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Therapeutic drug for ALT (adult T-cell leukemia) and diagnostic kit

A leukemia and reagent kit technology, applied in the field of biomedicine, can solve problems such as unclear and in-depth research

Active Publication Date: 2019-01-04
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are also related reports on the relationship between ANRIL and tumors, but the research is not in-depth. The molecular mechanism of ANRIL’s carcinogenesis and whether ANRIL can be used as a marker of certain tumor diseases are still unclear.

Method used

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  • Therapeutic drug for ALT (adult T-cell leukemia) and diagnostic kit
  • Therapeutic drug for ALT (adult T-cell leukemia) and diagnostic kit
  • Therapeutic drug for ALT (adult T-cell leukemia) and diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Validation of abnormally high expression of ANRIL in ATL cells

[0026] 1. Cell lines

[0027] ATL cell lines: MT-1, MT-2, MT-4, C8166, ATL-2, ATL-T, TL-Om1 are all from the laboratory of Professor Masao Matsuoka, Kyoto University, Japan.

[0028] Control cell line: Jurkat, from the laboratory of Professor Masao Matsuoka, Kyoto University, Japan.

[0029] 2. Total RNA extraction and reverse transcription

[0030] The specific operations of extraction, purification and detection of total cellular RNA are as follows: 1) Collect a sufficient amount of cells of each strain according to the method of cell passage, transfer to a 1.5mL centrifuge tube without nuclease, centrifuge at 1500rpm for 5 minutes, and discard the supernatant; 2) Add 1mL PBS to each tube to wash the cells twice; 3) Scatter the cells, add 1mL Trizol cell lysate to each tube, and mix well with a pipette to ensure full lysis. Add Trizol cell lysate to the amount of cells); 4) Let stand for 5...

Embodiment 2

[0067] Example 2 Kit for diagnosing adult T-cell leukemia

[0068] Based on the above-mentioned Example 1 and its results, the inventors developed a kit for diagnosing adult T-cell leukemia, which includes reagents for detecting the expression level of ANRIL in adult T lymphocytes.

[0069] The kit of embodiment 2 comprises the primer pair and the reaction system reagent required for the specific detection of the expression level of ANRIL required for the RT-PCR reaction, wherein the sequence of the primer pair required for the RT-PCR reaction is as follows:

[0070] Forward primer sequence: 5'-CAGAGCAATTCCAGTGCAAG-3' (shown in SEQ ID NO.5);

[0071] Reverse primer primer: 5'-GATTTGCAAAAACAGCTG-3' (shown in SEQ ID NO.6).

[0072] RT-PCR reaction system reagents can also contain 2 times PCR Tag enzyme and ddH 2 O. For details of the reaction system of RT-PCR, please refer to Part 3 of Example 1.

[0073] Of course, those skilled in the art can also simply adjust the seque...

Embodiment 3

[0074] Example 3 Kit for diagnosing adult T-cell leukemia

[0075] Based on the above-mentioned Example 1 and its results, the inventors developed a kit for diagnosing adult T-cell leukemia, which includes reagents for detecting the expression level of ANRIL in adult T lymphocytes.

[0076] The kit of Example 3 comprises primer pairs and reaction system reagents for specific detection of the expression level of ANRIL required for the qRT-PCR reaction, wherein the sequences of the primer pairs required for the qRT-PCR reaction are as follows:

[0077] Forward primer sequence: 5'-CAACATCCACCACTGGATCTTAACA-3' (shown in SEQ ID NO.7);

[0078] Reverse primer: 5'-AGCTTCGTATCCCCAATGAGATACA-3' (shown in SEQ ID NO.8).

[0079] The reaction system reagents for qRT-PCR can also include 2 times SYBR enzyme mixture, 50 times ROX reagent and pure water (nuclease-free). For details of the reaction system of qRT-PCR, please refer to Part 4 of Example 1.

[0080] Of course, those skilled ...

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Abstract

The invention relates to a diagnostic kit for ALT (adult T-cell leukemia) and a drug for treating ALT. It is discovered for the first time that detection of the expression level of ANRIL in ATL cellscan become one of diagnostic indexes of ALT and proliferation of the ATL cells can be effectively inhibited and apoptosis of the ATL cells can be effectively promoted through specific interference / knockdown of expression level of ANRIL, and accordingly, the kit for diagnosing ALT and the drug for treating ALT are developed, and a new approach and strategy are provided for diagnosis and treatment of ALT.

Description

technical field [0001] The invention relates to a kit for diagnosing adult T-cell leukemia and a medicine for treating adult T-cell leukemia, belonging to the technical field of biomedicine. Background technique [0002] Adult T-cell leukemia (ATL, adult T-cell leukemia) is a malignant proliferative disease of the lymphoid system. Studies have shown that ATL is caused by human T-cell leukemia virus type 1 (HTLV-1, human T-cell leukemia virus type 1) However, the molecular mechanism of ATL pathogenesis is still not fully elucidated. [0003] Long noncoding RNA (Long noncoding RNA, lncRNA) refers to the noncoding RNA with a length of more than 200 nucleotides that regulates gene expression. More and more studies have shown that lncRNA plays an important role in tumor growth, proliferation, apoptosis and other processes. For example, some lncRNAs bind to DNA, RNA, and protein molecules or interact with their combinations as fundamental regulators in chromatin organization, tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/02
CPCA61K45/00A61P35/02C12Q1/6886C12Q2600/158
Inventor 赵铁军王志龙张熠玲王雍衣凯宁徐玲玲
Owner ZHEJIANG NORMAL UNIVERSITY
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