美登木素抑制鼻咽癌细胞上皮间充质转化及其机理研究
发布时间：2019-06-03 05:32
【摘要】：鼻咽癌(nasopharyngeal carcinoma,NPC)是高发于中国南方尤其广东地区的恶性肿瘤。针对早期和局部晚期鼻咽癌患者,目前主要的治疗方式是化疗或者放化疗,这类患者的5年生存率可达到85%。但是,仍然有部分患者由于发生远处转移和局部复发而导致治疗失败。含铂的双药化疗是治疗局部复发和远处转移鼻咽癌患者的标准治疗方案。但是该方案存在缓解持续时间短,并发症较多等局限。因此寻找新的治疗方法和药物以减少并发症并提高局部复发和远处转移鼻咽癌患者的生存期仍然是亟待解决的临床问题。随着抗体-药物偶联物(Antibody-drug conjugate,ADC)技术的完善,一批抗肿瘤作用强,但具有一定毒性的药物又重新得到了关注。美登木素是抗体-药物偶联物中应用最广泛的细胞毒性药物。在本研究中,我们将探讨美登木素对鼻咽癌生物学功能的影响。首先我们通过生物信息学分析了美登木素相关的基因芯片,该芯片包含分别经T-DM1(trastuzumab-emtansine,曲妥珠单抗-美登木素偶联药物)以及trastuzumab曲妥珠单抗处理的乳腺癌组织。通过比对两组不同的芯片,我们可以得出美登木素衍生物DM1会影响乳腺癌细胞哪些基因的表达。最终我们发现DM1可以显著抑制乳腺癌细胞中EMT相关基因的表达。相关研究亦表明,美登木素可以抑制乳腺癌细胞株EMT的发生。基于此,我们进一步探索美登木素是否抑制了鼻咽癌细胞EMT的发生。通过MTT、平板克隆形成实验、周期实验以及相关周期蛋白的检测、裸鼠皮下成瘤,我们初步证实了美登木素显著抑制鼻咽癌细胞体内外的增殖能力。随后,通过划痕实验、Transwell以及Boyden,我们证实美登木素可以显著抑制鼻咽癌细胞株侵袭迁移能力。因此,EMT的表型受到了美登木素的显著影响。由此,进一步通过Western blot检测EMT相关蛋白的变化,发现上皮性质相关的标记物E-cadherin显著升高,而间充质细胞性质相关的标记物N-cadherin以及Vimentin标记物降低。由此说明鼻咽癌细胞在美登木素作用下,EMT受到抑制并降低了细胞的侵袭迁移能力。肿瘤干细胞被认为是肿瘤复发转移以及药物抗性的重要原因,因此我们进一步检测美登木素对鼻咽癌细胞肿瘤球形成能力的影响。我们发现肿瘤球大小以及数目受到美登木素的显著抑制,蛋白检测同样表明肿瘤干细胞相关标记物受到显著抑制。综合以上实验结果我们认为,美登木素通过抑制鼻咽癌细胞EMT以抑制其侵袭迁移以及肿瘤干细胞的增殖。基于EMT在美登木素对鼻咽癌生物学功能的影响中居于核心地位,随后我们进一步利用生物信息学分析了美登木素抑制鼻咽癌细胞EMT的机制。结果表明,美登木素可能通过SMAD2抑制鼻咽癌细胞的EMT。综合以上结果,美登木素通过抑制鼻咽癌细胞EMT的发生,进而抑制其侵袭迁移和肿瘤干细胞增殖。其机制可能是通过影响SMAD2信号通路从而抑制鼻咽癌细胞的EMT。基于此,我们认为美登木素可以作为潜在的治疗发生局部复发和远处转移鼻咽癌患者的有效化疗药物。而将美登木素与相关抗体结合将更加有效的治疗该类鼻咽癌患者。
[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.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.63
本文编号：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.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.63
【参考文献】
相关期刊论文 前1条
1 白东鲁;植物抗癌成份美登素的全合成[J];中国科学院院刊;1987年02期
，本文编号：2491711
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2491711.html
