西达本胺抑制骨髓瘤细胞自噬并促进DDR相关的凋亡反应
发布时间：2019-06-07 11:35
【摘要】：研究背景及目的:表观遗传学是近年来遗传学的研究热点。组蛋白氨基酸残端的甲基化、乙酰化、磷酸化、泛素化修饰等可以通过改变组蛋白和DNA的相互亲和力从而导致基因转录发生改变。多项研究表明,肿瘤的发生发展与组蛋白乙酰化状态(由组蛋白乙酰转移酶HAT和组蛋白去乙酰化酶HDAC共同调控)关系密切。组蛋白去乙酰化酶抑制剂(HDACi)作为一种特异性靶向抗肿瘤药物越来越引起人们的关注,目前FDA已批准HDAC抑制剂伏立诺他(Vorinostat,SAHA)和罗咪酯肽(Romidepsin)用于治疗皮肤及外周T细胞淋巴瘤,批准帕比司他(Panobinostat)治疗多发性骨髓瘤。多发性骨髓瘤(multiple myeloma,MM)是一种浆细胞恶性肿瘤,临床上经过DNA损伤性化疗、蛋白酶体抑制剂、免疫调节剂(沙利度胺或来那度胺)及大剂量化疗联合自体干细胞移植等治疗后,大多数患者最终仍出现疾病复发或者进展,因此,寻找更有效的治疗手段一直是MM治疗领域的研究热点。西达本胺是我国第一个自主研发的组蛋白去乙酰化酶抑制剂(HDACi),可以抑制HDAC1、2、3和10亚型的活性。在晚期肿瘤患者的Ⅰ期临床研究中,西达本胺体现了良好的耐受性,目前已有研究证明,西达本胺对肺癌、结肠癌、乳腺癌、肝癌、前列腺癌及淋巴瘤有抑制或杀伤活性,我国正在开展针对皮肤T细胞淋巴瘤(PTCL)及外周T细胞淋巴瘤(CTCL)的Ⅱ/Ⅲ期临床试验,初步统计结果提示疗效十分显著。综上述,HDACi(如伏立诺他、罗咪酯肽、帕比司他)均有抗MM活性,但单药治疗显示出了剂量依赖性的副作用,使得HDACi治疗骨髓瘤收到了限制;同时西达本胺治疗难治复发型PTCL显示出了极好的疗效。因此,我们设想MM的乙酰化状态与MM细胞活性密切相关,目前有关西达本胺抗MM活性尚未见相关报道,故本课题选择人多发性骨髓瘤细胞系RPMI8226、H929为实验对象,观察西达本胺对人多发性骨髓瘤细胞的抑制作用并探索其可能的机制。方法:①MTT法研究组蛋白去乙酰化酶抑制剂西达本胺对RPMI8226及H929细胞增殖的影响;②流式细胞术检测西达本胺作用后细胞凋亡率的变化及细胞周期分布情况;③RT-PCR检测西达本胺对MM细胞SIRT1基因、自噬相关基因m RNA表达的影响;④Western blot法检测西达本胺处理MM细胞48h后周期相关蛋白p21、自噬相关蛋白、DNA损伤相关蛋白及凋亡相关蛋白表达情况;⑤采用ATM激酶特异性抑制剂KU-55933抑制ATM激酶活性,研究ATM在西达本胺调控MM细胞增殖中的作用。结果:①西达本胺显著抑制MM细胞的增殖;②西达本胺上调p21蛋白,使细胞周期阻滞于G0/G1期;③西达本胺处理后的MM细胞DNA损伤相关蛋白γH2AX、p ATM及凋亡相关蛋白Cleaved Caspase-3表达水平升高;④KU-55933抑制ATM激酶活性后,部分逆转由西达本胺诱导的MM细胞DNA损伤和凋亡反应;⑤西达本胺抑制SIRT1基因的转录及表达,上调组蛋白H4K16的乙酰化水平;⑥西达本胺下调自噬相关基因ATG7和LC3的转录和表达。结论:①达本胺诱导MM细胞凋亡部分涉及DNA损伤反应;②西达本胺通过上调组蛋白H4K16乙酰化抑制MM细胞的自噬反应。
[Abstract]:The background and purpose of the study are as follows: Epigenetics is a hot topic of genetics in recent years. Methylation, ethylation, phosphorylation, ubiquitination modification, etc. of histone amino acid residues can lead to a change in gene transcription by changing the mutual affinity of histone and DNA. A number of studies have shown that the development of the tumor is closely related to histone ethylation status (which is co-regulated by histone B-transferase HAT and histone de-ethylation enzyme HDAC). Histone deethanolase inhibitors (HDACi) are increasingly concerned as a specific targeted anti-tumor drug, and currently the FDA has approved the HDAC inhibitor of VORinostat, SAHA and Romidepsin for the treatment of skin and peripheral T-cell lymphomas, Panobinstat was approved for multiple myeloma. Multiple myeloma (MM) is a plasma cell malignancy, and is clinically treated with DNA damage chemotherapy, proteasome inhibitor, immunomodulator (thalidomide or lenalidomide) and large-dose chemotherapy combined with autologous stem cell transplantation. In most patients, there is still a recurrence or progression of the disease, and therefore finding more effective means of treatment has been a hot spot in the field of MM therapy. Sida Benamine is the first histone de-ethylation enzyme inhibitor (HDACi) developed by China's first self-developed histone, which can inhibit the activity of HDAC1,2,3 and 10 subtypes. In the first clinical study of late-stage tumor patients, Sida Benamine is well tolerated and has been shown to have been shown to have inhibitory or anti-killing activity on lung cancer, colon cancer, breast cancer, liver cancer, prostate cancer and lymphoma, Our country is carrying out the 鈪，
本文编号：2494762
[Abstract]:The background and purpose of the study are as follows: Epigenetics is a hot topic of genetics in recent years. Methylation, ethylation, phosphorylation, ubiquitination modification, etc. of histone amino acid residues can lead to a change in gene transcription by changing the mutual affinity of histone and DNA. A number of studies have shown that the development of the tumor is closely related to histone ethylation status (which is co-regulated by histone B-transferase HAT and histone de-ethylation enzyme HDAC). Histone deethanolase inhibitors (HDACi) are increasingly concerned as a specific targeted anti-tumor drug, and currently the FDA has approved the HDAC inhibitor of VORinostat, SAHA and Romidepsin for the treatment of skin and peripheral T-cell lymphomas, Panobinstat was approved for multiple myeloma. Multiple myeloma (MM) is a plasma cell malignancy, and is clinically treated with DNA damage chemotherapy, proteasome inhibitor, immunomodulator (thalidomide or lenalidomide) and large-dose chemotherapy combined with autologous stem cell transplantation. In most patients, there is still a recurrence or progression of the disease, and therefore finding more effective means of treatment has been a hot spot in the field of MM therapy. Sida Benamine is the first histone de-ethylation enzyme inhibitor (HDACi) developed by China's first self-developed histone, which can inhibit the activity of HDAC1,2,3 and 10 subtypes. In the first clinical study of late-stage tumor patients, Sida Benamine is well tolerated and has been shown to have been shown to have inhibitory or anti-killing activity on lung cancer, colon cancer, breast cancer, liver cancer, prostate cancer and lymphoma, Our country is carrying out the 鈪，
本文编号：2494762
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