Combination of epigallocatechin-3-gallate and cerubidin and use thereof
A technology of epigallocatechin and gallate, which is applied in the fields of medicine and genetic engineering, can solve the problems of no combination of EGCG and DNR, and achieve the effect of enhancing anti-tumor effect, reducing cardiotoxicity and enriching resources
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Embodiment 1
[0053] Example 1 Synergism of EGCG on DNR Inhibiting Tumor Cell Proliferation
[0054] The MTT method was used to detect the effect of EGCG on the effect of DNR on inhibiting the proliferation of tumor cells. The selected concentration of EGCG (20 μM), under this concentration, EGCG hardly affects the growth of selected tumor cells when used alone. After 0.4μM DNR acted on HepG2 for 48 hours, the number of cell survival was 49.3% of that of the control cells without drug addition. When used in combination with 10μM and 20μM EGCG, the number of cell survival decreased to 40% and 33.2%. Compared with DNR acting alone, EGCG combined with DNR increased the inhibition of cell proliferation by 16%, and the synergistic effect was significant, and the synergistic effect had an EGCG concentration gradient effect ( figure 1 ). In SMMC7721, EGCG showed a similar synergistic effect. After DNR combined with 20 μM EGCG, the cell survival was reduced from 65.9% to 45.3%, and the inhibitor...
Embodiment 2
[0055] Example 2 EGCG enhances the effect of DNR on G2 / M arrest of tumor cells
[0056] The effect of EGCG on DNR-induced cell cycle arrest in G2 / M phase was further detected. HepG2 and SMMC7721 cells were also selected. 24 hours after HepG2 was treated with 0.03 and 0.04 μM DNR, compared with the control cells without drug, the cells in G2 / M phase increased from 8.37% to 35.9% and 52.6%, respectively, and a significant G2 / M phase cycle arrest occurred , and there is a concentration-dependence of DNR, the higher the concentration, the more cells in G2 / M phase. 10μM EGCG had no significant effect on the cell cycle, when the same concentration of EGCG was combined with 0.03μM DNR, the number of cells in G2 / M phase increased from 35.9% to 41.28%; when EGCG was combined with 0.04μM DNR, the number of cells in G2 / M phase increased from 52.6% to 61.55%. EGCG can enhance the effect of DNR on cell G2 / M phase arrest.
[0057] The results of SMMC7721 were also similar. When EGCG was...
Embodiment 3
[0059] Example 3 Effect of EGCG on the anti-tumor activity of DNR in Hep3B expressing exogenous CBR1
[0060] First, western blot was used to detect the expression of CBR1 in the stable strain, and the cell lysate was detected by anti-CBR1 antibody and anti-Myc antibody. Hep3B-CBR1 had both endogenous CBR1 expression and exogenous CBR1-Myc expression. Hep3B-pcDNA3 .1 Expression of only endogenous CBR1. Use β-actin to level the loading amount ( image 3 ). The results proved that Hep3B-CBR1 had higher expression of CBR1.
[0061] Next, the effect of overexpression of CBR1 on cell drug resistance was detected. Cell survival was detected by MTT as an indicator of cell resistance to the drug. Drugs treated the control group Hep3B-pcDNA3.1 and Hep3B-CBR1 under the same conditions, that is, with 0.4μM DNR for 48 hours, compared with Hep3B-pcDNA3.1, the survival of Hep3B-CBR1 cells increased from 34.4% to 52.8% respectively , similarly, the survival of Hep3B-CBR1 cells increased...
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