表皮葡萄球菌来源的脂肽衍生物调节黑色素瘤细胞增殖、分化与迁移的功能机制
发布时间：2019-03-26 19:46
【摘要】：黑色素瘤起源于黑色素细胞,是发病率最低然而致死率却最高的皮肤癌。由于传统的治疗方法如手术、化疗和放射治疗等不能解决黑色素瘤转移和复发问题,因此靶向治疗和免疫治疗成为主要研究方向。目前,临床应用的治疗黑色素瘤药物多为针对原癌基因如BRAF、MEK的抑制剂。但是长期服用抑制剂导致肿瘤的耐药性和机体副作用使得黑色素瘤治疗仍然存在局限性。因此,人们越来越关注从自然界寻找一些天然化合物应用于肿瘤治疗中。脂肽是微生物的次生代谢产物,具有广泛的生物活性,如消炎、杀菌、抗肿瘤等。表皮葡萄球菌是一类皮肤共生菌,我们实验室从其发酵液中提取出一种脂肽,能够诱导角质形成细胞分泌抗菌肽,具有杀菌活性,而且其同分异构体具有抑制皮肤伤口过度炎症,调节皮肤先天免疫应答的活性。本研究通过细胞周期流式分析和体外克隆形成实验证明了表皮葡萄球菌来源的脂肽衍生物LP79能够诱导黑色素瘤细胞B16F10停滞在细胞周期G1期,进而抑制其增殖。Transwell和伤口实验表明LP79能够显著性抑制B16F10细胞的迁移。而且,我们通过构建实验性肿瘤转移模型,发现LP79抑制小鼠黑色素瘤的肺部迁移。进一步研究表明,脂肽LP79能通过调节双重特异性磷酸酶MKP1进而抑制肿瘤细胞在上皮样细胞转化过程中的两个重要蛋白——N-钙黏蛋白和波形蛋白——的表达,从而抑制小鼠黑色素瘤的转移。另外,脂肽LP79能够通过诱导酪氨酸酶相关蛋白TRP1进而促进小鼠黑色素瘤细胞的分化。然而当TRP1基因沉默之后,LP79依然能够抑制B16F10细胞的迁移,表明二者并无直接相关性。此外,TLR2作为脂肽的受体,并不参与脂肽LP79对黑色素瘤细胞B16F10的抑制迁移功能。综上所述,本研究证明了表皮葡萄球菌来源的脂肽衍生物具有抑制黑色素瘤迁移的生物学活性。本研究进一步揭示了皮肤共生菌对宿主的益处,为治疗黑色素瘤的新药研发提出了独特的视角,也为治疗肿瘤提供了新的思路。
[Abstract]:Melanoma, originated from melanocytes, is the lowest incidence but the highest fatality rate of skin cancer. Because traditional treatment methods such as surgery chemotherapy and radiotherapy can not solve the problem of metastasis and recurrence of melanoma targeted therapy and immunotherapy have become the main research direction. At present, most of the drugs used in the treatment of melanoma are inhibitors of proto-oncogene such as BRAF,MEK. However, long-term use of inhibitors leads to tumor resistance and side effects, so melanoma treatment is still limited. As a result, more and more attention has been paid to finding natural compounds from nature for use in cancer therapy. Lipopeptide is the secondary metabolite of microorganism. It has a wide range of biological activities, such as anti-inflammatory, bactericidal, anti-tumor and so on. Staphylococcus epidermidis is a kind of symbiotic bacteria in skin. We extracted a lipopeptide from its fermentation broth, which can induce keratinocytes to secrete antibacterial peptides and have bactericidal activity. Moreover, the isomers can inhibit the excessive inflammation of skin wound and regulate the innate immune response of skin. In this study, we demonstrated that LP79, a lipopeptide derivative from Staphylococcus epidermidis, could induce B16F10 arrest in the G1 phase of melanoma cells by cell cycle flow analysis and clone formation in vitro. Transwell and wound experiments showed that LP79 could significantly inhibit the migration of B16F10 cells. Furthermore, we found that LP79 inhibits pulmonary migration of melanoma in mice by constructing experimental tumor metastasis model. Further studies showed that lipopeptide LP79 could inhibit the expression of two important proteins, N-cadherin and vimentin, during the transformation of epithelial-like cells by regulating the double specific phosphatase MKP1. Thus, the metastasis of melanoma in mice was inhibited. In addition, lipopeptide LP79 can promote the differentiation of mouse melanoma cells by inducing tyrosinase-associated protein TRP1. However, when TRP1 gene was silenced, LP79 could still inhibit the migration of B16F10 cells, indicating that there was no direct correlation between them. In addition, TLR2, as the receptor of lipopeptide, does not participate in the inhibitory effect of lipopeptide LP79 on B16F10 migration in melanoma cells. In conclusion, this study demonstrated that the lipopeptide derivatives derived from Staphylococcus epidermidis have the biological activity of inhibiting melanoma migration. This study further reveals the host benefits of skin symbiotic bacteria, provides a unique perspective for the research and development of new drugs for the treatment of melanoma, and also provides a new way of thinking for the treatment of tumors.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.5
本文编号：2447866
[Abstract]:Melanoma, originated from melanocytes, is the lowest incidence but the highest fatality rate of skin cancer. Because traditional treatment methods such as surgery chemotherapy and radiotherapy can not solve the problem of metastasis and recurrence of melanoma targeted therapy and immunotherapy have become the main research direction. At present, most of the drugs used in the treatment of melanoma are inhibitors of proto-oncogene such as BRAF,MEK. However, long-term use of inhibitors leads to tumor resistance and side effects, so melanoma treatment is still limited. As a result, more and more attention has been paid to finding natural compounds from nature for use in cancer therapy. Lipopeptide is the secondary metabolite of microorganism. It has a wide range of biological activities, such as anti-inflammatory, bactericidal, anti-tumor and so on. Staphylococcus epidermidis is a kind of symbiotic bacteria in skin. We extracted a lipopeptide from its fermentation broth, which can induce keratinocytes to secrete antibacterial peptides and have bactericidal activity. Moreover, the isomers can inhibit the excessive inflammation of skin wound and regulate the innate immune response of skin. In this study, we demonstrated that LP79, a lipopeptide derivative from Staphylococcus epidermidis, could induce B16F10 arrest in the G1 phase of melanoma cells by cell cycle flow analysis and clone formation in vitro. Transwell and wound experiments showed that LP79 could significantly inhibit the migration of B16F10 cells. Furthermore, we found that LP79 inhibits pulmonary migration of melanoma in mice by constructing experimental tumor metastasis model. Further studies showed that lipopeptide LP79 could inhibit the expression of two important proteins, N-cadherin and vimentin, during the transformation of epithelial-like cells by regulating the double specific phosphatase MKP1. Thus, the metastasis of melanoma in mice was inhibited. In addition, lipopeptide LP79 can promote the differentiation of mouse melanoma cells by inducing tyrosinase-associated protein TRP1. However, when TRP1 gene was silenced, LP79 could still inhibit the migration of B16F10 cells, indicating that there was no direct correlation between them. In addition, TLR2, as the receptor of lipopeptide, does not participate in the inhibitory effect of lipopeptide LP79 on B16F10 migration in melanoma cells. In conclusion, this study demonstrated that the lipopeptide derivatives derived from Staphylococcus epidermidis have the biological activity of inhibiting melanoma migration. This study further reveals the host benefits of skin symbiotic bacteria, provides a unique perspective for the research and development of new drugs for the treatment of melanoma, and also provides a new way of thinking for the treatment of tumors.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.5
【参考文献】
相关期刊论文 前1条
1 卢大用,曹静懿;肿瘤转移的实验病理学研究[J];河南医学研究;2000年03期
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