Phylum: Firmicits strain with anti-cancer activity and anti-cancer uses thereof
A Firmicutes, bacteria technology, applied in bacteria, medical raw materials derived from bacteria, anti-tumor drugs, etc., can solve the problems of destruction, cancer cell drug resistance, physical and mental, powerlessness, etc., and achieve good compliance and remarkable performance. Anticancer activity, low side effects
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Embodiment 1
[0055] Embodiment 1: the screening of CCPM7651 bacterial strain
[0056] 1 Materials and methods
[0057] 1.1 Screening medium and related solutions
[0058] 1.1.1 GRC001 solid medium formula (1L):
[0059] Starch (potato or maize or soybean or millet or other grain or other plant) 20g, KNO 3 1g, K 2 HPO 4 0.5g, MgSO 4 ·7H 2 O 0.5g, NaCl 0.5g, FeSO 4 ·7H 2 O 0.01g, agar 20g; dissolve or suspend starch first, then add other ingredients, remember to shake well after high pressure. Preferably, starch can also be replaced with other examples of suitable carbon sources, including, but not limited to: sugars such as glucose, fructose, sucrose, starch hydrolyzate, cellulose hydrolyzate and molasses; organic acids such as acetic acid, propionic acid , formic, malic, citric, and fumaric acids; and alcohols, such as glycerin. KNO 3 Illustrative examples of other suitable nitrogen sources may also be substituted, including, but not limited to: ammonia, including ammonia gas a...
Embodiment 2
[0071] Example 2: Preparation of bacterial preparations of CCPM7651 strain [i.e. bacterial culture supernatant containing metabolites]
[0072]Experimental bacterial preparation: use the liquid culture obtained from the expanded culture obtained in 2.3 of Example 1, spread plate culture on GRC001 solid medium (GRC001 liquid medium+agar), and then take a single colony from the plate to inoculate 5mL GRC001 In liquid medium, 120 rpm, 14-39°C, culture for 2-14 days, after amplification, centrifuge the bacterial solution at 8000rpm for 10min, and take the supernatant to obtain bacterial preparation (starch medium) for subsequent anti-cancer cell experiments.
[0073] Control bacterial preparation: replace the above-mentioned GRC001 solid medium with LB solid medium (add tryptone, 10 g; yeast extract, 5 g; NaCl, 10 g in 900 mL deionized water. Shake the container to dissolve the solute, add 5 mol / Adjust the pH to 7.0 with L NaOH, then add deionized water to 1 L. Add 15 grams of a...
Embodiment 3
[0074] Embodiment 3: Anticancer activity experiment
[0075] Human ovarian cancer cell line ES2: epithelial ovarian cancer cell line ES2 was purchased from Wuhan Procell Life Science&Technology Co Ltd.
[0076] CCK8: Cell Counting Kit-8 cell counting reagent, Beyotime Biotechnology (Beyotime Biotechnology).
[0077] 3.1ES2 cell culture: plate, 100μl per well, cell density 1x10 5 / mL, after the ES2 cells adhered to the wall, 5 μl of the bacterial preparation product obtained in Example 2 was added thereto, and the control group was an equal volume of GRC001 liquid medium, and then cultivated for 6-24 hours to observe. The morphology of ES2 cells was observed under a microscope.
[0078] The result is as image 3 As shown, the left picture is the ES2 ovarian cancer cells treated with CCPM7651; the right picture is the control treatment; ES2 cells treated with CCPM765 bacterial preparations aggregated, rounded in shape, fell off, and died.
[0079] 3.2 CCK8 test: ES2 cells we...
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