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CDH6-based diagnostic kit for Epstein-Barr virus-associated nasopharyngeal carcinoma

A diagnostic kit and EB virus technology, applied in the field of nasopharyngeal carcinoma diagnostic kits, can solve the problem of lack of research on CDH6, and achieve the effect of great popularization and application value and rapid auxiliary diagnosis.

Active Publication Date: 2019-03-05
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported in the literature that the abnormal expression of CDH6 in kidney and nerve tissue has an important relationship with the prognosis of related cancers such as renal cancer, but there is no research on CDH6 in Epstein-Barr virus-related nasopharyngeal carcinoma

Method used

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  • CDH6-based diagnostic kit for Epstein-Barr virus-associated nasopharyngeal carcinoma
  • CDH6-based diagnostic kit for Epstein-Barr virus-associated nasopharyngeal carcinoma
  • CDH6-based diagnostic kit for Epstein-Barr virus-associated nasopharyngeal carcinoma

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Effect test

Embodiment 1

[0028] Embodiment 1: RT-PCR diagnostic kit for Epstein-Barr virus-associated nasopharyngeal carcinoma of the present invention

[0029] The Epstein-Barr virus-associated nasopharyngeal carcinoma diagnostic kit of the present invention is a real-time quantitative PCR (real-time PCR / RT-PCR) kit, including CDH6-specific primers and internal reference β-actin-specific primers;

[0030] The CDH6-specific primer sequence is:

[0031] Forward: 5'-GTTGCCATCCTTCTGTGCAT-3'

[0032] Reverse: 5'-TTCCTCAGGGTGCCGATATC-3', respectively shown in SEQ ID No.1 and SEQ ID No.2;

[0033] The internal reference gene β-actin primer sequence is:

[0034] Forward: 5'-TCACCAACTGGGACGACATG-3'

[0035] Reverse: 5'-GTCACCGGAGTCCATCACGAT-3', respectively shown in SEQ ID No.3 and SEQ ID No.4;

[0036] The working concentration of the above-mentioned specific primers is 10 pM.

[0037] For more convenient use, this kit also includes an RNA extraction kit and a reverse transcription kit.

Embodiment 2

[0038] Example 2: Using the nasopharyngeal carcinoma diagnostic kit of the present invention to detect nasopharyngeal carcinoma patient tissues and normal human tissues and verify the results

[0039]Five normal human nasopharyngeal tissues and 35 nasopharyngeal carcinoma patient tissues were collected, total RNA was extracted, and real-time fluorescent quantitative PCR was performed after reverse transcription to analyze the differential expression of LMP1, miR-203 and CDH6. Finally, it was found that LMP1 was expressed in almost all tissues, and LMP1 was highly expressed in the tissues of 22 patients with nasopharyngeal carcinoma, compared with normal tissues, P≤0.001, the difference was significant, and the expression of miR-203 was down-regulated, P≤0.001 , the expression of its target gene CDH6 was up-regulated, P≤0.05. Similarly, in 13 NPC patients with low expression of LMP1, the expression of miR-203 was low, P ≤ 0.05, and the expression of target gene CDH6 was increas...

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Abstract

The invention discloses a CDH6-based EBV (Epstein-Barr-virus)-related nasopharyngeal carcinoma diagnosis kit. The kit comprises CDH6 specific primers and internal reference beta-actin specific primers. The CDH6 specific primers are as follows: the sequence of the forward primer is 5'-GTTGCCATCCTTCTGTGCAT-3', and the sequence of the reverse primer is 5'-TTCCTCAGGGTGCCGATATC-3'. The internal reference gene beta-actin primers are as follows: the sequence of the forward primer is 5'-TCACCAACTGGGACGACATG-3', and the sequence of the reverse primer is 5'-GTCACCGGAGTCCATCACGAT-3. The research proves that in the EBV-related nasopharyngeal carcinoma tissues, the detection on the abnormal expression of CDH6 by a realtime-PCR (polymerase chain reaction) technique indicates that the CDH6 can be used as a molecular target for EBV-related nasopharyngeal carcinoma gene diagnosis and therapy like miR-203, and the high expression of the CDH6 can indicate the possibility of EBV-related nasopharyngeal carcinoma infection of the individual to be detected. The nasopharyngeal carcinoma diagnosis kit has the advantages of high simpleness, high speed, high reliability, high specificity and high sensitivity.

Description

technical field [0001] The invention relates to a tumor gene diagnosis kit, in particular to a nasopharyngeal carcinoma diagnosis kit. Background technique [0002] Eptein-Barr virus (EBV) is one of the gamma herpes viruses, which can infect 90% of human beings and transform B lymphocytes in vitro, which is closely related to the occurrence and development of nasopharyngeal carcinoma. Epstein-Barr virus expresses various molecules, such as EBNA1-6, three latent membrane proteins LMP1, 2A and 2B, and miRNAs, at different stages of tumor development. Epstein-Barr virus latent membrane protein 1 (LMP1), the main oncoprotein of Epstein-Barr virus, can induce various signaling pathways, such as NF-KB, MAPK kinase (ERK, p38 and JNK), JAK / STAT and other signaling pathways, causing epithelial cell morphology and Phenotype changes promote epithelial-mesenchymal transition (Epithelial-mesenchymal transition, EMT), which leads to tumor cell invasion and metastasis, and plays an import...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/686C12Q1/6886C12Q2600/158C12Q2561/113C12Q2545/114
Inventor 卢建红左埒莲刘灵芝李桂源张靖
Owner CENT SOUTH UNIV
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