辛二酰苯胺异羟肟酸联合紫杉醇对卵巢癌紫杉醇耐药细胞株的体外研究

发布时间：2018-03-17 18:28

  本文选题：辛二酰苯胺异羟肟酸　切入点：组蛋白去乙酰化酶抑制剂　出处：《河北北方学院》2014年硕士论文　论文类型：学位论文

【摘要】：卵巢癌是致死率最高的妇科恶性肿瘤，手术和化疗是其主要治疗手段。紫杉醇联合铂类是目前卵巢癌化疗的基础方案，虽然在一定程度上延长了患者生存期，但由于存在原发和继发耐药现象，复发率仍高达60%～70%，是导致卵巢癌治疗失败的重要原因。因此，寻找卵巢癌化疗耐药后的敏感新药和优化化疗联合方案势在必行。 辛二酰苯胺异羟肟酸（Suberoylanilide hydroxamic acid, SAHA）是一种异羟肟酸类组蛋白去乙酰化酶抑制剂，它可通过抑制组蛋白去乙酰化酶活性，阻断由组蛋白去乙酰化酶募集功能紊乱而导致的基因表达异常，发挥抗肿瘤作用。此外，研究证实，SAHA与多种化疗药物，如顺铂、三苯氧胺及硼替佐米联合使用，发挥良好的协同效应。目前尚没有关于SAHA单独及联合PTX作用卵巢癌紫杉醇耐药细胞株系统研究及机制探讨。 本研究通过体外培养卵巢癌紫杉醇耐药细胞株OC3/P，经过MTT法检测紫杉醇卵巢癌耐药细胞对SAHA是否存在交叉耐药，采用倒置显微镜、CCK-8、AnnexinV-FITC/PI双染法分别检测OC3/P细胞经SAHA、PTX单独及联合处理后，细胞形态、存活率、凋亡的变化。通过Q-RT PCR进一步检测细胞内凋亡相关基因bcl-2，bax及耐药相关基因mdr1的表达变化，通过Western blot检测凋亡蛋白caspase-3的表达。 结果表明：卵巢癌紫杉醇耐药细胞株OC3/P对SAHA不存在交叉耐药(P0.05)。SAHA联合PTX较各自单独应用对细胞形态的影响及生长抑制作用显著增强（P0.05）。此外，联合组较药物单用组细胞凋亡率显著增高。Q-RT PCR结果显示，，SAHA联合PTX较各自单独应用可明显减弱细胞内bcl-2表达，同时增强凋亡基因bax表达(P0.05)。此外，细胞经SAHA处理后，mdr1基因水平较对照组显著下降(P0.05)。Western blot结果表明，SAHA单独及联合PTX作用细胞后，均可以增强细胞内caspase-3蛋白表达。 结论提示： SAHA联合PTX可有效抑制卵巢癌紫杉醇耐药细胞OC3/P的存活并诱导其凋亡，两药物联合有协同效应。
[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.
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