左旋紫草素通过抑制蛋白酶体活性抗肿瘤作用的体内外分析-anti - tumor effect of levoshikonin by inhibiting proteasome activity in vivo and in vitro.docxVIP

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左旋紫草素通过抑制蛋白酶体活性抗肿瘤作用的体内外分析-anti - tumor effect of levoshikonin by inhibiting proteasome activity in vivo and in vitro.docx

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左旋紫草素通过抑制蛋白酶体活性抗肿瘤作用的体内外分析-anti - tumor effect of levoshikonin by inhibiting proteasome activity in vivo and in vitro

广州医学院硕士学位论文左旋紫草素通过抑制蛋白酶体活性抗肿瘤作用的体内外研究广州医学院硕士学位论文左旋紫草素通过抑制蛋白酶体活性抗肿瘤作用的体内外研究 PAGE PAGE 6 P388 cells were treated with various concentrations of Shikonin (0-6μ mol/L) for 6 hours respectively or treateded with the same dose of Shikonin (2μ mol/L) for different time points (0-12h). Then the cell lysates were incubated with Chymotrypsin- like peptide substrates. It was found that shikonin can inhibit proteasome activity in P388 cells in a dose and time dependent manner. In the same condition ,the death rates were detected by flow cytometry (Annexin V-FITC staining). It was found that P388 cells could be induced to death by Shikonin also in a dose and time dependent manner. Additionally, western blot assay found that shikonin could induce ubiquitination protein aggregation and PARP cutting in a concentration and time dependent manner.Shikonin induced proteasome inhibition occurs prior to tumor cell death. Shikonin’s effect on cumulative survival in KM mice bearing murine leukemia Male KM mice were injected with murine leukemia P388 cells.After 24 hour,mice were randomly divided into 2 groups and treated with i.p.bolus injections of either drug vehicle (V) or 4 mg/kg/day shikonin (Shi 4) for 7 consecutive days.The mice were then kept for additional 60 days to determine the effect on mice survival of shikonin. It was found 4 mg/kg shikonin treatment significantly extended the survival period of KM mice bearing murine leukemia.Within 23 days,the entire control vehicle treated mice died.In a sharp contrast,the 9 of 10 mice treated with 4 mg/kg shikonin survived by the end of the experiment (60 days) only one mice died in 28 days. Shikonin suppressed solid tumor growth and induced proteasome inhibition in vivo Murine hepatoma H22 cells were injected s.c.in the left armpit of each Male KM mice.After 24 hour of injection,mice were randomly divided into 3 groups and treated with either vehicle or shikonin (4.0 or 8.0 mg/kg) for 7 consecutive days.Two days later, the mice wer