with electric homogenizer, and incubating for 5 min; adding chloroform, adding 0.2ml chloroform to 1ml TRIZOL reagent homogenate sample, shaking tube vigorously, incubating for 2-3min, and centrifuging at 4 deg.C for 15min at 12,000 g; transferring the water phase obtained after centrifugation into a new centrifuge tube, adding isopropanol, uniformly mixing, incubating for 10min, and centrifuging for 10 min; removing the supernatant, adding 75% ethanol (at least 1ml of 75% ethanol is added to each 1ml of TRIZOL reagent homogenate sample), centrifuging at 4 deg.C for 5min at 7,500g, and discarding the supernatant to obtain RNA precipitate; drying RNA precipitate in air (in sterile workbench) for 5-10min, adding water without RNase, repeatedly blowing with gun for several times, incubating at 55-60 deg.C for 10min, dissolving RNA precipitate to obtain RNA solution, and storing at-70 deg.C.
Test example 4: extraction of RNA from serum
Extraction was performed with reference to kit instructions. Mixing 200 μ l of blood sample, 750 μ l of TRI REAGENT BD reagent and 20 μ l of glacial acetic acid, shaking vigorously and mixing well, and incubating for 5 min; adding 0.2ml chloroform, shaking the tube manually and vigorously, incubating for 2-3min, and centrifuging at 4 deg.C for 15min at 12,000 g; transferring the water phase obtained after centrifugation into a new centrifuge tube, adding 500 μ l isopropanol, mixing uniformly, incubating for 10min, and centrifuging at 4 deg.C for 10min at 12,000 g; removing the supernatant, adding 75% ethanol (at least 1ml of 75% ethanol is added to 750. mu.l of TRI REAGENT BD reagent homogenate sample), shaking, centrifuging at 4 deg.C for 5min at 7,500g, and discarding the supernatant to obtain RNA precipitate; drying RNA precipitate in air (in sterile workbench) for 5-10min, adding water without RNase, repeatedly blowing with gun for several times, incubating at 55-60 deg.C for 10min, dissolving RNA precipitate to obtain RNA solution, and storing at-70 deg.C.
Test example 5: reverse transcription of serum and tissue total RNA
The RNAs obtained in test examples 3 and 4 were reverse-transcribed using a reverse transcription kit, respectively, and the reverse transcription system and conditions were as described in the specification, and the obtained cDNA was stored at-20 ℃ for use. The reverse transcription kit should be provided with a universal downstream primer for carrying out the next real-time quantitative fluorescent PCR reaction.
Test example 6: quantitative polymerase chain reaction
1. All the cDNA samples obtained in test example 5 were prepared into a Realtime PCR reaction system. The premixed liquid in the system is prepared as follows:
TABLE 3
After the premix was prepared, the solution was mixed by flicking the bottom of the tube and centrifuged briefly at 5000 rpm. The primer F in Table 3 is a primer labeled GSP or F in Table 4, and the primer R in Table 3 is a primer corresponding to the GSP or R in Table 4 and labeled R.
2. Sample adding: mu.l of the mixture was added to each well of the 384-PCR plate, and 2. mu.l of the corresponding cDNA was added to each well. Carefully stick the Sealing Film on the Sealing Film, and mix by centrifugation for a short time. The loaded PCR plate was placed on ice before setting up the PCR program.
3. Reaction: the 384-PCR plate was placed on a Realtime PCR machine for qPCR reaction. The reaction conditions are as follows: at 95 ℃ for 10 min; 40 PCR cycles (95 ℃ C., 10 sec; 60 ℃ C., 60 sec (fluorescence collection)).
TABLE 4 primers used for qPCR
Test example 7: detection of expression levels of miRNAs and lncRNAs in liver tissue
The expression levels of has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p, has-miR-10b-5p, LncRNA HOTAIR, LncRNA DANCR, LncRNA HOTTIP, and LncRNA TCF7 were extracted and examined from selected samples of liver cancer tissue, paracarcinoma tissue, and hepatic hemangioma tissue using the methods described in test examples 1, 3, 5, and 6, and the results are shown in FIGS. 1 and 2. In fig. 1, the absence of "+" indicates that the difference is not significant (P >0.05), "+" indicates that the difference between the two is significant (P <0.01), and the numbers of the presence of "+" and "+" in fig. 2 to 5 are the same as those in fig. 1.
As can be seen from FIG. 1, the expression level differences of has-miR-200a-3P, has-miR-125-5P and has-miR-92b-3P in hepatic hemangioma tissues, paracancerous tissues and liver cancer tissues are not significant (P > 0.05); the expression level of has-miR-122-5P in the liver cancer tissue is extremely lower than that of the adjacent tissues (P <0.01), and the expression level of has-miR-10b-5P in the liver cancer tissue is significantly higher than that of the adjacent tissues (P < 0.05). And the expression level of has-miR-21-5P in liver cancer tissues is remarkably higher than that in liver hemangioma tissues and paracarcinoma tissues (P < 0.01). Thus, has-miR-122-5p is verified to be a cancer suppressor in liver cancer; has-miR-10b-5p and has-miR-21-5p are carcinogenic factors, and lay a good foundation for screening of subsequent tumor biomarkers.
As can be seen from FIG. 2, the expression amounts of LncRNA HOTAIR and LncRNA HOTTIP were not significantly different in hepatic hemangioma tissue, paracarcinoma tissue and liver cancer tissue (P > 0.05); the expression level of LncRNA DANCR in liver cancer tissue is obviously higher than that in liver hemangioma tissue (P <0.05), and is extremely higher than that in para-cancer tissue (P < 0.01); in addition, the expression level of LncRNA TCF7 in liver hemangioma tissues is significantly higher than that in paracarcinoma tissues (P <0.05), and is very significantly higher than that in paracarcinoma tissues (P < 0.01). It can be seen that LncRNA DANCR and LncRNA TCF7 are indeed associated with the malignancy of the tumor.
Test example 8: detection of expression levels of miRNAs and lncRNAs in serum samples
The expression levels of has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p, has-miR-10b-5p, LncRNA HOTAIR, LncRNA DANCR, LncRNA HOTTIP and LncRNA TCF7 were measured from serum samples of selected liver cancer groups (i.e., the liver cancer serogroup shown in FIG. 4) and healthy human groups (i.e., the hepatic hemangioma serogroup shown in FIG. 4) by the methods of test examples 2, 4, 5 and 6, and the results are shown in FIGS. 3 and 4.
As shown in FIG. 3, the expression level difference of has-miR-200a-3P in serum of hepatic hemangioma and liver cancer is not significant (P is more than 0.05); the expression level of has-miR-10b-5P and has-miR-21-5P in liver cancer serum is significantly higher than that in liver hemangioma serum (P is less than 0.01); the expression level of has-miR-92b-3P in liver cancer serum is obviously higher than that in liver hemangioma serum (P is less than 0.05); the expression level of has-miR-122-5P in liver cancer serum is obviously lower than that in liver hemangioma serum (P < 0.05). The expression level of has-miR-122-5P in liver cancer serum is very obviously lower than that in liver hemangioma serum (P < 0.01). The expression result of serum is similar to that of tissue, so that different non-coding RNAs have different functions in hepatocellular carcinoma, and can be used as a marker for early diagnosis and treatment of hepatocellular carcinoma.
As can be seen from FIG. 4, the difference in expression level between LncRNA HOTAIR and LncRNA TCF7 in serum of hepatic hemangioma and liver cancer was not significant (P > 0.05); the expression level of LncRNA DANCR and LncRNA HOTTIP in the serum of hepatic hemangioma is significantly higher than that of LncRNA DANCR and LncRNA HOTTIP in the serum of liver cancer (P < 0.01).
Test example 9: expression levels of both hepatitis B positive and hepatitis B negative in different non-coding RNAs in liver cancer patients.
The positive and negative hepatitis B in the selected group of liver cancers were examined, and the expression levels of serum has-miR-200a-3p, has-miR-122-5p, has-miR-125-5p, has-miR-21-5p, has-miR-92b-3p, has-miR-10b-5p, LncRNA HOTAIR, LncRNA DANCR, LncRNA HOTTIP, and LncRNA TCF7 were extracted and examined for two groups of patients by the methods described in test examples 1, 3, 5, and 6, and the results are shown in FIG. 5.
As can be seen from FIG. 5, the expression level of has-miR-200a-3P and LncRNA DANCR in hepatitis B positive (HBV-related HCC) patients is significantly higher than that in hepatitis B negative (Non-HBV-related HCC) (P < 0.01). The remaining lncRNAs tested did not significantly differ from whether or not hepatitis B was positive in hepatocellular carcinoma (not shown in FIG. 5).
Test example 10: predictive diagnostic effect of non-coding RNA combination on liver cancer
Firstly, carrying out statistical analysis on data, describing quantitative data distribution conditions by adopting a mean plus or minus standard deviation, comparing differences among groups by adopting independent sample t test, then constructing a prediction model by adopting multi-factor stepwise logistic regression analysis, and finally analyzing the prediction effect of the prediction model by adopting an ROC curve. Through statistical analysis, a model for diagnosing liver cancer by logistic regression prediction is established, and logic (P) ═ 5.214-16.461 miRNA125+30.570 miRNA10b-5208.527 lnc HOTTIP. And respectively constructing ROC curves of ten non-coding RNAs and ROC curves for predicting and diagnosing combined miRNA125, miRNA10b and lnc HOTTIP. The ROC curves of six serum mirnas for HCC diagnosis alone are shown in fig. 6, the ROC curves of four lncrnas for HCC diagnosis alone are shown in fig. 7, and the ROC curves of miRNA125, miRNA10b and lnc HOTTIP for HCC diagnosis in combination are shown in fig. 8.
The larger the area AUC value under the ROC curve, the greater the diagnostic value for the disease. Comparing fig. 6 and fig. 7, it can be seen that the AUC value in fig. 8 is higher than the AUC value of any single ncRNA ROC curve, i.e. the AUC value of the combination of the three is greater than the AUC for diagnosing liver cancer alone, so the effect of diagnosing liver cancer by the combination of the three is the best.
The combination of the microRNA and the lncRNA has good prediction effect in early diagnosis of liver cancer, which is shown in the following steps: first, the combined ROC area under the curve (AUC) is larger, indicating that it has good performance. Secondly, the combination comprises microRNA and lncRNA, and the marker types are more comprehensive and novel. Finally, the combined marker contains carcinogenic and cancer suppressor factors at the same time, and can be more comprehensively and objectively used for early prediction and diagnosis of hepatocellular carcinoma; and the markers adopted in the combination are closely related to tumor stem cells and the tumor EMT process, so that the reliability of the tumor stem cells and the tumor EMT process can be further explained.
Test example 11: production of non-coding RNA kit
The kit is used for prediction, early warning and diagnosis of liver cancer, in particular early liver cancer, and consists of a serum RNA extraction system, a reverse transcription system, a real-time fluorescent quantitative PCR system, a primer system and a logistic regression analysis method for evaluating whether liver cancer is suffered. The serum RNA extraction system, the reverse transcription system and the real-time fluorescence quantitative PCR system are respectively a serum RNA extraction kit, a reverse transcription kit (reverse transcription kit for real-time fluorescence quantification) and a real-time fluorescence quantitative PCR reaction reagent which are currently available on the market. The larger the area AUC value under the ROC curve, the greater the diagnostic value for the disease.
The present invention is not limited to the above-described alternative embodiments, and various other forms of products can be obtained by anyone in light of the present invention. The above detailed description should not be taken as limiting the scope of the invention, which is defined in the claims, and which the description is intended to be interpreted accordingly.
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Claims (1)
1. Use of a liver cancer diagnostic marker for preparing a kit for diagnosing liver cancer, wherein the liver cancer marker consists of lncRNA HOTTIP, hsa-miR-10b-5p and hsa-miR-125b-5p, and the calculation model of the three liver cancer markers is logit (P) =5.214-16.461 miRNA125+30.570 miRNA 10-10 b-5208.527 lncHOTTIP; sample-derived serum; wherein, the detection primer of the hsa-miR-125b-5p is shown in SEQ ID NO.5 and SEQ ID NO. 6; the detection primers of the hsa-miR-10b-5p are shown in SEQ ID NO.11 and SEQ ID NO. 12; the detection primers of the lncRNA HOTTIP are shown in SEQ ID NO.17 and SEQ ID NO. 18.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201910203973.8A CN109797222B (en) | 2019-03-18 | 2019-03-18 | Liver cancer diagnosis marker, detection primer, kit and application of liver cancer diagnosis marker |
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| CN114134147B (en) * | 2021-11-16 | 2023-11-14 | 山东大学 | Non-coding RNA for regulating FZD9 and application thereof |
| CN114836538B (en) * | 2022-04-14 | 2023-04-07 | 南昌大学第一附属医院 | Application of biomarker in diagnosis and prognosis of HBV (hepatitis B virus) -related liver cancer |
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