Application of Cryptotanshinone in Preparation of Tumor Medicine
A cryptotanshinone, experimental technology, applied in the application field of cryptotanshinone in the preparation of tumor drugs, can solve the problem of not many studies on the mechanism of action, and achieve the effect of increasing phosphatase activity
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Embodiment 1I
[0021] Example 1 IGF-1 induces cell proliferation
[0022] Add final concentrations of 1, 3, 10, 30, 100 μg·L –1 IGF-1 treated PC12 cells and hRPE cells. 10 μL of IGF-1 was added to each well, and the same volume of DMEM medium was added to the control group. 3-5 replicate wells were set up for each concentration. Placed at 37°C, 5% CO 2 After culturing in the cell culture box for 24 h, the medium was aspirated, and MTT assay was performed to determine the effect of IGF-1 on the proliferation of PC12 cells and hRPE cells.
[0023] MTT test results such as figure 1 and figure 2 shown, from figure 1 It can be seen from the figure that after different concentrations of IGF-1 act on PC12 cells, compared with the blank group, IGF-1 can significantly promote cell proliferation in a dose-dependent manner. The concentration of IGF-1 was 3 μg·L –1 has promoted the proliferation of PC12 cells ( P –1 , the promoting effect on cell proliferation has been extremely significant ( ...
Embodiment 2
[0025] Example 2 Inhibitory effect of cryptotanshinone on cell proliferation induced by IGF-1
[0026] In order to investigate whether CTS can inhibit the proliferation of PC12 cells induced by IGF-1, we first added CTS (20 μM) to pretreat PC12 cells for 40 min, and then added IGF-1 (10 μg / L) to act on PC12 cells for 24 h, and MTT The effect of CTS on IGF-1-induced cell proliferation was detected by the method. At the same time set only 10 μg·L –1 A control group treated with IGF-1; a control group treated with CTS only (20 μM); and a control group cultured without IGF-1 and CTS. 3-5 replicate wells were set up for each concentration. Placed at 37°C, 5% CO 2 After culturing in the cell incubator for 24 h, the medium was sucked off, and MTT assay was performed to investigate the effect of CTS on IGF-1-induced cell proliferation.
[0027] MTT test results such as image 3 As shown, it can be seen that compared with the blank control group, the IGF-1 treatment group has a s...
Embodiment 3
[0028] Example 3 Inhibitory effect of cryptotanshinone on IGF-1R signaling pathway
[0029] 1) The effect of CTS on the phosphorylation levels of IGF-1R, PI3K / Akt and Ras / Erk proteins induced by IGF-1
[0030] In order to study the effect of CTS on the phosphorylation of IGF-1R and its downstream PI3K / Akt and Ras / Erk induced by IGF-1 stimulation, we intervened PC12 cells with CTS, followed by IGF-1 treatment for 10 min. The samples were collected, and the phosphorylation levels of IGF-1R, Akt and Erk were detected by Western Blot, and the dose-effect curve and time-effect curve of the phosphorylation levels of IGF-1R, Akt and Erk changed with CTS treatment were prepared.
[0031] Western Blot results such as Figure 4 As shown, it can be seen from it that PC12 cells were treated with different concentrations of CTS for 40 minutes, and as the concentration of CTS increased, the phosphorylation level of IGF-1R gradually decreased in a dose-dependent manner. The phosphorylation...
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