瘦素和SIRT1对骨肉瘤细胞增殖、转移的影响及机制研究

发布时间：2018-05-30 00:10

本文选题：骨肉瘤 + Leptin　；参考：《河北师范大学》2016年博士论文

【摘要】：研究目的:骨肉瘤(Osteosarcoma,OS)是一种骨髓间充质来源的局部侵袭性的恶性肿瘤,易发生早期侵袭转移。OS常表现为局部浸润和早期肺转移,伴发转移的患者5年生存率仅为20%。近年来由于手术与新辅助化疗的应用延长了患者的生存期,改善了患者生存质量,但OS的整体治疗效果仍不理想。由于对多种化疗药物不敏感,顽固性的OS患者亟需更好的预后指标和更多的有效治疗靶点。瘦素(Leptin),一种由脂肪细胞分泌的内分泌激素,作为一个潜在的抗癌药物靶点已获得越来越多的关注。Sirutin-1(SIRT1)作为Leptin下游的靶点,是哺乳动物中重要的NAD+依赖性去乙酰化酶。本研究检测Leptin和SIRT1在人骨肉瘤组织中的表达;观察Leptin和SIRT1对骨肉瘤细胞增殖和转移的影响;探讨SIRT1介导Leptin促进骨肉瘤细胞侵袭转移的机制,并将数据与OS患者的临床病理资料进行了相关分析。本研究结果不仅具有理论价值,而且对于OS的临床治疗及预后判断也具有指导意义。研究方法:首先检测Leptin和SIRT1在人骨肉瘤组织中的表达,根据表达水平的差别结合临床随访资料,采用单因素及多因素方差分析,评估表达与肿瘤的不良临床特征及预后的关系。然后采用MTT、流式细胞仪、细胞划痕实验、Transwell实验、Westernblot等实验方法对骨肉瘤MG-63细胞的生长特性及形态学表现进行研究,以评估Leptin和SIRT1在骨肉瘤细胞中的表达及对其增殖转移侵袭能力的影响,通过抑制SIRT1表达对Leptin促进骨肉瘤细胞增殖侵袭转移的影响来印证二者在OS的相关性。研究结果:1、Leptin和SIRT1在OS中呈高表达。高表达的Leptin和SIRT1与不良临床特征显著相关。无论患者是否接受了新辅助化疗,Leptin与SIRT1的表达水平呈正相关,且二者均与远处转移和Enneking分期呈正相关。2、Leptin通过上调VEGF的表达促进了骨肉瘤细胞的血管生成能力,Leptin可促进骨肉瘤细胞的增殖和抗凋亡能力,并可以降低骨肉瘤细胞的粘附性和增强侵袭能力。3、SIRT1可能通过VEGF途径增强血管生成能力从而促进了肿瘤的局部侵袭、增殖和抗凋亡能力。4、Leptin可显著促进骨肉瘤细胞中SIRT1的表达,抑制骨肉瘤细胞中SIRT1的表达可有效抑制Leptin促进骨肉瘤细胞增殖侵袭转移能力。SIRT1可能通过促进骨肉瘤细胞中MMP2、MMP8及MMP9的表达促进骨肉瘤细胞的侵袭转移。研究结论:1、Leptin和SIRT1在OS中高表达。高表达的Leptin和SIRT1与不良临床特征显著相关。无论患者是否接受了新辅助化疗,Leptin与SIRT1的表达水平正相关,且二者均与远处转移和Enneking分期正相关。SIRT1可能在OS发生发展中发挥重要的作用。SIRT1的表达水平对于OS患者是一个重要的临床预后因子,也可作为OS患者治疗的新靶点。2、Leptin通过上调VEGF的表达促进了骨肉瘤细胞的血管生成能力,同时我们发现Leptin可促进骨肉瘤细胞的增殖和抗凋亡能力,并可以降低骨肉瘤细胞的粘附性和增强侵袭能力,这些研究结果进一步证实了Leptin在OS的转移进程中起到重要的作用,为早期控制OS的发展提供潜在诊断、治疗策略及数据支持。3、SIRT1可能通过VEGF途径增强血管生成能力从而促进了肿瘤的局部侵袭,并且促进了骨肉瘤细胞增殖和抗凋亡能力,提示SIRT1在OS的发生发展中可能发挥着重要的作用。4、Leptin可显著促进骨肉瘤细胞中SIRT1的表达,而且抑制骨肉瘤细胞中SIRT1可有效抑制Leptin促进骨肉瘤细胞增殖侵袭转移能力,提示SIRT1介导了Leptin促进骨肉瘤细胞增殖侵袭转移能力。而SIRT1则可能通过促进骨肉瘤细胞中MMP2、MMP8及MMP9的表达促进骨肉瘤细胞的侵袭转移。本课题通过探讨OS发生发展过程中Leptin与SIRT1的调控机制,明确了骨肉瘤细胞生物学性能调控的作用靶点及分子机制,SIRT1作为OS一个潜在的基因治疗新靶点,值得进一步深入研究。
[Abstract]:Objective: Osteosarcoma (OS) is a locally invasive malignant tumor of bone marrow mesenchymal origin. Early invasion and metastasis is prone to local invasion and early pulmonary metastasis. The 5 year survival rate of patients with metastasis is only 20%. in recent years, which has extended the patient's survival time due to the application of surgery and neoadjuvant chemotherapy. The quality of life of the patients is good, but the overall treatment effect of OS is still not ideal. Because of the insensitivity to a variety of chemotherapeutic drugs, the intractable OS patients urgently need better prognostic indicators and more effective therapeutic targets. Leptin (Leptin), an endocrine hormone secreted by adipocytes, has gained more and more potential as a potential anti-cancer drug target. More attention is paid to.Sirutin-1 (SIRT1) as the target of the lower Leptin. It is an important NAD+ dependent deacetylase in mammalian. This study detected the expression of Leptin and SIRT1 in human osteosarcoma and the effects of Leptin and SIRT1 on the proliferation and metastasis of osteosarcoma cells, and discussed SIRT1 mediated Leptin to promote the invasion and metastasis of osteosarcoma cells. The data are analyzed with the clinicopathological data of OS patients. The results of this study not only have theoretical value, but also provide guidance for the clinical treatment and prognosis of OS. Research methods: first, to detect the appearance of Leptin and SIRT1 in human osteosarcoma tissue, according to the difference of expression level combined with clinical follow-up A single factor and multiple factor variance analysis were used to evaluate the relationship between the expression and the adverse clinical features and prognosis of the tumor. Then the growth characteristics and morphological features of osteosarcoma MG-63 cells were studied by MTT, flow cytometry, cell scratch test, Transwell experiment and Westernblot, so as to evaluate Leptin and SIRT1 in bone. The expression of the sarcomarosarcoma cells and its effect on its proliferation and invasion ability by inhibiting the effect of SIRT1 expression on the proliferation and invasion of osteosarcoma cells by Leptin to confirm the correlation between the two in OS. 1, Leptin and SIRT1 are highly expressed in OS. The high expression of Leptin and SIRT1 is significantly related to adverse clinical characteristics. Whether or not new adjuvant chemotherapy is accepted, Leptin is positively correlated with the expression level of SIRT1, and all of the two are positively related to distant metastasis and Enneking staging. Leptin can promote the angiogenesis of osteosarcoma cells by up regulation of VEGF, Leptin can promote the proliferation and anti apoptosis ability of osteosarcoma cells, and can reduce osteosarcoma fine. Cell adhesion and enhanced invasiveness.3, SIRT1 may enhance angiogenesis by VEGF pathway, thus promoting local invasion, proliferation and anti apoptosis,.4. Leptin can significantly promote the expression of SIRT1 in osteosarcoma cells. Inhibition of the expression of SIRT1 in osteosarcoma cells can effectively inhibit the proliferation of osteosarcoma cells by Leptin. .SIRT1 may promote the invasion and metastasis of osteosarcoma cells by promoting the expression of MMP2, MMP8 and MMP9 in osteosarcoma cells. Conclusions: 1, Leptin and SIRT1 are highly expressed in OS. The high expression of Leptin and SIRT1 are significantly associated with adverse clinical features. No matter whether patients receive new adjuvant chemotherapy, Leptin, and SIRT1 expression levels Positive correlation and positive correlation with distant metastasis and Enneking staging of the two.SIRT1 may play an important role in the development of OS. The expression level of.SIRT1 is an important prognostic factor for OS patients, and can also be a new target for the treatment of OS patients,.2. Leptin promotes the angiogenesis of osteosarcoma cells by up regulating the expression of VEGF. At the same time, we found that Leptin can promote the proliferation and anti apoptotic ability of osteosarcoma cells, reduce the adhesion of osteosarcoma cells and enhance the invasiveness of osteosarcoma cells. These findings further confirm that Leptin plays an important role in the process of OS metastasis and provides a potential diagnosis, treatment strategy and number for the development of early controlled OS. According to the support of.3, SIRT1 may enhance angiogenesis by VEGF pathway, promote local invasion of tumor and promote the proliferation and anti apoptosis of osteosarcoma cells, suggesting that SIRT1 may play an important role in the development of OS. Leptin can significantly promote the expression of SIRT1 in bone sarcoma cells and inhibit osteosarcoma. SIRT1 can effectively inhibit the proliferation and invasion and metastasis of osteosarcoma cells by Leptin, suggesting that SIRT1 promotes the proliferation and invasion of osteosarcoma cells by Leptin. SIRT1 may promote the invasion and metastasis of osteosarcoma cells by promoting the expression of MMP2, MMP8 and MMP9 in osteosarcoma cells. This topic is to explore the occurrence and development of OS. The regulatory mechanism of Leptin and SIRT1 in the process has clearly defined the target and molecular mechanism of the biological performance regulation of osteosarcoma cells. As a potential new target of gene therapy for OS, SIRT1 is worthy of further study.
【学位授予单位】：河北师范大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R738.1

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