牛磺酸对乳腺癌细胞及其诱导的裸鼠乳腺癌肿瘤的影响及机制研究
发布时间：2018-11-20 19:14
【摘要】：目的:研究牛磺酸对体外培养的乳腺癌细胞和裸鼠乳腺癌动物模型的干预及机制。方法:体外实验:A组对照组乳腺癌细胞用添加了10%胎牛血清和1%青链霉素双抗的L-15细胞培养基培养,B组乳腺癌细胞用添加了1%牛磺酸、10%胎牛血清和1%青链霉素双抗的L-15细胞培养基培养,绘制细胞周期生长曲线并用流式细胞术测定细胞周期。体内实验:15只5周龄SPF级别雌性裸鼠随机分成三组,1组接种乳腺癌细胞,给予正常饮水,2组接种乳腺癌细胞,给予添加了3%的牛磺酸饮水,3组接种添加了1%牛磺酸的培养基培养的乳腺癌细胞,给予正常饮水,每隔2天用游标卡尺测定记录肿瘤体积,绘制肿瘤体积生长曲线,40天后处理裸鼠,统计三组肿瘤质量和体积,用蛋白免疫印迹方法测定乳腺癌瘤体中雌激素受体(estrogen receptor,ER)、孕激素受体(progesterone receptor,PR)、人类表皮生长因子受体2(human epidemic growth factor receptor 2,HER2),鸟苷三磷酸酶激活蛋白(guanosine triphosphatase activating protein,p21ras)表达量的变化。结果:体外实验结果:细胞生长曲线结果,b组乳腺癌细胞生长速度比a组乳腺癌细胞生长速度慢。流式细胞术检测结果:b组g1期乳腺癌细胞比例为(89.93±1.68%),a组g1期细胞比例(84.9±2.08%)(p0.05),b组s期乳腺癌细胞比例为(66.54±2.17%)((p0.001)),a组s期细胞数比例(66.0±1.21%)(p0.001),b组g2期细胞比例为(.68±0.15%)(p0.001),a组的g2细胞比例(5.36±0.73%)(p0.001),b组的m期乳腺癌细胞比例为(19.49±0.93%)(p0.001),a组m期细胞比例(17.59±0.35%)(p0.001)。体内实验结果:1组肿瘤平均质量(3.46±0.95g),2组肿瘤平均质量(2.02±0.75g)(p0.05),肿瘤抑制率41.6%,3组的肿瘤平均质量(1.35±1.23g)(p0.05),肿瘤抑制率61.09%。1组肿瘤的体积为(3.06±0.004cm3)大于2组的肿瘤体积(1.79±0.006cm3)(p0.01),大于3组的肿瘤体积(1.18±0.009cm3)(p0.05)。蛋白免疫印迹实验结果:与1组肿瘤组织相比,2组er蛋白表达量提高了134%(p0.05),3组er蛋白表达量提高了60%(p0.05),2组pr蛋白的表达量提高了62.3%(p0.05),3组pr蛋白表达量提高(p0.05),2组her-2蛋白表达量下降(p0.05),3组her-2蛋白表达量下降了55.4%(p0.05),2组p21ras表达量下降(p0.01),3组p21ras蛋白表达量下降(p0.01),综上所述,牛磺酸干预的两组er和pr蛋白表达量上升,her2和p21ras的蛋白表达量下降。结论:1、牛磺酸对体外乳腺癌细胞的生长有抑制作用,其主要通过抑制细胞分裂周期g1和g2期,即有丝分裂完成到期dna复制之前的间隙时间和dna复制完成到有丝分裂开始,提示牛磺酸可能通过延长乳腺癌细胞的g1期和g2期来抑制细胞的分裂,进而抑制乳腺癌细胞的生长。2、牛磺酸通过促进ER和PR蛋白的表达量,调节内分泌环节,降低HER2和p21ras蛋白表达量,控制肿瘤细胞的分裂、增殖环节,最终达到牛磺酸抑制裸鼠乳腺癌肿瘤的生长。
[Abstract]:Aim: to study the intervention and mechanism of taurine on breast cancer cells cultured in vitro and nude animal models of breast cancer. Methods: in vitro, breast cancer cells in group A were cultured on L-15 cell culture medium supplemented with 10% fetal bovine serum and 1% streptomycin double antibody, while breast cancer cells in group B were cultured with 1% taurine. 10% fetal bovine serum and 1% L-15 cell culture medium with 1% streptomycin double antibody were cultured. Cell cycle growth curve was plotted and cell cycle was measured by flow cytometry. In vivo experiment: fifteen 5-week-old female nude mice with SPF grade were randomly divided into three groups: the first group was inoculated with breast cancer cells, and the other two groups were inoculated with breast cancer cells, and 3% taurine was added to the drinking water. Three groups of breast cancer cells were inoculated with 1% taurine medium and were given normal drinking water. The tumor volume was recorded by Vernier caliper every 2 days and the growth curve of tumor volume was drawn. The nude mice were treated 40 days later. The tumor mass and volume of the three groups were measured by Western blot. Estrogen receptor (estrogen receptor,ER), progesterone receptor (progesterone receptor,PR) and human epidermal growth factor receptor 2 (human epidemic growth factor receptor 2 (HER2) were determined by Western blot. Changes in the expression of guanosine triphosphatase activator protein (guanosine triphosphatase activating protein,p21ras). Results: the results of in vitro experiment showed that the growth rate of group b was slower than that of group a. The results of flow cytometry showed that the percentage of breast cancer cells in group b was (89.93 卤1.68%) in), a group (84.9 卤2.08%) (p0.05). The percentage of breast cancer cells in the second phase of group b was (66.54 卤2.17%) in) (p0.001), a group (66.0 卤1.21%) and that in g2 phase in p0.001), b group was (.68 卤0.15%) (p0.001). The percentage of g2 cells in group a was (5.36 卤0.73%) (19.49 卤0.93%) in), b group (17.59 卤0.35%) in p0.001), a group. The results of in vivo experiments showed that the mean mass of tumor was (3.46 卤0.95g) in group 1, (2.02 卤0.75g) (p0.05) in group 2, and (1.35 卤1.23g) (p0.05) in group 41.6. The tumor volume of group 61.09.1 was (3.06 卤0.004cm3) larger than that of group 2 (1.79 卤0.006cm3) (p0.01), and larger than that of group 3 (1.18 卤0.009cm3) (p0.05). The results of Western blot showed that the expression of er protein was increased by 13.4% (p0.05) and the expression of er protein was increased by 60% (p0.05) in the two groups compared with that in the first group. The expression of pr protein increased by 62.3% (p0.05), the expression of pr protein increased (p0.05), and the expression of her-2 protein decreased (p0.05) in both groups. The expression of her-2 protein decreased by 55.4% (p0.05), the expression of p21ras decreased by 55.4% (p0. 05), the expression of p21ras protein decreased (p0. 01) in group 2, and the expression of p21ras protein decreased in group 3 (p0. 01). In conclusion, the expression of er and pr protein increased in the two groups treated with taurine. The protein expression of her2 and p21ras decreased. Conclusion: 1. Taurine can inhibit the growth of breast cancer cells in vitro by inhibiting the cell cycle G1 and g2, that is, the gap time before mitotic completion of dna replication and the completion of dna replication to the beginning of mitosis. It is suggested that taurine may inhibit cell division by prolonging the g1 and g2 phases of breast cancer cells and thus inhibit the growth of breast cancer cells. Taurine may regulate endocrine links by promoting the expression of ER and PR proteins. The expression of HER2 and p21ras protein was decreased, the division and proliferation of tumor cells were controlled, and taurine was used to inhibit the growth of breast cancer in nude mice.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9
[Abstract]:Aim: to study the intervention and mechanism of taurine on breast cancer cells cultured in vitro and nude animal models of breast cancer. Methods: in vitro, breast cancer cells in group A were cultured on L-15 cell culture medium supplemented with 10% fetal bovine serum and 1% streptomycin double antibody, while breast cancer cells in group B were cultured with 1% taurine. 10% fetal bovine serum and 1% L-15 cell culture medium with 1% streptomycin double antibody were cultured. Cell cycle growth curve was plotted and cell cycle was measured by flow cytometry. In vivo experiment: fifteen 5-week-old female nude mice with SPF grade were randomly divided into three groups: the first group was inoculated with breast cancer cells, and the other two groups were inoculated with breast cancer cells, and 3% taurine was added to the drinking water. Three groups of breast cancer cells were inoculated with 1% taurine medium and were given normal drinking water. The tumor volume was recorded by Vernier caliper every 2 days and the growth curve of tumor volume was drawn. The nude mice were treated 40 days later. The tumor mass and volume of the three groups were measured by Western blot. Estrogen receptor (estrogen receptor,ER), progesterone receptor (progesterone receptor,PR) and human epidermal growth factor receptor 2 (human epidemic growth factor receptor 2 (HER2) were determined by Western blot. Changes in the expression of guanosine triphosphatase activator protein (guanosine triphosphatase activating protein,p21ras). Results: the results of in vitro experiment showed that the growth rate of group b was slower than that of group a. The results of flow cytometry showed that the percentage of breast cancer cells in group b was (89.93 卤1.68%) in), a group (84.9 卤2.08%) (p0.05). The percentage of breast cancer cells in the second phase of group b was (66.54 卤2.17%) in) (p0.001), a group (66.0 卤1.21%) and that in g2 phase in p0.001), b group was (.68 卤0.15%) (p0.001). The percentage of g2 cells in group a was (5.36 卤0.73%) (19.49 卤0.93%) in), b group (17.59 卤0.35%) in p0.001), a group. The results of in vivo experiments showed that the mean mass of tumor was (3.46 卤0.95g) in group 1, (2.02 卤0.75g) (p0.05) in group 2, and (1.35 卤1.23g) (p0.05) in group 41.6. The tumor volume of group 61.09.1 was (3.06 卤0.004cm3) larger than that of group 2 (1.79 卤0.006cm3) (p0.01), and larger than that of group 3 (1.18 卤0.009cm3) (p0.05). The results of Western blot showed that the expression of er protein was increased by 13.4% (p0.05) and the expression of er protein was increased by 60% (p0.05) in the two groups compared with that in the first group. The expression of pr protein increased by 62.3% (p0.05), the expression of pr protein increased (p0.05), and the expression of her-2 protein decreased (p0.05) in both groups. The expression of her-2 protein decreased by 55.4% (p0.05), the expression of p21ras decreased by 55.4% (p0. 05), the expression of p21ras protein decreased (p0. 01) in group 2, and the expression of p21ras protein decreased in group 3 (p0. 01). In conclusion, the expression of er and pr protein increased in the two groups treated with taurine. The protein expression of her2 and p21ras decreased. Conclusion: 1. Taurine can inhibit the growth of breast cancer cells in vitro by inhibiting the cell cycle G1 and g2, that is, the gap time before mitotic completion of dna replication and the completion of dna replication to the beginning of mitosis. It is suggested that taurine may inhibit cell division by prolonging the g1 and g2 phases of breast cancer cells and thus inhibit the growth of breast cancer cells. Taurine may regulate endocrine links by promoting the expression of ER and PR proteins. The expression of HER2 and p21ras protein was decreased, the division and proliferation of tumor cells were controlled, and taurine was used to inhibit the growth of breast cancer in nude mice.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9
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