盐酸小檗碱对人乳腺癌MCF-7细胞裸鼠移植瘤的作用研究

发布时间：2018-05-17 22:09

本文选题：小檗碱 + 乳腺癌　；参考：《南方医科大学》2017年硕士论文

【摘要】：背景乳腺癌是一种女性常见的恶性肿瘤,严重影响身心健康甚至危害生命,其在女性癌症发病率中位居第一。乳腺癌的治疗主要以手术为主,并且配合术后化疗、靶向治疗等综合疗法,但放、化疗过程中的不良反应以及多药物耐药现象所导致的治疗失败仍是临床需进一步解决的问题。因此,寻找有效、低毒、天然、副作用小的中药治疗乳腺癌及其相关的研究已成为一个紧迫的问题。小檗碱又名黄连素,是一种常见的异喹啉类生物碱,从黄莲、黄柏等中草药中提取获得,具有抗病原微生物和抗菌的功效。小檗碱可以通过促进肿瘤细胞凋亡、抑制肿瘤细胞增殖、抑制肿瘤细胞转移以及增强肿瘤细胞对化疗药物的敏感性等方式起一定的抗肿瘤作用。小檗碱与乳腺癌之间的关系是目前的研究热点,体内研究尚未见报道。因此,本课题希望通过将人乳腺癌细胞MCF-7接种于裸鼠皮下,建立裸鼠乳腺癌动物模型,研究盐酸小檗碱对MCF-7乳腺癌小鼠体内产生的作用与影响。目的本课题通过将人乳腺癌细胞系MCF-7接种于裸鼠背部皮下,建立乳腺癌动物模型,研究盐酸小檗碱对MCF-7乳腺癌小鼠体内产生的作用与影响。方法一、在小鼠右背部皮下接种细胞,建立人乳腺癌MCF-7移植瘤裸鼠模型,计算出药物的抑瘤疗效(TGI)。二、应用HE染色法检测各实验组的裸鼠移植瘤组织的染色情况。三、应用TUNEL染色法检测各实验组的裸鼠移植瘤组织的细胞凋亡情况。四、应用流式细胞仪检测各实验组的裸鼠移植瘤组织的细胞周期情况。五、应用免疫组化SP法、Western blot检测各实验组的裸鼠移植瘤组织的MMP-2、MMP-9、p53、EGFR蛋白的表达情况。结果1、裸鼠移植瘤生长情况:(1)每组大鼠在第21天时的体重均比第1天有所下降,各组具有统计学差异(P0.05)。(2)五组裸鼠的体积差异有统计学意义(P0.05)。2、各药物组的肿瘤抑制率:B1、B2、B3、C组的TGI值分别是5.55%、4.91%、58.88%、67.72%,差异均有显著性差异(P0.01)。3、裸鼠移植瘤组织病理学变化:(1)肉眼:移植瘤呈圆形、类圆形或分叶状生长。(2)光镜:肿瘤细胞略增大,治疗组异型细胞减少,瘤组织结构破坏。4、TUNEL染色结果显示:各组间细胞凋亡指数比较有统计学差异(p0.01)。5、流式细胞术结果显示:(1)S期所占细胞周期分布经方差分析显示:各组间比较均有显著性差异(P0.01);(2)Go/G1期所占细胞周期分布经方差分析显示:各组间比较均有显著性差异(P0.01)。6、免疫组化、Western blot结果显示:MMP-2、MMP-9、P53、EGFR蛋白在对照组中的表达量显著高于其他实验组,其组间差异有统计学意义(P0.05)。结论1.成功建立了裸鼠人乳腺癌MCF-7细胞皮下移植瘤疾病模型。2.小檗碱能够抑制MCF-7乳腺癌裸鼠肿瘤生长,抑制作用呈浓度依赖性。
[Abstract]:Background breast cancer is a common malignant tumor in women, which seriously affects physical and mental health and even endangers life. It ranks first in the incidence of female cancer. The treatment of breast cancer is mainly based on surgery, combined with postoperative chemotherapy, targeted therapy and other comprehensive therapies, but the adverse reactions in the process of chemotherapy and the phenomenon of multi drug resistance are in the process of chemotherapy. Therefore, it has become an urgent problem to find effective, low toxic, natural, and minor side effects in the treatment of breast cancer and its related research. Berberine, also known as berberine, is a common isoquinoline, extracted from Chinese herbs such as yellow lotus and cortex Phellodendron. Berberine can inhibit tumor cell apoptosis, inhibit tumor cell proliferation, inhibit tumor cell metastasis and enhance the sensitivity of tumor cells to chemotherapeutic drugs. The relationship between berberine and breast cancer is a hot spot of research, and the study in vivo has not yet been studied Therefore, we hope to establish the animal model of breast cancer in nude mice by inoculating human breast cancer cell MCF-7 in nude mice and studying the effect and influence of berberine hydrochloride on MCF-7 breast cancer mice. Objective to establish the animal model of breast cancer by subcutaneous the human breast cancer cell line MCF-7 to the back of nude mice. Type, study the effect and influence of berberine hydrochloride on MCF-7 breast cancer mice. Method 1, inoculating cells in the right back of the mouse, establishing a nude mouse model of MCF-7 transplanted tumor of human breast cancer, and calculating the antitumor effect (TGI) of the drug (TGI). Two, HE staining method was used to detect the staining of the transplanted tumor tissue in the test groups. Three, TUN EL staining was used to detect the cell apoptosis in the transplanted tumor tissues of the experimental groups. Four, the cell cycle of the transplanted tumor tissues of the experimental groups was detected by flow cytometry. Five, the immunohistochemical SP method was used, and the expression of MMP-2, MMP-9, p53 and EGFR protein in the transplanted tumor tissues of the experimental groups was detected by Western blot. The results were 1, The growth of transplanted tumor in nude mice: (1) the weight of the rats in each group decreased at twenty-first days, with statistical difference (P0.05). (2) the volume difference between the five groups of nude mice was statistically significant (P0.05).2, the tumor inhibition rate of each group was B1, B2, B3, and the TGI values of group C were 5.55%, 4.91%, 58.88%, 67.72%, and the differences were significantly poor. Histopathological changes of xenograft tumor in nude mice: (1) naked eye: (1) naked eye: the transplanted tumor was round, round or lobulated. (2) light microscopy: tumor cells slightly increased, treatment group ISO cells decreased, tumor tissue structure destroyed.4, TUNEL staining results showed that the apoptosis index of each group was statistically different (P0.01).5, flow cytometry results The results showed: (1) the cell cycle distribution in the S period showed that there were significant differences between each group (P0.01). (2) the cell cycle distribution in Go/G1 period showed significant difference (P0.01).6, immunohistochemistry, and Western blot results showed that the expression of MMP-2, MMP-9, P53, EGFR protein in the control group Significantly higher than other experimental groups, the difference between the groups was statistically significant (P0.05). Conclusion 1. successfully established the nude mouse breast cancer MCF-7 cell subcutaneous transplantation tumor disease model.2. berberine can inhibit the growth of MCF-7 breast cancer in nude mice, and the inhibitory effect is concentration dependent.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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