本文選題：辛二酰苯胺異羥肟酸　切入點：組蛋白去乙�；敢种苿�　出處：《河北北方學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：卵巢癌是致死率最高的婦科惡性腫瘤，手術(shù)和化療是其主要治療手段。紫杉醇聯(lián)合鉑類是目前卵巢癌化療的基礎(chǔ)方案，雖然在一定程度上延長了患者生存期，但由于存在原發(fā)和繼發(fā)耐藥現(xiàn)象，復(fù)發(fā)率仍高達(dá)60%～70%，是導(dǎo)致卵巢癌治療失敗的重要原因。因此，尋找卵巢癌化療耐藥后的敏感新藥和優(yōu)化化療聯(lián)合方案勢在必行。 辛二酰苯胺異羥肟酸（Suberoylanilide hydroxamic acid, SAHA）是一種異羥肟酸類組蛋白去乙酰化酶抑制劑，它可通過抑制組蛋白去乙酰化酶活性，阻斷由組蛋白去乙酰化酶募集功能紊亂而導(dǎo)致的基因表達(dá)異常，發(fā)揮抗腫瘤作用。此外，研究證實，SAHA與多種化療藥物，如順鉑、三苯氧胺及硼替佐米聯(lián)合使用，發(fā)揮良好的協(xié)同效應(yīng)。目前尚沒有關(guān)于SAHA單獨及聯(lián)合PTX作用卵巢癌紫杉醇耐藥細(xì)胞株系統(tǒng)研究及機(jī)制探討。 本研究通過體外培養(yǎng)卵巢癌紫杉醇耐藥細(xì)胞株OC3/P，經(jīng)過MTT法檢測紫杉醇卵巢癌耐藥細(xì)胞對SAHA是否存在交叉耐藥，采用倒置顯微鏡、CCK-8、AnnexinV-FITC/PI雙染法分別檢測OC3/P細(xì)胞經(jīng)SAHA、PTX單獨及聯(lián)合處理后，細(xì)胞形態(tài)、存活率、凋亡的變化。通過Q-RT PCR進(jìn)一步檢測細(xì)胞內(nèi)凋亡相關(guān)基因bcl-2，bax及耐藥相關(guān)基因mdr1的表達(dá)變化，通過Western blot檢測凋亡蛋白caspase-3的表達(dá)。 結(jié)果表明：卵巢癌紫杉醇耐藥細(xì)胞株OC3/P對SAHA不存在交叉耐藥(P0.05)。SAHA聯(lián)合PTX較各自單獨應(yīng)用對細(xì)胞形態(tài)的影響及生長抑制作用顯著增強(qiáng)（P0.05）。此外，聯(lián)合組較藥物單用組細(xì)胞凋亡率顯著增高。Q-RT PCR結(jié)果顯示，，SAHA聯(lián)合PTX較各自單獨應(yīng)用可明顯減弱細(xì)胞內(nèi)bcl-2表達(dá)，同時增強(qiáng)凋亡基因bax表達(dá)(P0.05)。此外，細(xì)胞經(jīng)SAHA處理后，mdr1基因水平較對照組顯著下降(P0.05)。Western blot結(jié)果表明，SAHA單獨及聯(lián)合PTX作用細(xì)胞后，均可以增強(qiáng)細(xì)胞內(nèi)caspase-3蛋白表達(dá)。 結(jié)論提示： SAHA聯(lián)合PTX可有效抑制卵巢癌紫杉醇耐藥細(xì)胞OC3/P的存活并誘導(dǎo)其凋亡，兩藥物聯(lián)合有協(xié)同效應(yīng)。
[Abstract]:Ovarian cancer is the most lethal gynecologic malignant tumor, surgery and chemotherapy is the main treatment. Paclitaxel combined with platinum based chemotherapy for ovarian cancer program at present, although prolong the survival time of the patients to a certain extent, but due to the presence of primary and secondary resistance phenomenon, the recurrence rate is still as high as 60% to 70% that is an important cause of failure in the treatment of ovarian cancer. Therefore, it is imperative to find new drug sensitive and optimal chemotherapy regimens of ovarian cancer after chemotherapy.
Xin two anilide hydroxamic acid (Suberoylanilide hydroxamic, acid, SAHA) is a kind of hydroxamic acid histone deacetylase inhibitors, it can inhibit the activity of histone deacetylase, blocked by histone deacetylase recruitment disorders and gene expression led to abnormal, antitumor effect. In addition, studies confirmed that SAHA with a variety of chemotherapy drugs, such as cisplatin, tamoxifen and bortezomib in combination, play a good synergistic effect. There is no study on paclitaxel resistance alone and combined with PTX SAHA ovarian cancer cell line system and mechanism research.
This study by in vitro paclitaxel resistance of ovarian cancer cell line OC3/P, after MTT assay of paclitaxel resistant ovarian cancer cells are cross resistant to SAHA by inverted microscope, CCK-8, OC3/P were detected by SAHA AnnexinV-FITC/PI double staining cells, PTX alone and combined treatment, cell morphology, viability, apoptosis the changes of apoptosis related gene bcl-2. Further detection by Q-RT PCR, expression of Bax and multidrug resistance related gene MDR1, Western expression by blot detection of apoptosis protein caspase-3.
緇撴灉琛ㄦ槑錛氬嵉宸㈢檶绱潐閱囪

本文編號：1625941