Application of theanine in preparation of products for preventing and treating cancer diseases
A technology of theanine and inflammation, applied in the medical field, can solve problems such as toxic side effects and limited prevention and treatment
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Embodiment 1
[0020] (1) Example 1: Theanine and its mixture with 6-fluoro-4-hydroxycoumarin inhibit the growth of cancer cells in vitro Inhibition experiment:
[0021] The MTT method was used to test the inhibitory effect of theanine and its mixture with 6-fluoro-4-hydroxycoumarin on the growth of cancer cells in vitro. Proceed as follows:
[0022] 1. Main reagents, cell lines and instruments:
[0023] Human pancreatic cancer PANC-1 and human cervical cancer Hela cell lines and instruments were described above.
[0024] RPMI1640 and DMEM culture medium: Hyclone Company; inactivated fetal bovine serum: Hyclone Company;
[0025] Trypsin (trypsin): Amersco Company; 0.4% trypan blue: Sigma Company; Tetramethylazozolium salt (MTT): Sigma Company;
[0026] 2. Experimental steps:
[0027] (1) Digest the cancer cells in logarithmic growth phase with 0.25% trypsin to make a single cell suspension and adjust the cell concentration to 5×10 4 / mL, inoculated in 96-well culture plate with 100 μl p...
Embodiment 2
[0041](2) Example 2: Theanine and its mixture with coumarin inhibit cancer cell invasion inhibition experiment in vitro:
[0042] 1. Main reagents, cell lines and instruments:
[0043] Human pancreatic cancer PANC-1 and human cervical cancer Hela cell lines and instruments were described above.
[0044] RPMI1640 and DMEM culture medium: Hyclone Company; inactivated fetal bovine serum: Hyclone Company;
[0045] Trypsin (trypsin): Amersco; propidium iodide (PI): Sigma, USA.
[0046] Matrigel, Fibronectin: BD Company of the United States; Transwell Chamber: Costar Company of the United States; cell culture plastic plate (24 wells): BD Company.
[0047] 2. Experimental steps:
[0048] (1) Place the Transwell chamber in a 24-well plate, spread 100 μl / well of Fibronectin and Matrigel evenly on the polycarbonate membrane at the bottom of the chamber, incubate at 37°C for 2 hours, and then air-dry in an ultra-clean workbench;
[0049] (2) Cells were collected, and the upper chambe...
Embodiment 3
[0059] (3) Example 3: The experiment of theanine inhibiting the growth of human cancer xenografts in nude mice
[0060] 1. Experimental animals, cell lines, main reagents and instruments:
[0061] SPF grade BALB / c nude mice, 4-5 weeks old, 18-22g, male, purchased from Beijing Huafukang Experimental Animal Center, animal license number SCXK (Beijing) 2009-0004, bred in SPF grade animal laboratory ; IVC was independently sent back to the air cage, Suzhou Suhang Technology Equipment Co., Ltd.; human pancreatic cancer PANC-1 and human cervical cancer Hela cell lines and other instruments were as described above.
[0062] RPMI1640 and DMEM culture medium: Hyclone Company; inactivated fetal bovine serum: Hyclone Company, stored at -20°C; trypsin: Amersco Company; 0.4% trypan blue: Sigma Company;
[0063] 2. Experimental steps:
[0064] (1) Cell culture and establishment of transplanted tumor animal models: Human pancreatic cancer PANC-1 and human cervical cancer Hela cancer cell l...
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