Application of Dencichine in preparation of biological medicine preparation for treating lung cancer
A pharmaceutical preparation, biological technology, applied in the direction of drug combination, pharmaceutical formulation, anti-tumor drugs, etc., to achieve the effect of inhibiting lung cancer and inhibiting development
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Examples
Embodiment 1
[0026] Example 1：Dencichine inhibits the tumorigenesis of A549 lung cancer cells in nude mice.
[0027] 1.1 experimental method:
[0028] 1.1.1 nude mice xenotransplantation model verifies the inhibitory effect of Dencichine on the tumorigenesis of lung cancer cells.
[0029] Six-week-old nude mice (BALB / c Nude) were purchased from Changzhou Cavens Experimental Animal Co., Ltd. Lung cancer cell A549 was injected subcutaneously into nude mice, and each nude mouse was injected with one site, and each site was injected with 3×10. 6A cell. After 10 days, xenografts were found under the skin of nude mice. At this time, Dencichine was injected into the tumor again for 15 days, and the nude mice were killed and the tumor was collected. Then, measure the weight of the tumor.
[0030] 1.2 experimental results:
[0031] Dencichine significantly inhibited the tumorigenesis of A549 in nude mice, mainly because the tumor of Dencichine group was smaller and its weight was lower ( Figure 1 A、1B)...
Embodiment 2
[0032] Example 2：Dencichine affects the expression of genes of proliferation, invasion and apoptosis in lung cancer cells
[0033] 2.1 experimental methods:
[0034] 2.1.1 Westernblot detection
[0035] A certain weight of nude mouse xenograft tissue was weighed, and RIPA lysate (RadioImmunoprecipitationAssay) was added to homogenize on ice. Centrifuge for 15min×20000g, and collect the supernatant. Add protein loading buffer and cook the sample in a metal bath at 98℃ for 8 min; Centrifuge for 3min×20000g to remove the precipitate. Electrophoresis with 8% PAGE-SDS gel, loading 30μ g; Transfer the membrane to PVDF membrane, and incubate primary antibody (CyclinA1, N-cadherin, Bax, cleansed caspase-3) and secondary antibody; The target protein band was displayed by developing and photographing, and Tubulin-α was used as internal reference. Image J was used to analyze the gray scale of each protein band, and the gray scale values were counted.
[0036] 2.2 experimental results:
[003...
Embodiment 3
[0038] Example 3：Dencichine affects the proliferation, invasion and apoptosis of A549 cells
[0039] 3.1 Experimental methods:
[0040] 3.1.1 The proliferation of A549 cells was detected by CCK-8 method.
[0041] In this example, CCK-8 kit (Biyuntian, Shanghai, China) was used to detect the growth and proliferation of A549 cells. Will be 3×10 3 Each cell is planted in a 96-well plate. Dencichine was used to incubate A549 cells in the 96-well plate, CCK-8 solution was added every 24 hours for 2 hours, and then the absorbance at OD450 nm was detected by microplate reader. This experiment was repeated three times, and each group of each experiment was repeated with 5 samples.
[0042] 3.1.2 Detection of invasion of A549 cells by trans-well method
[0043] Experiments were carried out in a 24-well plate 8μm Trans-well chamber, which was purchased from Corning (Shanghai, China). First, 1.5×10 of Dencichine-treated and non-dosed samples were added. 5 A549 cells were planted in the upper...
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Description
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Application Information
- IPC
- A61K31/198; A61P35/00
- CPC
- A61K31/198; A61P35/00
- Inventors
- 陈添兴