Application of reagent for inhibiting bio-synthesis of endogenous metabolite hypotaurine in preparation of medicine for inhibiting tumor cell proliferation
A hypotaurine, biosynthesis technology, applied in the medical field, can solve the problems of low annual morbidity and high mortality
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Embodiment 1
[0044] Example 1. Homocysteine inhibits the biosynthesis of intracellular hypotaurine, thereby inhibiting tumor cell proliferation
[0045] Synthetic pathway of hypotaurine in human body, see figure 1 .
[0046] U251 cell line (purchased from KGI Biotechnology, catalog number is KG050, human glioma cells) was used in DMEM high glucose medium (purchased from HYCLONE) containing 10% FBS (purchased from Hangzhou Sijiqing and Harbin Saituo Company) company) at 37°C in a 5% carbon dioxide environment. 250-500 μM homocysteine (HCA) (purchased from Shanghai SIGMA) was added to the medium. HCA can inhibit the synthesis of intracellular hypotaurine. After 24 hours, it can be found that cell proliferation is inhibited by using WST-1 (Beijing ROCHE) staining method to detect the cell mass ( figure 2 ). This inhibition of proliferation was accompanied by a reduction in the amount of intracellular hypotaurine.
[0047] figure 2 It can be seen that with the increasing amount of...
Embodiment 2
[0048] Example 2. Cysteamine promotes the biosynthesis of intracellular hypotaurine, thereby promoting the proliferation of tumor cells.
[0049] Using the U251 cell line, add 200-400 μ M cysteamine (purchased from Shanghai SIGMA) to the medium respectively, and still cultivate it under the conditions described in Example 1, and use the WST-1 staining method to detect the cell mass, and it is found that the cell mass is Proliferation was significantly enhanced. This enhancement was accompanied by an increase in the amount of intracellular hypotaurine ( image 3 ).
[0050] image 3 It can be seen from the data that with the increase of the amount of cysteamine added, the number of cell proliferation increases, and the concentration of intracellular hypotaurine also increases, and this trend is most obvious when the amount of cysteamine added is 200 μM . When the amount of cysteamine added reaches 400 μM, the cell volume is still more than that without adding (proliferates ...
Embodiment 3
[0051] Example 3. Taurine inhibits the biosynthesis of intracellular hypotaurine, thereby inhibiting tumor cell proliferation.
[0052] Using the U251 cell line, add 45-90 μM taurine (purchased from SIGMA) respectively in the culture medium, still cultivate under the conditions described in Example 1, use the WST-1 staining method to detect the cell mass, and find that the proliferation of the cells obviously suppressed. This inhibition was accompanied by a decrease in the amount of intracellular hypotaurine ( Figure 4 ).
[0053] combine Figure 4 It can be seen that the addition of taurine in the medium does lead to an increase in the amount of taurine in the cells, but the amount of hypotaurine is decreasing. This effect was significant over the range of taurine concentrations tested. However, the difference in the inhibitory effect on cell proliferation between 45 μM and 90 μM is not very significant, and factors such as experimental errors cannot be ruled out. But t...
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