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Research method for promoting lymphoblast without tumorigenicity to be dedifferentiated in lactic acid/acid environment so as to obtain tumorigenicity

A B-lymphoblastoid and tumorigenic technology, applied in the direction of tumor/cancer cells, blood/immune system cells, biochemical equipment and methods, etc., can solve the problem of unclear lymphoma

Active Publication Date: 2020-04-07
NANTONG UNIVERSITY
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Problems solved by technology

However, the mechanism by which lactic acid induces lymphoma progression has not been elucidated so far
We speculate that lactic acid may be achieved by promoting the expression of key proto-oncogenes in the process of lymphomagenesis, and which of these proto-oncogenes has not been reported so far

Method used

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  • Research method for promoting lymphoblast without tumorigenicity to be dedifferentiated in lactic acid/acid environment so as to obtain tumorigenicity
  • Research method for promoting lymphoblast without tumorigenicity to be dedifferentiated in lactic acid/acid environment so as to obtain tumorigenicity
  • Research method for promoting lymphoblast without tumorigenicity to be dedifferentiated in lactic acid/acid environment so as to obtain tumorigenicity

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Embodiment Construction

[0037] Materials and methods

[0038] 2.1. Materials

[0039]Lactic acid was purchased from Sigma (Shanghai) Company. HMY2-CIR cell line was purchased from ATCC (Manassas, VA). RNase-free DNase I was purchased from Qiagen (Shanghai). TaqTM DNA polymerase was purchased from TaKaRa Biotechnology Co., Ltd. The RevertAidTM First Strand cDNA Synthesis Kit was purchased from Nanjing Novizan Company. Methocult H4230 methylcellulose was purchased from Stem Cell Technologies. PCR primers were ordered from Shanghai Sangon Biotechnology Co., Ltd.

[0040] 2.2. Cell culture

[0041] The culture medium of the HMY2-CIR cell line is 10% fetal calf serum, 100 U / ml penicillin and 100 μg / ml streptomycin added to DMEM high glucose medium, and adjusted to different pH values. The culture condition is 5% CO 2 , 37°C, the medium was changed every three days for eight weeks.

[0042] 2.3. Preparation of low pH medium

[0043] 20 mM lactic acid was dissolved in the above DMEM, the pH value ...

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Abstract

The invention discloses a research method for promoting lymphoblast without tumorigenicity to be dedifferentiated in a lactic acid / acid environment so as to obtain tumorigenicity. The research methodcomprises the steps of: cell culture, low-pH culture medium preparation, cell morphology detection, measurement of cell proliferation, a colony-forming experiment, RT-PCR, data analysis and the like.The method disclosed by the invention is convenient and accurate.

Description

technical field [0001] The invention relates to a research method for lactic acid / acidic environment to promote dedifferentiation of non-tumorigenic lymphoblasts to obtain tumorigenicity. Background technique [0002] Since Otto Warburg put forward the famous "anaerobic glycolysis theory" in 1956, he believed that anaerobic glycolysis played a vital role in the occurrence of tumors, and anaerobic metabolism was also considered as one of the markers of tumors . Even in the presence of oxygen, tumor cells mainly rely on the breakdown of glycogen for energy, resulting in the production and accumulation of high concentrations of lactic acid. Lactic acid does many things. On the one hand, anaerobic glycolysis can generate energy to maintain the growth and other activities of tumor cells; on the other hand, the lactic acid produced by glycolysis can reduce the pH value of the tumor microenvironment, creating a environment that can promote tumor development and metastasis. acidi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/0781C12Q1/6886
CPCC12N5/0693C12Q1/6886C12N2500/60C12N2500/30C12N2506/11C12Q2600/158
Inventor 杨萍胡亚娥
Owner NANTONG UNIVERSITY
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