Application of dimethyldiguanide in preparation of medicaments for preventing or treating hepatocellular carcinoma
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Embodiment 1
[0025] Example 1. The inhibitory effect of metformin on the growth of human hepatocellular carcinoma cell lines in vitro.
[0026] Hepatocellular carcinoma cell lines SMMC7721, Huh7, HCC-LM3, and HepG2 (purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences) in the logarithmic growth phase were seeded in 96-well plates at a density of 2000-3000 cells / well, and replaced with Complete medium containing metformin at different concentrations (with no metformin group as the control, four concentrations of metformin were 1mM, 2mM, 5mM, and 10mM), respectively, at different time points (48 hours and 96 hours) with Cell Counting Kit-8 (CCK-8 kit, purchased from Dojindo, Japan, to detect cell proliferation, for the detection method, see "Experimental Technology of Medical Immunology" (Soochow University Press, date of publication: February 2011).
[0027] The result is as figure 1 As shown, metformin has obvious inhibitory effect on the proliferation of different cel...
Embodiment 2
[0028] Example 2. The inhibitory effect of metformin on the colony formation ability of liver cancer cell lines.
[0029] Hepatocellular carcinoma cell lines SMMC7721, HCC-LM3, and HepG2 in the logarithmic growth phase were seeded in a 6-well plate at a density of 1000 cells / well, and replaced with metformin (set at two concentrations of 2mM and 5mM) after 24 hours. Complete culture medium, and the empty cell group without any stimulation as the control. After culturing in the cell incubator for 10 days, the formed cell clones were stained with crystal violet and counted, and the ratio of the number of cell clones formed by the metformin treatment group to the control group was calculated. The result is as Figure 4 As shown, the number of cell clones formed by the three types of cells was significantly reduced after metformin treatment, suggesting that metformin can inhibit the malignant transformation ability of liver cancer cells.
Embodiment 3
[0030] Example 3. The inhibitory effect of metformin combined with chemotherapy drugs on the growth of human hepatocellular carcinoma cell lines in vitro.
[0031] Hepatocellular carcinoma cell lines SMMC7721, HCC-LM3, and HepG2 in the logarithmic growth phase were seeded in 96-well plates at a density of 2000-3000 cells / well, and replaced with metformin and cisplatin (purchased from Sigma) after 24 hours. ) complete culture medium, set up a blank control (empty cells without any stimulation), a single metformin (2mM-5mM) control, and a single cisplatin (0.5-2ug / ml) control. After acting for 24 hours, the detection results by CCK-8 method are as follows: Figure 5 , when metformin and cisplatin were used in combination, the survival rate of liver cancer cell lines was significantly lower than that of cisplatin alone, suggesting that the combination of metformin and cisplatin can enhance the inhibitory effect on the growth of liver cancer cell lines.
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