Bcl-3在乳腺癌中的表达以及对癌细胞的增殖与凋亡的影响

发布时间：2018-02-01 14:38

本文关键词： Bcl-3 乳腺癌增殖凋亡　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨Bcl-3在乳腺癌中表达情况以及其下调后对乳癌细胞MDA-MB-231生物学习性的影响。方法:(1)选取80例乳腺癌组织以及对应癌旁组织,采用免疫组化法检测其中Bcl-3蛋白表达,并分析癌组织中Bcl-3的表达水平与80例乳腺癌患者临床病理参数之间的关系;(2)将乳腺癌MDA-MB-231细胞随机分为空白对照组(常规培养),阴性对照组(转染对照siRNA)和Bcl-3 siRNA组(转染Bcl-3 siRNA)。(3)转染48 h后,Western blotting法检测三组细胞Bcl-3的表达水平以鉴定转染效率;(4)利用MTT实验检测空白对照组、阴性对照组和Bcl-3 siRNA组三组细胞增殖能力;(5)利用平板克隆实验检测空白对照组、阴性对照组和Bcl-3 siRNA组三组细胞克隆形成能力;(6)利用流式细胞凋亡检测并分析空白对照组、阴性对照组和Bcl-3 siRNA组三组细胞凋亡率的差别。结果:(1)Bcl-3蛋白在乳腺癌以及对应癌旁组织中的阳性表达率分别为72.5%和51.3%,癌组织中阳性表达率明显高于对应的癌旁组织(P0.05);(2)80例乳腺癌组织中,Bcl-3蛋白的平均表达水平与该80例乳腺癌组织的组织学分级、淋巴结转移以及Her-2的表达水平呈正相关(P0.05),但是其表达水平与年龄大小、肿瘤直径大小、病理分型,以及雌、孕激素受体的表达水平无关(P0.05)。(3)Western blotting实验检测结果显示,在转染48 h后,空白对照组和阴性对照组两组乳腺癌MDA-MB-231细胞的Bcl-3基因的表达水平明显高于Bcl-3siRNA组的乳腺癌MDA-MB-231细胞内的表达(P均0.01),说明转染成功。(4)MTT法实验检测结果显示,与两对照组相比,Bcl-3 siRNA组细胞的增殖率明显降低(P0.01);(5)平板克隆实验结果显示,与两对照组相比,Bcl-3 siRNA组细胞的细胞克隆形成率明显降低(P0.01);(6)流式细胞凋亡实验结果显示,与两对照组相比,Bcl-3 siRNA组细胞的细胞凋亡率增高(P0.01)。结论:(1)Bcl-3在乳癌组织中呈高表达水平,而在对应癌旁组织中却呈低表达水平;(2)沉默Bcl-3后,MDA-MB-231乳腺癌细胞的增殖能力受到明显抑制。(3)沉默Bcl-3后,MDA-MB-231乳腺癌细胞的细胞克隆形成率受到显著的抑制作用。(4)沉默Bcl-3后,MDA-MB-231乳腺癌细胞的细胞凋亡率明显增高。
[Abstract]:Objective: To investigate the expression of Bcl-3 in breast cancer and its downregulation on MDA-MB-231 breast cancer cell biological study effect. Methods: (1) a total of 80 cases of breast cancer tissues and the corresponding adjacent tissues by immunohistochemical method to detect the expression of Bcl-3 protein, and to analyze the relationship between the expression of Bcl-3 in cancer tissue and in 80 cases of breast cancer patients with clinical pathological parameters; (2) MDA-MB-231 breast cancer cells were randomly divided into control group (routine culture), negative control group (cells transfected with siRNA) and Bcl-3 siRNA group (transfected with Bcl-3 siRNA). (3) 48 h after transfection, the expression level of detection of Western blotting method in three groups Bcl-3 cells to identify the transfection efficiency; (4) using the MTT assay, the blank control group, negative control group and Bcl-3 group siRNA three group cell proliferation; (5) using the assay colony blank control group, negative control group and Bcl-3 group siRNA three group The clone formation ability; (6) using flow cytometry apoptosis detection and analysis of the blank control group, negative control group and Bcl-3 group siRNA three group differences in the rate of apoptosis. Results: (1) the positive expression of Bcl-3 protein in breast carcinoma and corresponding adjacent tissues respectively 72.5% and 51.3%, carcinoma the positive expression rate was significantly higher than that in paracancerous tissues (P0.05); (2) in 80 cases of breast cancer, the average grade and the expression level of Bcl-3 protein in 80 cases of breast cancer tissue, lymph node metastasis and the expression of Her-2 was positive (P0.05), but its expression level with age. The diameter of the tumor size, pathological type, and estrogen, progesterone receptor expression is independent of the (P0.05) Western blotting (3). The results indicated that, at 48 h after transfection, the blank control group and negative control group, two groups of breast cancer MDA-MB-231 cells Bcl-3 gene expression The level of expression of MDA-MB-231 in breast cancer cells was significantly higher than that in group Bcl-3siRNA (P < 0.01), indicating the successful transfection. (4) the results of MTT test showed that, compared with two in the control group, Bcl-3 siRNA group significantly decreased the rate of cell proliferation (P0.01); (5) clone experimental results show that, compared with and two in the control group, Bcl-3 group siRNA cell clone cell formation was significantly reduced (P0.01); (6) apoptosis by flow cytometry experiment results show that, compared with two in the control group, apoptosis of Bcl-3 cells in siRNA group was increased (P0.01). Conclusion: (1) Bcl-3 in breast cancer tissues showed high expression level, but showed low expression level in the tumor tissues; (2) after silencing Bcl-3, MDA-MB-231 breast cancer cell proliferation was markedly inhibited. (3) after silencing Bcl-3, MDA-MB-231 cell clones of breast cancer cells by formation rate was significantly inhibited. (4) after silencing Bcl-3, MDA-MB-231 The apoptosis rate of breast cancer cells was significantly higher.

【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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