發(fā)布時(shí)間：2018-02-01 14:38

  本文關(guān)鍵詞： Bcl-3 乳腺癌 增殖 凋亡　出處：《江蘇大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討B(tài)cl-3在乳腺癌中表達(dá)情況以及其下調(diào)后對乳癌細(xì)胞MDA-MB-231生物學(xué)習(xí)性的影響。方法:(1)選取80例乳腺癌組織以及對應(yīng)癌旁組織,采用免疫組化法檢測其中Bcl-3蛋白表達(dá),并分析癌組織中Bcl-3的表達(dá)水平與80例乳腺癌患者臨床病理參數(shù)之間的關(guān)系;(2)將乳腺癌MDA-MB-231細(xì)胞隨機(jī)分為空白對照組(常規(guī)培養(yǎng)),陰性對照組(轉(zhuǎn)染對照siRNA)和Bcl-3 siRNA組(轉(zhuǎn)染Bcl-3 siRNA)。(3)轉(zhuǎn)染48 h后,Western blotting法檢測三組細(xì)胞Bcl-3的表達(dá)水平以鑒定轉(zhuǎn)染效率;(4)利用MTT實(shí)驗(yàn)檢測空白對照組、陰性對照組和Bcl-3 siRNA組三組細(xì)胞增殖能力;(5)利用平板克隆實(shí)驗(yàn)檢測空白對照組、陰性對照組和Bcl-3 siRNA組三組細(xì)胞克隆形成能力;(6)利用流式細(xì)胞凋亡檢測并分析空白對照組、陰性對照組和Bcl-3 siRNA組三組細(xì)胞凋亡率的差別。結(jié)果:(1)Bcl-3蛋白在乳腺癌以及對應(yīng)癌旁組織中的陽性表達(dá)率分別為72.5%和51.3%,癌組織中陽性表達(dá)率明顯高于對應(yīng)的癌旁組織(P0.05);(2)80例乳腺癌組織中,Bcl-3蛋白的平均表達(dá)水平與該80例乳腺癌組織的組織學(xué)分級、淋巴結(jié)轉(zhuǎn)移以及Her-2的表達(dá)水平呈正相關(guān)(P0.05),但是其表達(dá)水平與年齡大小、腫瘤直徑大小、病理分型,以及雌、孕激素受體的表達(dá)水平無關(guān)(P0.05)。(3)Western blotting實(shí)驗(yàn)檢測結(jié)果顯示,在轉(zhuǎn)染48 h后,空白對照組和陰性對照組兩組乳腺癌MDA-MB-231細(xì)胞的Bcl-3基因的表達(dá)水平明顯高于Bcl-3siRNA組的乳腺癌MDA-MB-231細(xì)胞內(nèi)的表達(dá)(P均0.01),說明轉(zhuǎn)染成功。(4)MTT法實(shí)驗(yàn)檢測結(jié)果顯示,與兩對照組相比,Bcl-3 siRNA組細(xì)胞的增殖率明顯降低(P0.01);(5)平板克隆實(shí)驗(yàn)結(jié)果顯示,與兩對照組相比,Bcl-3 siRNA組細(xì)胞的細(xì)胞克隆形成率明顯降低(P0.01);(6)流式細(xì)胞凋亡實(shí)驗(yàn)結(jié)果顯示,與兩對照組相比,Bcl-3 siRNA組細(xì)胞的細(xì)胞凋亡率增高(P0.01)。結(jié)論:(1)Bcl-3在乳癌組織中呈高表達(dá)水平,而在對應(yīng)癌旁組織中卻呈低表達(dá)水平;(2)沉默Bcl-3后,MDA-MB-231乳腺癌細(xì)胞的增殖能力受到明顯抑制。(3)沉默Bcl-3后,MDA-MB-231乳腺癌細(xì)胞的細(xì)胞克隆形成率受到顯著的抑制作用。(4)沉默Bcl-3后,MDA-MB-231乳腺癌細(xì)胞的細(xì)胞凋亡率明顯增高。
[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.

【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.9

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本文編號：1482098