Potentiator of antitumor activity of chemotherapeutic agent
A chemotherapeutic agent and anti-tumor active technology, applied in the field of production or treatment, can solve the problems that have not yet clarified the influence of branched-chain amino acid differentiation, and achieve the effect of preventing metastasis and recurrence
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[0106] An embodiment wherein only isoleucine or a salt thereof is contained as branched-chain amino acid;
[0107] An embodiment wherein only leucine or a salt thereof is contained as branched chain amino acid;
[0108] An embodiment wherein only valine or a salt thereof is contained as branched chain amino acid;
[0109] An embodiment comprising isoleucine or a salt thereof and leucine or a salt thereof as branched-chain amino acids;
[0110] An embodiment comprising isoleucine or a salt thereof and valine or a salt thereof as branched-chain amino acids;
[0111] An embodiment comprising leucine or a salt thereof and valine or a salt thereof as branched-chain amino acids; and
[0112] An embodiment wherein isoleucine or a salt thereof, leucine or a salt thereof, and valine or a salt thereof are contained as branched-chain amino acids.
[0113] The agent or composition of the present invention preferably contains isoleucine or a salt thereof, leucine or a salt thereof, and ...
Embodiment 1
[0171] Example 1: Effect of BCAA on the expression of stem cell markers and differentiation markers in the human liver cancer cell line HAK4
[0172] Human hepatoma cell line HAK4 cells were plated at a concentration of 12,000 cells / well and cultured in 2 mM BCAA or LC 2.5% medium for 72 hours. BCAA is a mixture of isoleucine, leucine and valine (weight ratio: isoleucine:leucine:valine=1:2:1.2). The concentration of BCAA "2 mM" is the total concentration of all amino acids of isoleucine, leucine and valine.
[0173] The composition of LC 2.5% medium is shown in the table below. It should be noted that the "2.5%" in LC 2.5% is the bovine serum content.
[0174] Table 1
[0175] Composition (nmol / ml) Glycine 280 L-alanine 307 L-serine 151 L-threonine 138 L-cystine 2HCl 67 L-Methionine 75 L-Glutamine 689 L-Asparagine 64 L-glutamic acid 53 L-Aspartic Acid 4 L-valine 175 L-leucine 100 L-i...
Embodiment 2
[0187] Example 2: Effect of the combined use of 5FU (5-fluorouracil) and BCAA on the human liver cancer cell line HAK4 (in vitro test)
[0188] At a concentration of 3000 cells / well, the human liver cancer cell line HAK4 cells were plated and cultured for 72 hours under the following conditions:
[0189] ?LC 2.5% Medium
[0190] ?BCAA (2 mM)
[0191] ?5FU (2.5 μg / ml)
[0192] • BCAA (2 mM) + 5FU (2.5 μg / ml).
[0193] After 72 hours, cells were stained with Annexin V using the ApoAlert Annexin V-FITC Apoptosis Kit (Clontech), fixed, and apoptotic cells (%) were detected by Array Scan (Thermo Fisher) .
[0194] Specifically, cells in a 96-well plate were washed with 1X binding buffer, 40 μl of 1X binding buffer containing annexin V (1 μl) was added to each well, and cells were incubated in the dark at room temperature for 5 –15 min. Cells were washed and fixed with 2% formaldehyde. Add 100 μl of PBS containing Hoechst33258 solution, and incubate the cells in the dark fo...
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