CN111733236A - PiRNA marker of bile duct cancer and gallbladder cancer and application thereof - Google Patents
PiRNA marker of bile duct cancer and gallbladder cancer and application thereof Download PDFInfo
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- CN111733236A CN111733236A CN202010285175.7A CN202010285175A CN111733236A CN 111733236 A CN111733236 A CN 111733236A CN 202010285175 A CN202010285175 A CN 202010285175A CN 111733236 A CN111733236 A CN 111733236A
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Abstract
The application provides a piRNA marker for diagnosing bile duct cancer and gallbladder cancer and application thereof. The piRNA markers provided by the application comprise one or more of piR-10506469, piR-205488188 and piR-14090389, and can be used for rapidly and accurately screening, predicting, diagnosing or prognosing bile duct cancer or gallbladder cancer.
Description
Technical Field
The application belongs to the field of molecular biology, and particularly relates to a piRNA marker for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer and application thereof.
Background
Exosomes are tiny membrane vesicles secreted by most cells in the body, with lipid bilayer membranes, approximately 30-150nm in diameter. Exosomes are widely existed and distributed in various body fluids, carry and transmit important signal molecules, form a brand-new cell-cell information transmission system, influence the physiological state of cells and are closely related to the occurrence and the progress of various diseases.
Exosomes can directly promote tumor cell growth, invasion and migration by acting as an intercellular communication mechanism. By screening and identifying the small non-coding RNA in the exosome secreted by the tumor cell, a new clue and a more convenient means can be provided for the early diagnosis of the tumor. piRNA is a single-stranded RNA of about 32nt in length, which was first found to be highly expressed in the reproductive system and specifically binds to PIWI protein, regulating gene expression through epigenetic regulation.
Disclosure of Invention
The inventor of the application unexpectedly discovers a piRNA molecular marker which can be accurately and stably used for screening, predicting, diagnosing or prognosing the bile duct cancer and the gallbladder cancer for the first time through exploratory research on piRNA in circulating blood exosomes of patients with the bile duct cancer and the gallbladder cancer.
Based on this, in one aspect, the present application provides a kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer comprising means or reagents for specifically detecting one or more piRNA markers selected from the group consisting of: piR-10506469, piR-205488188, and piR-14090389.
In a specific embodiment, the kit, microarray or biochip provided herein, wherein the means or reagents comprised therein further comprise primers and/or probes capable of specifically detecting said piRNA markers.
In particular embodiments, the primer and/or probe sequences of markers piR-10506469 are: UUGGAUUCCCGGCCAAUGCACCA, respectively; the primer and/or probe sequences for markers piR-205488188 are: GUUCAAUCCCCGGCACCUCCACCA, respectively;
the primer and/or probe sequences for markers piR-14090389 are: GUGUAAACAUCCUACAC UCUCAGCU are provided.
In another aspect, the present application provides the use of a piRNA marker for the manufacture of a kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer, wherein the piRNA marker is selected from one or more of: piR-10506469, piR-205488188, and piR-14090389. In a further aspect, the present application provides the use of a tool or reagent for the specific detection of a piRNA marker in the preparation of a kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer, wherein the piRNA marker is selected from one or more of: piR-10506469, piR-205488188, and piR-14090389.
In a further aspect, the present application provides the use of primers and/or probes for the specific detection of piRNA markers for the preparation of a kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer, wherein the piRNA markers are selected from one or more of: piR-10506469, piR-205488188, and piR-14090389.
In particular embodiments, the piRNA markers to which the present application relates are selected from one or more of the following: initial piR-10506469, precursor piR-10506469, mature piR-10506469, initial piR-205488188, precursor piR-205488188, mature piR-205488188, initial piR-14090389, precursor piR-14090389, and mature piR-14090389.
In particular embodiments, naive piR-10506469, naive piR-205488188, and naive piR-14090389 are capable of being cleaved and expressed intracellularly to the corresponding mature piR-10506469, mature piR-205488188, mature piR-14090389; precursors piR-10506469, piR-205488188, and piR-14090389 can be cleaved and expressed intracellularly to the corresponding mature piR-10506469, mature piR-205488188, mature piR-14090389.
In another specific embodiment, the piRNA markers to which the present application relates are derived from circulating blood exosomes of the subject.
Compared with the existing molecular marker, the piRNA molecular marker provided by the application can accurately screen, predict, diagnose or prognose the occurrence and development of the bile duct cancer and the gallbladder cancer, and has the characteristics of rapidness and stability.
Products or tools, such as kits, microarrays, biochips, or other similar detection tools, prepared based on the piRNA molecular markers provided herein can screen, predict, diagnose, or prognose bile duct cancer or gallbladder cancer by rapid detection of the subject's circulating blood. The method and the related products thereof have high specificity, sensitivity and accuracy.
Drawings
FIG. 1 shows the total expression level of piRNA in circulating blood exosomes of patients with biliary duct cancer and gallbladder cancer.
FIG. 2 shows the expression level of high-level piRNA in circulating blood exosomes of cholangiocarcinoma patients.
FIG. 3 shows the expression level of high-level expression of piRNA in circulating blood exosomes of patients with gallbladder cancer.
FIG. 4 is a diagram of further detecting the expression level of high-expression piRNA in the circulating blood exosomes of patients with cholangiocarcinoma and gallbladder carcinoma by enlarging samples.
FIG. 5 shows the high expression level of piRNA in the circulating blood exosomes after the surgery of the cholangiocarcinoma patient.
FIG. 6 shows the expression level of high-level expression of piRNA in circulating blood exosomes after gallbladder cancer patients' surgery.
Detailed Description
Gallbladder cancer and bile duct cancer are two malignant tumors occurring in the gallbladder, i.e., the gallbladder region, and bile duct cancer, i.e., the tumor occurring in the bile duct. Specific piRNAs for bile duct cancer and gallbladder cancer are rarely reported, and particularly, the identification of piRNAs secreted by cells of the bile duct cancer or the gallbladder cancer into circulating blood exosomes is still blank.
The inventor of the application screens and verifies that the piRNA in circulating blood exosomes is used for predicting the occurrence and the development of the bile duct cancer or the gallbladder cancer for the first time.
Specifically, the inventor of the application unexpectedly discovers a batch of specific high-expression piRNAs in circulating blood exosomes of patients with bile duct cancer and gallbladder cancer through high-throughput sequencing based on samples of Chinese people (mainly Han nationality).
Further, the inventor verifies that the levels of piR-10506469, piR-205488188 and piR-14090389 in the blood exosomes of the bile duct cancer patients are obviously higher than the levels of piR-10506469, piR-205488188 and piR-14090389 in the blood exosomes of normal people through scale-up, and the levels of piR-10506469, piR-205488188 and piR-14090389 in the circulating blood exosomes of the bile duct cancer patients are obviously reduced after operation, so that piR-10506469, piR-205488188 and piR-14090389 in the circulating blood exosomes can be used as markers for screening, predicting, diagnosing or prognosing the bile duct cancer, and a basis is provided for the clinical scheme of preventing or treating the bile duct cancer. For example, if the expression level of any one or more of the circulating blood exosomes piR-10506469, piR-205488188 or piR-14090389 is increased in the subject, the subject may be judged to be at risk of or having bile duct cancer; if the expression level of any one or more of the circulating blood exosomes piR-10506469, piR-205488188 or piR-14090389 of the treated biliary duct cancer patient is reduced, the treatment effect of the biliary duct cancer patient can be judged to be good or the biliary duct cancer patient does not relapse.
In addition, the inventor of the application verifies that the level of piR-10506469 in blood exosomes of gallbladder cancer patients is obviously higher than that of piR-10506469 in blood exosomes of normal people through scale-up, and the level of piR-10506469 in circulating blood exosomes of the gallbladder cancer patients is obviously reduced after operation, so that piR-10506469 in the circulating blood exosomes can be used as a marker for screening, predicting, diagnosing or prognosing the gallbladder cancer, and a basis is provided for the clinical scheme establishment for preventing or treating the gallbladder cancer. For example, if the expression level of circulating blood exosomes piR-10506469 is increased in the subject, the subject may be judged to be at risk of or having gallbladder cancer; if the expression level of the circulating blood exosomes piR-10506469 of the gallbladder cancer patient is reduced after treatment, the gallbladder cancer patient can be judged to have good treatment effect or no relapse.
Based on this, the inventors of the present application have developed products or tools, including kits, microarrays, biochips, or other similar detection tools, that can be used for rapid and accurate screening, prediction, diagnosis, or prognosis of bile duct cancer and gallbladder cancer.
For a better understanding of the present disclosure, reference will now be made to the following detailed description taken in conjunction with the accompanying drawings. It should be understood that these examples are for further illustration only and are not intended to limit the scope of the present application. In addition, after reading the contents of the present application, some insubstantial changes or modifications of the present application will still fall within the scope of the present application.
The experimental procedures in the following examples are, unless otherwise specified, routine in the art.
Example 1
This example provides identification of specific highly expressed piRNAs in circulating blood exosomes of cholangiocarcinoma or gallbladder carcinoma patients by high-throughput sequencing. The specific method is as follows: xue Xingying, et al, Exosomal miRNA profiling before and after surgery with modified cellular diagnostic and cognitive marker for lung adonocardioma, Acta Biochim Biophys Sin (Shanghai), Vol.52, No.3, Pages281-293,2020.
The sampled patient information is shown in table 1.
TABLE 1
The total expression amounts of the piRNA calibrated in the circulating blood exosomes of patients with bile duct cancer and gallbladder cancer are respectively shown in figure 1.
The results indicate that the piRNA highly expressed in circulating blood exosomes of patients with biliary duct cancer comprises: piR-2660989, piR-10506469, piR-20548188, piR-10822895, piR-hsa-23209, piR-18044111, piR-4469033, piR-6756604, piR-4333713 and piR-4494501, wherein the specific data are shown in FIG. 2; the piRNA highly expressed in circulating blood exosomes of patients with gallbladder cancer comprises: piR-2660989, piR-10506469, piR-20548188, piR-10822895, piR-23209, piR-18044111, piR-17603885, piR-5678826, piR-14090389 and piR-32159, and the specific data are shown in FIG. 3.
Example 2
This example further demonstrates the high expression of piRNA in circulating blood exosomes of cholangiocarcinoma and gallbladder carcinoma patients identified in example 1 by expanding the samples.
Specifically, by comparing the levels of piR-10506469, piR-205488188 and piR-14090389 in the blood exosomes of 50 normal persons, 40 bile duct cancers and 25 gallbladder cancer patients, the levels of piR-10506469, piR-205488188 and piR-14090389 in the blood exosomes of the bile duct cancer patients are found to be remarkably higher than the levels of piR-10506469, piR-205488188 and piR-14090389 in the blood exosomes of the normal persons; piR-10506469 in blood exosomes of gallbladder cancer patients were significantly higher than piR-10506469 in blood exosomes of normal persons, as shown in fig. 4.
Example 3
This example further examined the expression level of piRNA in the circulating blood exosomes of patients with biliary duct cancer or gallbladder cancer after receiving surgical treatment.
Specifically, after the patients with bile duct cancer are treated by operation, the expression levels of piR-10506469, piR-205488188 and piR-14090389 in circulating blood exosomes are obviously reduced, as shown in figure 5.
After the patients with gallbladder cancer are treated by operation, the expression level of piR-10506469 in circulating blood exosomes is obviously reduced, as shown in figure 6.
The above description is not intended to limit the present application, nor is the present application limited to the above examples. Those skilled in the art should also realize that such changes, modifications, additions and substitutions are within the spirit and scope of the present application.
Claims (8)
1. A kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer comprising means or reagents for specifically detecting one or more piRNA markers selected from: piR-10506469, piR-205488188, and piR-14090389.
2. The kit, microarray or biochip of claim 1, wherein the means or reagents comprise primers and/or probes capable of specifically detecting the piRNA markers.
3. The kit, microarray or biochip of claim 1 or 2, wherein the piRNA markers are selected from one or more of: initial piR-10506469, precursor piR-10506469, mature piR-10506469, initial piR-205488188, precursor piR-205488188, mature piR-205488188, initial piR-14090389, precursor piR-14090389, and mature piR-14090389.
4. The kit, microarray or biochip of any of claims 1 to 3, wherein the piRNA markers are from circulating blood exosomes of the subject.
Use of a piRNA marker for the manufacture of a kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer, wherein the piRNA marker is selected from one or more of: piR-10506469, piR-205488188, and piR-14090389.
6. Use of a tool or reagent for the specific detection of a piRNA marker in the manufacture of a kit, microarray or biochip for screening, predicting, diagnosing or prognosing bile duct cancer and gallbladder cancer, wherein the piRNA marker is selected from one or more of: piR-10506469, piR-205488188, and piR-14090389.
7. The use of claim 5 or 6, wherein the piRNA marker is selected from one or more of: initial piR-10506469, precursor piR-10506469, mature piR-10506469, initial piR-205488188, precursor piR-205488188, mature piR-205488188, initial piR-14090389, precursor piR-14090389, mature piR-14090389.
8. The use of any one of claims 5 to 7, wherein the piRNA marker is from circulating blood exosomes of the subject.
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| CN114752595A (en) * | 2022-02-19 | 2022-07-15 | 南京鼓楼医院 | Serum tsRNA marker for diagnosing lupus nephritis and screening method and application thereof |
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| CN114752595A (en) * | 2022-02-19 | 2022-07-15 | 南京鼓楼医院 | Serum tsRNA marker for diagnosing lupus nephritis and screening method and application thereof |
| CN114752595B (en) * | 2022-02-19 | 2023-08-01 | 南京鼓楼医院 | Serum tsRNA marker for diagnosing lupus nephritis, screening method and application thereof |
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