【作者】 刘斌；

【导师】 乔士兴；

【作者基本信息】 吉林大学 ， 外科学， 2014， 博士

【摘要】 结直肠癌（colorectal cancer, CRC）是比较常见的消化道恶性肿瘤，它的发病率仅次于胃癌和食道癌，高居全球恶性肿瘤的第四位，每年新增患者约100万人，每年因CRC死亡的人数约50万人。近年来，CRC的发病率正在逐渐增加。同时患者的发病年龄也趋于年轻化。研究显示，结直肠癌的好发部位多为直肠和直肠、乙状结直肠的交界处，占60%左右，发病年龄多在60~70岁之间，50岁以下者还不到20%。CRC前期治疗主要是以手术治疗为主，晚期则是以放化疗为主。据报道，CRC的死亡原因主要由于肿瘤的复发和转移。已有研究表明，HMGA1参与了许多和肿瘤相关基因的转录和调控的过程。另有研究发现，HMGA1在癌细胞中的含量极其丰富，说明HMGA1可能改变染色质的结构。目前，HMGA1在CRC中的高表达具体机制尚未明确。因此，探讨HMGA1在CRC中发病机制中的作用对于临床研究具有重要的现实意义。近年研究表明，S100A3作为S100蛋白家族中重要的癌症因子，已被众多学者公认和广泛的研究。然而，是否S100A3在结直肠癌中发挥着同样重要的作用呢？如果S100A3在结直肠癌中发挥作用，那么与HMGA1蛋白的关系如何呢?据国内文献报道称，斑蝥是昆虫大斑蝥或者小斑蝥的黄黑色干燥体,拥有攻毒蚀疮、具有逐痪散结的独特功效,人类应用斑蝥治疗癌症疾病已有200多年的发展历史。斑蝥可通过抑制癌细胞对氨基酸的摄取，从而抑制蛋白质的合成，同时刺激淋巴细胞、巨噬细胞及多形核细胞产生白介素，从而提高机体免疫力，增加机体抵抗力，加大对肿瘤细胞的杀伤力而达到治疗的目的；斑蝥抗癌机理则是抑制癌细胞DNA以及蛋白质合成，控制癌细胞线粒体的呼吸和酶的活性。斑蝥素在抑制肿瘤的同时，不降低外周血中的白细胞数量，对机体没有显著的免疫抑制作用，这在抗肿瘤药物中是很少见的，因此备受人们关注。近年来，随着抗肿瘤中药优势的凸显，人们对斑蝥素及它的衍生物的研究越来越多，其中对斑蝥素，斑蝥素酸钠及去甲斑蝥素的研究最多，迄今为止，国内外对于结直肠癌的抑制作用尚未明确。那么，斑蝥是否可以降低结直肠癌中S100A3和HMGA1的表达，进而起到与化疗药相似的治疗效果呢？本实验将对S100A3蛋白和HMGA1蛋白可能的相互关系进行研究；另外，探讨S100A3和HMGA1蛋白在结直肠癌组织发病机制中的作用以及斑蝥干预后对S100A3蛋白和HMGA1蛋白表达的影响，从而揭示S100A3蛋白和HMGA1蛋白在结直肠癌中的中作用以及斑蝥对结直肠癌干预后的影响，为今后防治结直肠癌提供新的依据。更多还原

【Abstract】 CRC is a kind of malignant lesions of colorectal epithelium in a variety ofenvironmental or genetic factors. It is one of the common malignant lesions. Globalannual new cases of CRC are about100million people, accounting for10%of allmalignant tumors to15%.High expression of HMGA1can be induced malignant transformation of normalcells. The amount of HMGA1expression is biggest in embryonic development. Itsexpression reduced in fully differentiated mature cells or was not expressed. Thereaseach between S100A3and tumor was few compared to other members of thefamily of calcium-binding S100A.The affinity of S100A3can be considered as azinc ion signaling proteins. S100A3expression may be involved in malignanttransformation in the absence of early breast cells. Whether S100A3and HMGA1play an important role in CRC?Cantharides are commonly known as Meloidae spot lytta insects. Cantharidinatewas a special effect for primary liver cancer. Cantharidinate was used more and morewidely. It was also gradually in-depth study. In recent years, study of cantharidinatemainly focused on the synthesis, biological functions, toxicity and physiologicalaspects in the United States and European countries. Anticancer mechanisms,pharmacology and applied research of cantharidinate were more in our country. Butthe cantharidinate for prevention and treatment of CRC had not been reported.This study investigated the preventive and therapeutic effects of cantharidinate onhuman colorectal cancer both in vivo and in vitro. In addition, the mechanismsinvolved in origination and development of renal interstitial fibrosis also beenapproached. The main research contents is composed of①the relationship ofHMGA1and S100A3may be linked and investigated the role of HMGA1and S100A3in human CRC②the role of cantharidinate are observed for CRC③Theinhibition role and effect of cantharidinate are investigated for HMGA1and S100A3gene in human colorectal cancer.④The effect of cantharidinate is analyzed byRaman spectra.Main results were as follows:1. The protein expression of HMGA1and S100A3was significantly increasedin cancer tissues compared with the adjacent tissues.2. The expression of S100A3and HMGA1may be correlated in CRC.3. Immunohistochemistry and Western blot results confirmed that theexpression of HMGA1and S100A3was significantly increased in cancer tissuecompared with adjacent tissues (P <0.05and P <0.01).4. After intervention of cantharidinate, Cell cycle occurs more obvious changesin colorectal cancer.The expression of HMGA1and S100A3decreased gradually(P<0.05). Its protein structure also improved after intervention of cantharidinatethrough Raman spectra.Conclusions were as follows:1. S100A3and HMGA1play an important role in colorectal cancer.2. Cantharidinate is similar with chemotherapy, it can reduce the gene andprotein expression of S100A3and HMGA1in tumor cells and improve cycle.更多还原