美登木素抑制鼻咽癌細胞上皮間充質(zhì)轉(zhuǎn)化及其機理研究
發(fā)布時間：2019-06-03 05:32
【摘要】：鼻咽癌(nasopharyngeal carcinoma,NPC)是高發(fā)于中國南方尤其廣東地區(qū)的惡性腫瘤。針對早期和局部晚期鼻咽癌患者,目前主要的治療方式是化療或者放化療,這類患者的5年生存率可達到85%。但是,仍然有部分患者由于發(fā)生遠處轉(zhuǎn)移和局部復發(fā)而導致治療失敗。含鉑的雙藥化療是治療局部復發(fā)和遠處轉(zhuǎn)移鼻咽癌患者的標準治療方案。但是該方案存在緩解持續(xù)時間短,并發(fā)癥較多等局限。因此尋找新的治療方法和藥物以減少并發(fā)癥并提高局部復發(fā)和遠處轉(zhuǎn)移鼻咽癌患者的生存期仍然是亟待解決的臨床問題。隨著抗體-藥物偶聯(lián)物(Antibody-drug conjugate,ADC)技術(shù)的完善,一批抗腫瘤作用強,但具有一定毒性的藥物又重新得到了關(guān)注。美登木素是抗體-藥物偶聯(lián)物中應用最廣泛的細胞毒性藥物。在本研究中,我們將探討美登木素對鼻咽癌生物學功能的影響。首先我們通過生物信息學分析了美登木素相關(guān)的基因芯片,該芯片包含分別經(jīng)T-DM1(trastuzumab-emtansine,曲妥珠單抗-美登木素偶聯(lián)藥物)以及trastuzumab曲妥珠單抗處理的乳腺癌組織。通過比對兩組不同的芯片,我們可以得出美登木素衍生物DM1會影響乳腺癌細胞哪些基因的表達。最終我們發(fā)現(xiàn)DM1可以顯著抑制乳腺癌細胞中EMT相關(guān)基因的表達。相關(guān)研究亦表明,美登木素可以抑制乳腺癌細胞株EMT的發(fā)生�；诖�,我們進一步探索美登木素是否抑制了鼻咽癌細胞EMT的發(fā)生。通過MTT、平板克隆形成實驗、周期實驗以及相關(guān)周期蛋白的檢測、裸鼠皮下成瘤,我們初步證實了美登木素顯著抑制鼻咽癌細胞體內(nèi)外的增殖能力。隨后,通過劃痕實驗、Transwell以及Boyden,我們證實美登木素可以顯著抑制鼻咽癌細胞株侵襲遷移能力。因此,EMT的表型受到了美登木素的顯著影響。由此,進一步通過Western blot檢測EMT相關(guān)蛋白的變化,發(fā)現(xiàn)上皮性質(zhì)相關(guān)的標記物E-cadherin顯著升高,而間充質(zhì)細胞性質(zhì)相關(guān)的標記物N-cadherin以及Vimentin標記物降低。由此說明鼻咽癌細胞在美登木素作用下,EMT受到抑制并降低了細胞的侵襲遷移能力。腫瘤干細胞被認為是腫瘤復發(fā)轉(zhuǎn)移以及藥物抗性的重要原因,因此我們進一步檢測美登木素對鼻咽癌細胞腫瘤球形成能力的影響。我們發(fā)現(xiàn)腫瘤球大小以及數(shù)目受到美登木素的顯著抑制,蛋白檢測同樣表明腫瘤干細胞相關(guān)標記物受到顯著抑制。綜合以上實驗結(jié)果我們認為,美登木素通過抑制鼻咽癌細胞EMT以抑制其侵襲遷移以及腫瘤干細胞的增殖。基于EMT在美登木素對鼻咽癌生物學功能的影響中居于核心地位,隨后我們進一步利用生物信息學分析了美登木素抑制鼻咽癌細胞EMT的機制。結(jié)果表明,美登木素可能通過SMAD2抑制鼻咽癌細胞的EMT。綜合以上結(jié)果,美登木素通過抑制鼻咽癌細胞EMT的發(fā)生,進而抑制其侵襲遷移和腫瘤干細胞增殖。其機制可能是通過影響SMAD2信號通路從而抑制鼻咽癌細胞的EMT�；诖�,我們認為美登木素可以作為潛在的治療發(fā)生局部復發(fā)和遠處轉(zhuǎn)移鼻咽癌患者的有效化療藥物。而將美登木素與相關(guān)抗體結(jié)合將更加有效的治療該類鼻咽癌患者。
[Abstract]:Nasopharyngeal carcinoma (NPC) is a malignant tumor in Guangdong, especially in Guangdong. For patients with early and local advanced nasopharyngeal carcinoma, the main treatment mode is chemotherapy or radiotherapy and chemotherapy, and the 5-year survival rate of these patients can reach 85%. However, some patient continue to fail due to distant metastasis and local recurrence. The platinum-containing dual-drug chemotherapy is a standard treatment for the treatment of local recurrence and distant metastasis of nasopharyngeal carcinoma. But the scheme has the limitations of short response duration, more complications and the like. Therefore, finding new treatment methods and drugs to reduce the complications and to improve the survival of patients with local recurrence and distant metastasis is still a clinical problem to be solved. With the improvement of the anti-drug-drug conjugate (ADC) technique, a group of drugs with strong anti-tumor effect, but with a certain toxicity, has been re-paid attention. Medenwood is the most widely used cytotoxic drug in antibody-drug conjugates. In this study, we will explore the effect of medenwood on the biological function of nasopharyngeal carcinoma. First of all, we analyzed the gene chips associated with metaxin by bioinformatics, which contains breast cancer tissues treated with T-DM1 (trastuzumab-emtansine, trastuzumab-metaxin-coupled), and trastuzumab-trastuzumab, respectively. By comparing two groups of different chips, we can conclude that the maidenwood derivative DM1 can affect the expression of the genes of the breast cancer cells. Finally, we found that DM1 could significantly inhibit the expression of EMT-related genes in breast cancer cells. The correlation study also shows that the metaxin can inhibit the occurrence of the breast cancer cell line EMT. Based on this, we further explore whether the metaxin inhibits the occurrence of EMT in the nasopharyngeal carcinoma cell. By means of MTT, plate clone formation experiment, cycle experiment and the detection of the related cycle protein, the subcutaneous tumorigenesis of the nude mice, we preliminarily confirmed that the metaxin significantly inhibited the proliferation ability of the inside and outside of the cell body of the nasopharyngeal carcinoma. Then, through the scratch test, Transwell and Bodyn, we confirm that the metaxin can significantly inhibit the invasion and migration ability of the nasopharyngeal carcinoma cell line. Thus, the phenotype of EMT was significantly affected by the metaxin. Thus, the change of EMT-related protein was further detected by Western blot, and the marker E-cadherin associated with the epithelial property was found to be significantly higher, and the marker N-cadherin and the vimentin marker associated with the properties of the mesenchymal cells were reduced. Thus, the EMT is inhibited and the invasion and migration ability of the cells is reduced under the action of medename. The tumor stem cells are considered to be an important cause of the metastasis of the tumor and the resistance of the drug, so we can further examine the effect of the metaxus on the formation of the tumor ball of the nasopharyngeal carcinoma cell. We found that the size of the tumor ball and the number of tumor cells are significantly inhibited by the metaxin, and the protein detection also indicates that the tumor stem cell-related marker is significantly inhibited. Taken together with the above experimental results, we believe that the metaxin inhibits the invasion and migration of the nasopharyngeal carcinoma cell EMT and the proliferation of the tumor stem cells. EMT, based on EMT, is at the core of the biological function of nasopharyngeal carcinoma, and then we use bioinformatics to analyze the mechanism of MEMT in nasopharyngeal carcinoma. The results show that MAD2 can inhibit the EMT of nasopharyngeal carcinoma cells through SMAD2. Based on the above results, the metaxin inhibits the occurrence of the EMT in the nasopharyngeal carcinoma cell, and further inhibits the invasion and migration of the nasopharyngeal carcinoma cell EMT and the proliferation of the tumor stem cells. The mechanism may be to suppress the EMT of the nasopharyngeal carcinoma cell by affecting the SMAD2 signal path. Based on this, we believe that metaxin can be used as a potential treatment for local recurrence and distant metastasis of nasopharyngeal carcinoma. And the combination of the metaxin with the relevant antibody will be more effective in the treatment of the patients with this type of nasopharyngeal carcinoma.
【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.63
本文編號：2491711
[Abstract]:Nasopharyngeal carcinoma (NPC) is a malignant tumor in Guangdong, especially in Guangdong. For patients with early and local advanced nasopharyngeal carcinoma, the main treatment mode is chemotherapy or radiotherapy and chemotherapy, and the 5-year survival rate of these patients can reach 85%. However, some patient continue to fail due to distant metastasis and local recurrence. The platinum-containing dual-drug chemotherapy is a standard treatment for the treatment of local recurrence and distant metastasis of nasopharyngeal carcinoma. But the scheme has the limitations of short response duration, more complications and the like. Therefore, finding new treatment methods and drugs to reduce the complications and to improve the survival of patients with local recurrence and distant metastasis is still a clinical problem to be solved. With the improvement of the anti-drug-drug conjugate (ADC) technique, a group of drugs with strong anti-tumor effect, but with a certain toxicity, has been re-paid attention. Medenwood is the most widely used cytotoxic drug in antibody-drug conjugates. In this study, we will explore the effect of medenwood on the biological function of nasopharyngeal carcinoma. First of all, we analyzed the gene chips associated with metaxin by bioinformatics, which contains breast cancer tissues treated with T-DM1 (trastuzumab-emtansine, trastuzumab-metaxin-coupled), and trastuzumab-trastuzumab, respectively. By comparing two groups of different chips, we can conclude that the maidenwood derivative DM1 can affect the expression of the genes of the breast cancer cells. Finally, we found that DM1 could significantly inhibit the expression of EMT-related genes in breast cancer cells. The correlation study also shows that the metaxin can inhibit the occurrence of the breast cancer cell line EMT. Based on this, we further explore whether the metaxin inhibits the occurrence of EMT in the nasopharyngeal carcinoma cell. By means of MTT, plate clone formation experiment, cycle experiment and the detection of the related cycle protein, the subcutaneous tumorigenesis of the nude mice, we preliminarily confirmed that the metaxin significantly inhibited the proliferation ability of the inside and outside of the cell body of the nasopharyngeal carcinoma. Then, through the scratch test, Transwell and Bodyn, we confirm that the metaxin can significantly inhibit the invasion and migration ability of the nasopharyngeal carcinoma cell line. Thus, the phenotype of EMT was significantly affected by the metaxin. Thus, the change of EMT-related protein was further detected by Western blot, and the marker E-cadherin associated with the epithelial property was found to be significantly higher, and the marker N-cadherin and the vimentin marker associated with the properties of the mesenchymal cells were reduced. Thus, the EMT is inhibited and the invasion and migration ability of the cells is reduced under the action of medename. The tumor stem cells are considered to be an important cause of the metastasis of the tumor and the resistance of the drug, so we can further examine the effect of the metaxus on the formation of the tumor ball of the nasopharyngeal carcinoma cell. We found that the size of the tumor ball and the number of tumor cells are significantly inhibited by the metaxin, and the protein detection also indicates that the tumor stem cell-related marker is significantly inhibited. Taken together with the above experimental results, we believe that the metaxin inhibits the invasion and migration of the nasopharyngeal carcinoma cell EMT and the proliferation of the tumor stem cells. EMT, based on EMT, is at the core of the biological function of nasopharyngeal carcinoma, and then we use bioinformatics to analyze the mechanism of MEMT in nasopharyngeal carcinoma. The results show that MAD2 can inhibit the EMT of nasopharyngeal carcinoma cells through SMAD2. Based on the above results, the metaxin inhibits the occurrence of the EMT in the nasopharyngeal carcinoma cell, and further inhibits the invasion and migration of the nasopharyngeal carcinoma cell EMT and the proliferation of the tumor stem cells. The mechanism may be to suppress the EMT of the nasopharyngeal carcinoma cell by affecting the SMAD2 signal path. Based on this, we believe that metaxin can be used as a potential treatment for local recurrence and distant metastasis of nasopharyngeal carcinoma. And the combination of the metaxin with the relevant antibody will be more effective in the treatment of the patients with this type of nasopharyngeal carcinoma.
【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.63
【參考文獻】
相關(guān)期刊論文 前1條
1 白東魯;植物抗癌成份美登素的全合成[J];中國科學院院刊;1987年02期
，本文編號：2491711
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