本文選題：乳腺癌 + PIB5PA��； 參考：《安徽醫(yī)科大學(xué)》2015年碩士論文

【摘要】：研究背景和目的：乳腺癌是女性常見的惡性腫瘤,其中有15%稱為三陰性乳腺癌(Triple-negative breast cancer,TNBC),即不表達(dá)雌激素受體、孕激素受體和人表皮生長因子受體2。目前國內(nèi)外對(duì)TNBC的研究進(jìn)展較為緩慢,化療仍然是其患者術(shù)后治療的主要手段。紫杉醇作為抗微管化療藥物,是TNBC術(shù)后的首選臨床化療藥物,然而臨床應(yīng)用中仍有很大部分TNBC對(duì)此耐藥,其機(jī)制可能與TNBC細(xì)胞內(nèi)PI3K/Akt信號(hào)通路激活有關(guān)。PIB5PA可抑制PI3K/Akt信號(hào)通路的活化,但其是否能通過此途徑來增強(qiáng)三陰性乳腺癌細(xì)胞對(duì)紫杉醇的敏感性尚無明確的答案,也是該課題的研究重點(diǎn)。方法：體外培養(yǎng)乳腺癌細(xì)胞株MCF-7和MDA-MB-231,采用MTT比色法觀察不同濃度(0,0.1,0.2,0.3,0.4,0.5μmol·L-1)紫杉醇處理后對(duì)兩種細(xì)胞株生存率的影響；流式細(xì)胞儀PI單染法檢測不同濃度紫杉醇處理MCF-7和MDA-MB-23148h后對(duì)細(xì)胞凋亡率的影響；應(yīng)用免疫蛋白印跡法(Western blot)檢測I0.3μmol·L-1紫杉醇處理MCF-7和MDA-MB-231不同時(shí)間點(diǎn)Bcl-2家族成員蛋白表達(dá)情況。將pCGN-PIB5PA質(zhì)粒轉(zhuǎn)染對(duì)紫杉醇相對(duì)不敏感人乳腺癌細(xì)胞MDA-MB-231,同時(shí)設(shè)立對(duì)照組,0.3μpmol·L-1紫杉醇處理24h后,免疫蛋白印跡法檢測PIB5PA,磷酸化Akt以及Bcl-2家族成員蛋白的表達(dá)情況,采用MTT法和PI單染法檢測PIB5PA轉(zhuǎn)染MDA-MB-231后對(duì)紫杉醇敏感性的影響。結(jié)果：選取兩株乳腺癌細(xì)胞株MCF-7和MDA-MB-231經(jīng)過紫杉醇一起培養(yǎng)后,發(fā)現(xiàn)紫杉醇能明顯誘導(dǎo)MCF-7細(xì)胞的凋亡,然而MDA-MB-231細(xì)胞株對(duì)紫杉醇相對(duì)不敏感；與此現(xiàn)象相對(duì)應(yīng)的是,紫杉醇誘導(dǎo)MCF-7細(xì)胞中Bim蛋白的明顯上調(diào),而MDA-MB-231細(xì)胞株經(jīng)紫杉醇處理后Bim表達(dá)幾乎沒有變化；過表達(dá)PIB5PA后三陰性細(xì)胞株MDA-MB-231對(duì)紫杉醇敏感性明顯增強(qiáng),表現(xiàn)為轉(zhuǎn)染后MDA-MB-231細(xì)胞的生存率明顯降低,同時(shí)其凋亡率明顯升高；同時(shí)伴隨著細(xì)胞中磷酸化Akt水平降低,Bim的表達(dá)增高。結(jié)論：PIB5PA能顯著增強(qiáng)紫杉醇誘導(dǎo)MDA-MB-231細(xì)胞的凋亡；PIB5PA在MDA-MB-231細(xì)胞凋亡中的作用可能與抑制磷酸化脂,上調(diào)Bim的表達(dá)有關(guān)。
[Abstract]:Background and objective: breast cancer is a common malignant tumor in women, 15% of which are called triple-negative breast cancer TNBC, that is, estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2. At present, the progress of TNBC research at home and abroad is slow, chemotherapy is still the main method of postoperative treatment. Paclitaxel, as an anti-microtubule chemotherapeutic drug, is the first choice of clinical chemotherapeutic drugs after TNBC surgery. The mechanism may be related to the activation of PI3K / Akt signaling pathway in TNBC cells. PIB5PA can inhibit the activation of PI3K / Akt signaling pathway, but there is no clear question whether PI3K / Akt signaling pathway can enhance the sensitivity of triple-negative breast cancer cells to paclitaxel. It is also the research focus of this subject. Methods: breast cancer cell lines MCF-7 and MDA-MB-231 were cultured in vitro. The survival rate of breast cancer cell line MCF-7 and MDA-MB-231treated with paclitaxel was observed by MTT colorimetry. The apoptotic rate of MCF-7 and MDA-MB-23148 h treated with different concentrations of paclitaxel was detected by flow cytometry Pi single staining, and the expression of Bcl-2 family members was detected by Western blotting assay (blot) at different time points treated with I0.3 渭 mol L-1 paclitaxel and MDA-MB-231 at different time points. PCGN-PIB5PA plasmid was transfected into human breast cancer cell line MDA-MB-231, which was relatively insensitive to paclitaxel, and the expression of PIB5PA, phosphorylated Akt and Bcl-2 family members were detected by Western blot after treated with paclitaxel at 0.3 渭 pmol L-1 for 24 hours. MTT assay and Pi single staining were used to detect the effect of PIB5PA transfection on the sensitivity of paclitaxel to MDA-MB-231. Results: two breast cancer cell lines MCF-7 and MDA-MB-231 were cultured with paclitaxel. It was found that paclitaxel could induce apoptosis of MCF-7 cells, but MDA-MB-231 cell line was relatively insensitive to paclitaxel. Bim protein was up-regulated in MCF-7 cells induced by paclitaxel, but the expression of Bim in MDA-MB-231 cells was almost unchanged after treatment with paclitaxel, and the sensitivity of MDA-MB-231 cells to paclitaxel was significantly increased after overexpression of PIB5PA. The results showed that the survival rate and apoptosis rate of MDA-MB-231 cells decreased significantly after transfection, and the level of phosphorylated Akt decreased with the increase of Bim expression in MDA-MB-231 cells. Conclusion the effect of PIB5PA on apoptosis of MDA-MB-231 cells induced by paclitaxel may be related to the inhibition of phosphorylated lipid and the up-regulation of Bim expression in MDA-MB-231 cells.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R737.9

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1 張楠楠;PIB5PA在增強(qiáng)三陰性乳腺癌細(xì)胞MDA-MB-231對(duì)紫杉醇的敏感性中的作用及其機(jī)制研究[D];安徽醫(yī)科大學(xué);2015年

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本文編號(hào)：2089864