【摘要】：目的:研究牛磺酸對(duì)體外培養(yǎng)的乳腺癌細(xì)胞和裸鼠乳腺癌動(dòng)物模型的干預(yù)及機(jī)制。方法:體外實(shí)驗(yàn):A組對(duì)照組乳腺癌細(xì)胞用添加了10%胎牛血清和1%青鏈霉素雙抗的L-15細(xì)胞培養(yǎng)基培養(yǎng),B組乳腺癌細(xì)胞用添加了1%牛磺酸、10%胎牛血清和1%青鏈霉素雙抗的L-15細(xì)胞培養(yǎng)基培養(yǎng),繪制細(xì)胞周期生長(zhǎng)曲線并用流式細(xì)胞術(shù)測(cè)定細(xì)胞周期。體內(nèi)實(shí)驗(yàn):15只5周齡SPF級(jí)別雌性裸鼠隨機(jī)分成三組,1組接種乳腺癌細(xì)胞,給予正常飲水,2組接種乳腺癌細(xì)胞,給予添加了3%的�；撬犸嬎�,3組接種添加了1%牛磺酸的培養(yǎng)基培養(yǎng)的乳腺癌細(xì)胞,給予正常飲水,每隔2天用游標(biāo)卡尺測(cè)定記錄腫瘤體積,繪制腫瘤體積生長(zhǎng)曲線,40天后處理裸鼠,統(tǒng)計(jì)三組腫瘤質(zhì)量和體積,用蛋白免疫印跡方法測(cè)定乳腺癌瘤體中雌激素受體(estrogen receptor,ER)、孕激素受體(progesterone receptor,PR)、人類表皮生長(zhǎng)因子受體2(human epidemic growth factor receptor 2,HER2),鳥苷三磷酸酶激活蛋白(guanosine triphosphatase activating protein,p21ras)表達(dá)量的變化。結(jié)果:體外實(shí)驗(yàn)結(jié)果:細(xì)胞生長(zhǎng)曲線結(jié)果,b組乳腺癌細(xì)胞生長(zhǎng)速度比a組乳腺癌細(xì)胞生長(zhǎng)速度慢。流式細(xì)胞術(shù)檢測(cè)結(jié)果:b組g1期乳腺癌細(xì)胞比例為(89.93±1.68%),a組g1期細(xì)胞比例(84.9±2.08%)(p0.05),b組s期乳腺癌細(xì)胞比例為(66.54±2.17%)((p0.001)),a組s期細(xì)胞數(shù)比例(66.0±1.21%)(p0.001),b組g2期細(xì)胞比例為(.68±0.15%)(p0.001),a組的g2細(xì)胞比例(5.36±0.73%)(p0.001),b組的m期乳腺癌細(xì)胞比例為(19.49±0.93%)(p0.001),a組m期細(xì)胞比例(17.59±0.35%)(p0.001)。體內(nèi)實(shí)驗(yàn)結(jié)果:1組腫瘤平均質(zhì)量(3.46±0.95g),2組腫瘤平均質(zhì)量(2.02±0.75g)(p0.05),腫瘤抑制率41.6%,3組的腫瘤平均質(zhì)量(1.35±1.23g)(p0.05),腫瘤抑制率61.09%。1組腫瘤的體積為(3.06±0.004cm3)大于2組的腫瘤體積(1.79±0.006cm3)(p0.01),大于3組的腫瘤體積(1.18±0.009cm3)(p0.05)。蛋白免疫印跡實(shí)驗(yàn)結(jié)果:與1組腫瘤組織相比,2組er蛋白表達(dá)量提高了134%(p0.05),3組er蛋白表達(dá)量提高了60%(p0.05),2組pr蛋白的表達(dá)量提高了62.3%(p0.05),3組pr蛋白表達(dá)量提高(p0.05),2組her-2蛋白表達(dá)量下降(p0.05),3組her-2蛋白表達(dá)量下降了55.4%(p0.05),2組p21ras表達(dá)量下降(p0.01),3組p21ras蛋白表達(dá)量下降(p0.01),綜上所述,�；撬岣深A(yù)的兩組er和pr蛋白表達(dá)量上升,her2和p21ras的蛋白表達(dá)量下降。結(jié)論:1、牛磺酸對(duì)體外乳腺癌細(xì)胞的生長(zhǎng)有抑制作用,其主要通過抑制細(xì)胞分裂周期g1和g2期,即有絲分裂完成到期dna復(fù)制之前的間隙時(shí)間和dna復(fù)制完成到有絲分裂開始,提示�；撬峥赡芡ㄟ^延長(zhǎng)乳腺癌細(xì)胞的g1期和g2期來(lái)抑制細(xì)胞的分裂,進(jìn)而抑制乳腺癌細(xì)胞的生長(zhǎng)。2、牛磺酸通過促進(jìn)ER和PR蛋白的表達(dá)量,調(diào)節(jié)內(nèi)分泌環(huán)節(jié),降低HER2和p21ras蛋白表達(dá)量,控制腫瘤細(xì)胞的分裂、增殖環(huán)節(jié),最終達(dá)到牛磺酸抑制裸鼠乳腺癌腫瘤的生長(zhǎng)。
[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 鹵1.68%) in), a group (84.9 鹵2.08%) (p0.05). The percentage of breast cancer cells in the second phase of group b was (66.54 鹵2.17%) in) (p0.001), a group (66.0 鹵1.21%) and that in g2 phase in p0.001), b group was (.68 鹵0.15%) (p0.001). The percentage of g2 cells in group a was (5.36 鹵0.73%) (19.49 鹵0.93%) in), b group (17.59 鹵0.35%) in p0.001), a group. The results of in vivo experiments showed that the mean mass of tumor was (3.46 鹵0.95g) in group 1, (2.02 鹵0.75g) (p0.05) in group 2, and (1.35 鹵1.23g) (p0.05) in group 41.6. The tumor volume of group 61.09.1 was (3.06 鹵0.004cm3) larger than that of group 2 (1.79 鹵0.006cm3) (p0.01), and larger than that of group 3 (1.18 鹵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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.9

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