Application of phototherapeutic agents in disrupting tumor immunosuppressive microenvironment
An immunosuppressive and microenvironment technology, applied in the field of tumor treatment, can solve the problems of low toxicity and side effects, high cost, achieve low price, improve economic value, and eliminate in situ and metastatic solid tumors.
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Embodiment 1
[0030] The preparation of embodiment 1 cobalt-containing oxide material:
[0031] According to (porous Co 3 o 4 Selective synthesis and characterization of nanosheets and nanorods, Acta Inorganic Chemistry, 2010, 26, 8: 1394-1398) to prepare nano cobalt-containing oxide materials, the specific steps are as follows:
[0032] (1) Dissolve 1 mmol of cobalt chloride and 2 mmol of ammonium fluoride in 24 mL of deionized water, stir for 30 min, then slowly add 2.0 mmol of sodium hydroxide, and continue stirring for 30 min to obtain a mixed solution;
[0033] (2) Transfer the mixed solution in step (1) to a 100mL reactor, react at 120°C for 6h, then cool to room temperature, filter, and wash repeatedly with ethanol and water until neutral, then vacuum-dry at 50°C for 4h ;
[0034] (3) Calcining the dried material in step (2) in a muffle furnace at 400° C. for 2 hours to obtain a cobalt-containing oxide.
[0035] The prepared cobalt-containing oxide was scanned by an electron micr...
Embodiment 2
[0036] Example 2 Validation of cobalt-containing oxides used to activate the immune system to eliminate in situ and metastatic solid tumors
[0037] 4T1 breast cancer cells (purchased from ATCC) were cultured with DEME+10% FBS medium, cultured and passaged, and each mouse was injected with 10 6 4T1 breast cancer cells were injected into both sides of BALB / c mice subcutaneously to establish a model of 4T1 breast cancer cells on both sides of BALB / c mice until the tumor volume was 100 mm. 3 , the tumor-bearing mice were randomly divided into three treatment groups, respectively:
[0038] (1) BALB / c untreated group, that is, tumors on both sides of BALB / c were not treated;
[0039] (2) NSG treatment group, that is, the tumor side of the NSG mice received cobalt-containing oxide + near-infrared light irradiation group, and the other side was not treated;
[0040] (3) BALB / c treatment group, that is, the tumor side of BALB / c mice received cobalt-containing oxide + near-infrared l...
Embodiment 3
[0043] Example 3 Effect of phototherapy on immune microenvironment of tumor tissue
[0044] Collect the tumor tissue of the control group and the BALB / c mouse of the phototherapy group in Example 2 that did not receive the cobalt-containing oxide + near-infrared light irradiation, collect a part of the tumor tissue, and grind it into a single cell suspension solution, cell counting, each take 2×10 7 cells. The reaction system is 100 μL, add CD8-FITC, PD1-PE, TIM3-PE-CY7, CTLA4-APC and LAG3-APC-CY7 according to the volume ratio of 1:50, protect from light, incubate at room temperature for 30 minutes, centrifuge, wash , and then resuspended with 300 μL FCM buffer, and tested on the machine to detect the changes of PD1, TIM3, CTLA4 and LAG3 in CD8+ T cells in tumor tissue. In addition, the same tumor tissue was collected, RNA was extracted by TriZol method, and then RNA reverse transcription was performed with PrimeScript RT kit. Finally, a 15 μL real-time quantitative PCR reac...
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