小分子化合物YM155抗食管癌作用機(jī)理及候選腫瘤標(biāo)志物研究
發(fā)布時(shí)間：2019-02-15 17:49
【摘要】：食管癌是常見(jiàn)的惡性腫瘤之一,其早期癥狀不明顯,診斷時(shí)多為中晚期,5年生存率低。晚期食管癌治療方案基于放化療,其中常用化療藥物為順鉑和5-氟尿嘧啶,其效果存在一定的局限性。相關(guān)研究表明,Survivin在腫瘤中高表達(dá),靶向survivin的小分子抑制劑YM155具有顯著抗腫瘤效應(yīng),然而YM155在食管癌中抗腫瘤的相關(guān)機(jī)理尚不清楚,有待深入研究。本論文由兩部分組成,論文第一部分揭示靶向Survivin的小分子抑制劑YM155可有效抑制食管癌細(xì)胞的增殖,導(dǎo)致細(xì)胞周期停滯和活性氧生成增多,誘導(dǎo)食管癌細(xì)胞死亡,分子機(jī)制研究顯示YM155誘導(dǎo)DNA損傷,觸發(fā)PARP-1高度活化,形成poly-ADP聚合物和導(dǎo)致AIF從線粒體轉(zhuǎn)位到細(xì)胞核,發(fā)生parthanatos的死亡。通過(guò)干擾PARP-1和AIF表達(dá)后降低了食管癌細(xì)胞對(duì)YM155敏感性,提示PARP-1和AIF在YM155誘導(dǎo)的食管癌細(xì)胞parthanatos死亡中發(fā)揮重要作用。轉(zhuǎn)錄組學(xué)研究顯示,YM155處理可誘導(dǎo)食管癌細(xì)胞190個(gè)基因表達(dá)升高,230個(gè)基因表達(dá)下降,涉及細(xì)胞周期和細(xì)胞代謝等多個(gè)信號(hào)通路,提示YM155未誘導(dǎo)細(xì)胞凋亡相關(guān)基因改變。小鼠移植瘤揭示YM155可抑制小鼠體內(nèi)移植瘤生長(zhǎng)。論文第一部分發(fā)現(xiàn)并揭示了YM155誘導(dǎo)PARP活化介導(dǎo)細(xì)胞死亡的分子作用機(jī)理,提示YM155可作為一種治療食管癌的潛在化療藥物。論文第二部分基于實(shí)驗(yàn)室前期關(guān)于食管癌與癌旁差異蛋白組學(xué)的研究中,篩選的候選差異蛋白Stathmin,進(jìn)行了大量血清學(xué)的驗(yàn)證研究。首先,在74例食管癌患者和81例健康對(duì)照樣本中驗(yàn)證,利用ELISA檢測(cè)血清Stathmin表達(dá),結(jié)果顯示食管癌患者中血清Stathmin(6.9±0.44ng/mL)顯著高于健康對(duì)照(0.73±0.06ng/mL,P0.0001),臨床參數(shù)分析顯示與食管癌淋巴結(jié)轉(zhuǎn)移呈正相關(guān)(P=0.036)。其次,將血清Stathmin標(biāo)志物與常用腫瘤標(biāo)志物Cyfra21-1、CEA、CA72-4口CA19-9比較,血清Stathmin顯示更高的靈敏度和特異度。最后,我們擴(kuò)大食管癌樣本至535例及288例健康對(duì)照,ESCC患者血清Stathmin(5.98±2.89ng/mL)仍顯著高于健康對(duì)照(2.16±1.19 ng/mL, P0.0001),與淋巴結(jié)轉(zhuǎn)移(P=0.029)和腫瘤大小均呈正相關(guān)(P=0.046)。提示血清Stathmin可作為輔助食管癌診斷的腫瘤標(biāo)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 鹵0.44ng/mL) in patients with esophageal cancer was significantly higher than that in healthy controls (0.73 鹵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 鹵2.89ng/mL) was significantly higher than that of healthy controls (2.16 鹵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.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.1
本文編號(hào)：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 鹵0.44ng/mL) in patients with esophageal cancer was significantly higher than that in healthy controls (0.73 鹵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 鹵2.89ng/mL) was significantly higher than that of healthy controls (2.16 鹵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.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.1
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 王峰;王留興;何偉;朱利楠;趙培榮;樊青霞;;Stathmin基因在食管鱗癌中的表達(dá)及生物學(xué)意義[J];南方醫(yī)科大學(xué)學(xué)報(bào);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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