小分子化合物YM155抗食管癌作用机理及候选肿瘤标志物研究
发布时间：2019-02-15 17:49
【摘要】：食管癌是常见的恶性肿瘤之一,其早期症状不明显,诊断时多为中晚期,5年生存率低。晚期食管癌治疗方案基于放化疗,其中常用化疗药物为顺铂和5-氟尿嘧啶,其效果存在一定的局限性。相关研究表明,Survivin在肿瘤中高表达,靶向survivin的小分子抑制剂YM155具有显著抗肿瘤效应,然而YM155在食管癌中抗肿瘤的相关机理尚不清楚,有待深入研究。本论文由两部分组成,论文第一部分揭示靶向Survivin的小分子抑制剂YM155可有效抑制食管癌细胞的增殖,导致细胞周期停滞和活性氧生成增多,诱导食管癌细胞死亡,分子机制研究显示YM155诱导DNA损伤,触发PARP-1高度活化,形成poly-ADP聚合物和导致AIF从线粒体转位到细胞核,发生parthanatos的死亡。通过干扰PARP-1和AIF表达后降低了食管癌细胞对YM155敏感性,提示PARP-1和AIF在YM155诱导的食管癌细胞parthanatos死亡中发挥重要作用。转录组学研究显示,YM155处理可诱导食管癌细胞190个基因表达升高,230个基因表达下降,涉及细胞周期和细胞代谢等多个信号通路,提示YM155未诱导细胞凋亡相关基因改变。小鼠移植瘤揭示YM155可抑制小鼠体内移植瘤生长。论文第一部分发现并揭示了YM155诱导PARP活化介导细胞死亡的分子作用机理,提示YM155可作为一种治疗食管癌的潜在化疗药物。论文第二部分基于实验室前期关于食管癌与癌旁差异蛋白组学的研究中,筛选的候选差异蛋白Stathmin,进行了大量血清学的验证研究。首先,在74例食管癌患者和81例健康对照样本中验证,利用ELISA检测血清Stathmin表达,结果显示食管癌患者中血清Stathmin(6.9±0.44ng/mL)显著高于健康对照(0.73±0.06ng/mL,P0.0001),临床参数分析显示与食管癌淋巴结转移呈正相关(P=0.036)。其次,将血清Stathmin标志物与常用肿瘤标志物Cyfra21-1、CEA、CA72-4口CA19-9比较,血清Stathmin显示更高的灵敏度和特异度。最后,我们扩大食管癌样本至535例及288例健康对照,ESCC患者血清Stathmin(5.98±2.89ng/mL)仍显著高于健康对照(2.16±1.19 ng/mL, P0.0001),与淋巴结转移(P=0.029)和肿瘤大小均呈正相关(P=0.046)。提示血清Stathmin可作为辅助食管癌诊断的肿瘤标志物。
[Abstract]:Esophageal cancer is one of the common malignant tumors, its early symptoms are not obvious, diagnosis is mostly advanced, 5-year survival rate is low. The treatment of advanced esophageal cancer is based on radiotherapy and chemotherapy. The commonly used chemotherapy drugs are cisplatin and 5 fluorouracil. Related studies have shown that Survivin is highly expressed in tumors, and YM155, a small molecule inhibitor of targeted survivin, has a significant antitumor effect. However, the mechanism of YM155 antitumor in esophageal cancer is still unclear and needs to be further studied. This thesis consists of two parts. In the first part, it is revealed that YM155, a small molecular inhibitor targeting Survivin, can effectively inhibit the proliferation of esophageal cancer cells, lead to cell cycle arrest and increase the production of reactive oxygen species, and induce the death of esophageal cancer cells. The molecular mechanism showed that YM155 induced DNA damage, triggered the high activation of PARP-1, formed poly-ADP polymers and resulted in the translocation of AIF from mitochondria to the nucleus, resulting in the death of parthanatos. By interfering with the expression of PARP-1 and AIF, the sensitivity of esophageal cancer cells to YM155 was decreased, suggesting that PARP-1 and AIF play an important role in YM155 induced parthanatos death of esophageal cancer cells. Transcriptome studies showed that YM155 treatment could induce the increase of 190 genes expression and the decrease of 230 genes expression in esophageal carcinoma cells, involving several signal pathways, such as cell cycle and cell metabolism, suggesting that YM155 did not induce apoptosis related gene changes. Mouse transplanted tumor revealed that YM155 could inhibit the growth of transplanted tumor in mice. In the first part, the molecular mechanism of PARP activation mediated cell death induced by YM155 was discovered and revealed, suggesting that YM155 can be used as a potential chemotherapeutic agent for esophageal cancer. In the second part of this paper, a large number of serological validation studies were carried out based on the pre-laboratory studies on differential proteomics of esophageal cancer and paracancerous differentially expressed proteins (Stathmin,). First, the expression of serum Stathmin was detected by ELISA in 74 patients with esophageal cancer and 81 healthy controls. The results showed that serum Stathmin (6.9 卤0.44ng/mL) in patients with esophageal cancer was significantly higher than that in healthy controls (0.73 卤0.06ng / mL). The clinical parameters were positively correlated with lymph node metastasis of esophageal carcinoma (P0. 036). Secondly, serum Stathmin markers showed higher sensitivity and specificity than Cyfra21-1,CEA,CA72-4 mouth CA19-9, a common tumor marker. Finally, the serum Stathmin of ESCC patients (5.98 卤2.89ng/mL) was significantly higher than that of healthy controls (2.16 卤1.19 ng/mL, P 0.0001). It was positively correlated with lymph node metastasis (P0. 029) and tumor size (P0. 046). The results suggest that serum Stathmin can be used as a tumor marker in the diagnosis of esophageal carcinoma.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1
本文编号：2423580
[Abstract]:Esophageal cancer is one of the common malignant tumors, its early symptoms are not obvious, diagnosis is mostly advanced, 5-year survival rate is low. The treatment of advanced esophageal cancer is based on radiotherapy and chemotherapy. The commonly used chemotherapy drugs are cisplatin and 5 fluorouracil. Related studies have shown that Survivin is highly expressed in tumors, and YM155, a small molecule inhibitor of targeted survivin, has a significant antitumor effect. However, the mechanism of YM155 antitumor in esophageal cancer is still unclear and needs to be further studied. This thesis consists of two parts. In the first part, it is revealed that YM155, a small molecular inhibitor targeting Survivin, can effectively inhibit the proliferation of esophageal cancer cells, lead to cell cycle arrest and increase the production of reactive oxygen species, and induce the death of esophageal cancer cells. The molecular mechanism showed that YM155 induced DNA damage, triggered the high activation of PARP-1, formed poly-ADP polymers and resulted in the translocation of AIF from mitochondria to the nucleus, resulting in the death of parthanatos. By interfering with the expression of PARP-1 and AIF, the sensitivity of esophageal cancer cells to YM155 was decreased, suggesting that PARP-1 and AIF play an important role in YM155 induced parthanatos death of esophageal cancer cells. Transcriptome studies showed that YM155 treatment could induce the increase of 190 genes expression and the decrease of 230 genes expression in esophageal carcinoma cells, involving several signal pathways, such as cell cycle and cell metabolism, suggesting that YM155 did not induce apoptosis related gene changes. Mouse transplanted tumor revealed that YM155 could inhibit the growth of transplanted tumor in mice. In the first part, the molecular mechanism of PARP activation mediated cell death induced by YM155 was discovered and revealed, suggesting that YM155 can be used as a potential chemotherapeutic agent for esophageal cancer. In the second part of this paper, a large number of serological validation studies were carried out based on the pre-laboratory studies on differential proteomics of esophageal cancer and paracancerous differentially expressed proteins (Stathmin,). First, the expression of serum Stathmin was detected by ELISA in 74 patients with esophageal cancer and 81 healthy controls. The results showed that serum Stathmin (6.9 卤0.44ng/mL) in patients with esophageal cancer was significantly higher than that in healthy controls (0.73 卤0.06ng / mL). The clinical parameters were positively correlated with lymph node metastasis of esophageal carcinoma (P0. 036). Secondly, serum Stathmin markers showed higher sensitivity and specificity than Cyfra21-1,CEA,CA72-4 mouth CA19-9, a common tumor marker. Finally, the serum Stathmin of ESCC patients (5.98 卤2.89ng/mL) was significantly higher than that of healthy controls (2.16 卤1.19 ng/mL, P 0.0001). It was positively correlated with lymph node metastasis (P0. 029) and tumor size (P0. 046). The results suggest that serum Stathmin can be used as a tumor marker in the diagnosis of esophageal carcinoma.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1
【参考文献】
相关期刊论文 前2条
1 王峰;王留兴;何伟;朱利楠;赵培荣;樊青霞;;Stathmin基因在食管鳞癌中的表达及生物学意义[J];南方医科大学学报;2010年07期
2 ;Survivin antisense compound inhibits proliferation and promotes apoptosis in liver cancer cells[J];World Journal of Gastroenterology;2005年02期
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