Method and kit for treating cancer through combination therapy
A technology of cancer and therapy, applied in the field of cancer treatment through combination therapy, which can solve problems such as drug-resistant diseases
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Embodiment 1
[0049] Preparation of polysaccharide extract and PD-1 inhibitor
[0050] Traditional Chinese medicines containing dendrobii caulis (Dendrobii caulis), ginseng (Panax Gingseng), astragalus, poria cocos, etc. are ground, immersed in deionized water, and extracted using boiling water. Especially, 200g of various traditional Chinese medicines are refluxed with 8-10 times of water (1600-2000ml) for 2 hours. Then repeat the extraction. After extraction, the mixture was left overnight at room temperature, or the mixture was centrifuged below 3000 g for 30 min. The supernatant was subsequently collected and the precipitate was concentrated with 100% ethanol at a volume ratio of 1:4. The residue was further treated with Sevag reagent to remove protein and the supernatant fraction was loaded on a DEAE-cellulose column. The neutral polysaccharide fraction was eluted with water solvent, while the acidic polysaccharide fraction was eluted with 0.5M sodium chloride. Next, these fraction...
Embodiment 2
[0053] Antitumor effects on tumor growth in animal models
[0054] Establish animal models, especially LLC tumor models, to evaluate the anti-tumor effect of the combination therapy of the present invention (ie, polysaccharide extract and PD-1 inhibitor). Animal studies were performed according to institutional guidelines. LLC cells (2x10 6 ), to construct an animal model, that is, C57BL / 6 mouse Lewis lung cancer model (LLC). On day 7, mice bearing LLC tumors with similar tumor size were randomly divided into four groups (n=4), these groups were control group (i.e. PBS), anti-PD-1 antibody (10 mg / kg) (RMP1 -14, Bio X Cell, each injection 250 μ g), the polysaccharide extract (200 mg / kg) in Example 1, or the combination of anti-PD-1 antibody and polysaccharide extract were processed. Specifically, the anti-PD-1 antibody was injected intraperitoneally every 3 days (that is, on days 3, 6, 9, 12, and 15). The polysaccharide extract was intragastrically administered to mice ever...
Embodiment 3
[0058] CD4 + and CD8 + The effect of T lymphocyte activity
[0059] The inventors further assessed whether the combination therapy of the present invention can enhance CD8 + and CD4 + Recruitment and activation of T lymphocyte infiltration. On the 18th day, the mice in each treatment group and control group were sacrificed, and the tumor tissues were collected and fixed with 10% formalin. After fixation, the tissues were embedded in paraffin to prepare slides with tissue sections (5 μm). Sections were deparaffinized in xylene and dehydrated through a series of alcohol fractions. Then, stain with Mayer's hematoxylin (Mayer's hematoxylin, Sigma-Aldrich) for 2 minutes, stain with 0.1% sodium bicarbonate blue for 1 minute, wash with water, and counterstain with Eosin Y solution (Sigma-Aldrich) for 1 minute. Subsequently, antigen retrieval was performed using Novocastra Epitope Retrieval Solution pH 6.0 for 30 minutes in PT Link Dako pretreatment module at 97°C. Slides were ...
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