大蒜素对小鼠黑色素瘤B16F10细胞体外增殖和迁移能力的影响

Effects of Allicin on Proliferation and Migration of Mouse Melanoma B16F10 Cells in vitro

旨在研究不同浓度的大蒜素对色素形成关键基因小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)表达的影响,以及对B16F10细胞增殖和迁移的作用,为家养宠物及畜牧行业中的动物黑色素瘤治疗提供一个新思路。将添加不同浓度大蒜素的黑色素瘤B16F10细胞分为对照组(0μg·mL^(-1))和试验组(5、10、15、20μg·mL^(-1))。采用qRT-PCR检测B16F10细胞中MITF mRNA的表达水平;Western blot检测MITF蛋白质的表达水平。通过吸光度值计算细胞的增殖率,进而确定不同浓度大蒜素对细胞增殖的抑制情况。不同浓度大蒜素处理B16F10细胞42 h后,采用细胞划痕评价细胞迁移能力。结果显示,随大蒜素浓度升高,B16F10细胞中色素形成关键基因MITF mRNA的表达量逐渐降低;Western blot结果显示,MITF蛋白表达量呈浓度依赖性降低。在细胞增殖检测中,发现5、10、15、20μg·mL^(-1)大蒜素对B16F10细胞的增殖能力具有显著的抑制效果(P<0.001),且抑制率随浓度升高而加强。细胞划痕结果显示,大蒜素浓度在10、15、20μg·mL^(-1)时对细胞的迁移抑制较大。综上表明,大蒜素能够影响黑色素瘤B16F10细胞特异性标记基因MITF的表达,并抑制了黑色素瘤B16F10细胞的增殖和迁移。

The study aimed to investigate the effects of different concentrations of allicin on the expression of microphthalmia-associated transcription factor(MITF)and its effects on the proliferation and migration of B16F10 cells,which provide a new idea for the treatment of animal melanoma in pet and animal husbandry.B16F10 cells were divided into control group(0μg·mL^(-1))and experimental group(5,10,15,20μg·mL^(-1))supplemented with different concentrations of allicin.The expression of MITF mRNA in B16F10 cells was detected by qRT-PCR and the expression of MITF protein was detected by Western blot.The cell growth rate was calculated by using the absorbance of cells at 515 nm,and then the inhibition rate of different concentrations of allicin on cell proliferation was determined.Wound healing was used to evaluate the effects of different concentrations of allicin on the migration of B16F10 cells after 42 hours.The results showed that the expression of MITF mRNA decreased gradually with the increase of allicin concentration,while Western blot results showed that the expression of MITF protein decreased in a concentration-dependent manner.Allicin at the concentrations of 5,10,15,20μg·mL^(-1) could significantly inhibit the proliferation of B16F10 cells(P<0.001),and the inhibition rate increased with the increase of allicin concentration.Wound healing results showed that allicin significantly inhibited the cell migration at the concentration of 10μg·mL^(-1),15μg·mL^(-1) and 20μg·mL^(-1).This experiment showed that allicin could affect the expression of the melanoma-specific marker gene MITF and inhibit the proliferation and migration of B16F10 cells.