本文主要研究内容
作者梁欣欣（2019）在《重组地皮菜水应激蛋白抗结肠癌的分子靶点及活性片段研究》一文中研究指出：结肠癌是全球最常见的恶性肿瘤之一,2018年最新数据表明,在全球范围内结肠癌的发病率位于第三位,结肠癌的死亡率位于第二位。结肠癌传统的治疗方式以手术结合术后化疗为主,但其缺陷是易复发且化疗药物毒副作用大,这也是造成结肠癌患者死亡的主要原因。随着结肠癌治疗的研究,靶向治疗以其不损害正常细胞的优势变成结肠癌治疗的一个新趋势。近年来结肠癌治疗的有效靶点不断被发现,因此,从天然产物中筛选针对这些靶点的无毒副作用的靶向药物成为国内外科学家的研究热点。地皮菜又名地耳、地木耳,学名为Nostoc commune Vauch.,是一种固氮蓝绿色藻类。地皮菜营养成份丰富,包括蛋白质、脂类、维生素和磷、锌、钙等矿物质。地皮菜可作为一种美食,能够清热去火、补充营养;也可做药用,有降低血压血脂,滋养肝肾等功能,而且对老年痴呆症有疗效。本课题组前期研究表明,从地皮菜中提取的水应激蛋白WSP1在体内和体外具有靶向抗结肠癌的效应,通过基因工程技术体外重组表达的Re-WSP1,也具有显著的抗结肠癌效应,但Re-WSP1抗结肠癌效应的分子靶点及其发挥抗结肠癌效应的活性片段尚不清楚。因此,本研究主要探究了重组地皮菜水应激蛋白Re-WSP1抗结肠癌的活性片段及其分子靶点,进一步阐明其发挥抗结肠癌作用的分子机制,包含以下三个方面内容:1、Re-WSP1对结肠癌细胞中Wnt/β-catenin信号通路影响的研究。Western blot检测了Re-WSP1处理后,结肠癌细胞中Wnt/β-catenin信号通路相关蛋白的表达情况,即Re-WSP1对Wnt/β-catenin信号通路的影响。结果表明:Re-WSP1处理后,结肠癌细胞中β-catenin、Dvl、cyclin D1的表达显著降低。这说明Re-WSP1处理后,在结肠癌细胞中异常激活的Wnt/β-catenin信号通路被抑制。2、Re-WSP1抗结肠癌效应的分子靶点的研究。通过免疫荧光技术发现Re-WSP1作用于结肠癌细胞的表面;通过His-Pull down和Western blot发现,Re-WSP1与FZD1之间存在相互作用。抗体封堵进一步验证,结果表明:Re-WSP1具有显著抗结肠癌效应,而加入FZD1抗体后,Re-WSP1对结肠癌细胞的抑制作用有明显的逆转现象。以上结果表明FZD1为Re-WSP1发挥抗结肠癌效应的作用靶点。3、Re-WSP1抗结肠癌活性片段的筛选。分析预测Re-WSP1的序列和功能结构域,根据预测结果构建Re-WSP1的截短体WSP1-1、WSP1-2和WSP1-3,并分别进行外源表达。通过MTT检测三个截短体片段对于结肠癌细胞增殖的影响,结果显示WSP1-1的活性明显高于其它两个截短体;进一步通过细胞流式检测了WSP1-1对于结肠癌细胞凋亡的影响,结果表明WSP1-1能够促进结肠癌细胞的凋亡。以上结果表明WSP1-1为Re-WSP1发挥抗结肠癌功能的活性片段。以上研究表明,Re-WSP1通过抑制Wnt/β-catenin信号通路而发挥抗结肠癌效应;通过进一步对于作用靶点的研究,发现Re-WSP1作用于Wnt/β-catenin信号通路中的七次跨膜蛋白受体FZD1,即Re-WSP1通过靶向FZD1抑制了Wnt/β-catenin信号通路从而发挥抗结肠癌效应。通过基因工程技术,明确了Re-WSP1中发挥抗结肠癌效应的活性片段为截短体WSP1-1。本研究通过探究Re-WSP1发挥抗结肠癌效应的分子机制,为地皮菜水应激蛋白对结肠癌的治疗作用及有效的生物靶点提供了坚实的理论支持,为其进一步的临床应用提供了可能。
Abstract
jie chang ai shi quan qiu zui chang jian de e xing zhong liu zhi yi ,2018nian zui xin shu ju biao ming ,zai quan qiu fan wei nei jie chang ai de fa bing lv wei yu di san wei ,jie chang ai de si wang lv wei yu di er wei 。jie chang ai chuan tong de zhi liao fang shi yi shou shu jie ge shu hou hua liao wei zhu ,dan ji que xian shi yi fu fa ju hua liao yao wu du fu zuo yong da ,zhe ye shi zao cheng jie chang ai huan zhe si wang de zhu yao yuan yin 。sui zhao jie chang ai zhi liao de yan jiu ,ba xiang zhi liao yi ji bu sun hai zheng chang xi bao de you shi bian cheng jie chang ai zhi liao de yi ge xin qu shi 。jin nian lai jie chang ai zhi liao de you xiao ba dian bu duan bei fa xian ,yin ci ,cong tian ran chan wu zhong shai shua zhen dui zhe xie ba dian de mo du fu zuo yong de ba xiang yao wu cheng wei guo nei wai ke xue jia de yan jiu re dian 。de pi cai you ming de er 、de mu er ,xue ming wei Nostoc commune Vauch.,shi yi chong gu dan lan lu se zao lei 。de pi cai ying yang cheng fen feng fu ,bao gua dan bai zhi 、zhi lei 、wei sheng su he lin 、xin 、gai deng kuang wu zhi 。de pi cai ke zuo wei yi chong mei shi ,neng gou qing re qu huo 、bu chong ying yang ;ye ke zuo yao yong ,you jiang di xie ya xie zhi ,zi yang gan shen deng gong neng ,er ju dui lao nian chi dai zheng you liao xiao 。ben ke ti zu qian ji yan jiu biao ming ,cong de pi cai zhong di qu de shui ying ji dan bai WSP1zai ti nei he ti wai ju you ba xiang kang jie chang ai de xiao ying ,tong guo ji yin gong cheng ji shu ti wai chong zu biao da de Re-WSP1,ye ju you xian zhe de kang jie chang ai xiao ying ,dan Re-WSP1kang jie chang ai xiao ying de fen zi ba dian ji ji fa hui kang jie chang ai xiao ying de huo xing pian duan shang bu qing chu 。yin ci ,ben yan jiu zhu yao tan jiu le chong zu de pi cai shui ying ji dan bai Re-WSP1kang jie chang ai de huo xing pian duan ji ji fen zi ba dian ,jin yi bu chan ming ji fa hui kang jie chang ai zuo yong de fen zi ji zhi ,bao han yi xia san ge fang mian nei rong :1、Re-WSP1dui jie chang ai xi bao zhong Wnt/β-cateninxin hao tong lu ying xiang de yan jiu 。Western blotjian ce le Re-WSP1chu li hou ,jie chang ai xi bao zhong Wnt/β-cateninxin hao tong lu xiang guan dan bai de biao da qing kuang ,ji Re-WSP1dui Wnt/β-cateninxin hao tong lu de ying xiang 。jie guo biao ming :Re-WSP1chu li hou ,jie chang ai xi bao zhong β-catenin、Dvl、cyclin D1de biao da xian zhe jiang di 。zhe shui ming Re-WSP1chu li hou ,zai jie chang ai xi bao zhong yi chang ji huo de Wnt/β-cateninxin hao tong lu bei yi zhi 。2、Re-WSP1kang jie chang ai xiao ying de fen zi ba dian de yan jiu 。tong guo mian yi ying guang ji shu fa xian Re-WSP1zuo yong yu jie chang ai xi bao de biao mian ;tong guo His-Pull downhe Western blotfa xian ,Re-WSP1yu FZD1zhi jian cun zai xiang hu zuo yong 。kang ti feng du jin yi bu yan zheng ,jie guo biao ming :Re-WSP1ju you xian zhe kang jie chang ai xiao ying ,er jia ru FZD1kang ti hou ,Re-WSP1dui jie chang ai xi bao de yi zhi zuo yong you ming xian de ni zhuai xian xiang 。yi shang jie guo biao ming FZD1wei Re-WSP1fa hui kang jie chang ai xiao ying de zuo yong ba dian 。3、Re-WSP1kang jie chang ai huo xing pian duan de shai shua 。fen xi yu ce Re-WSP1de xu lie he gong neng jie gou yu ,gen ju yu ce jie guo gou jian Re-WSP1de jie duan ti WSP1-1、WSP1-2he WSP1-3,bing fen bie jin hang wai yuan biao da 。tong guo MTTjian ce san ge jie duan ti pian duan dui yu jie chang ai xi bao zeng shi de ying xiang ,jie guo xian shi WSP1-1de huo xing ming xian gao yu ji ta liang ge jie duan ti ;jin yi bu tong guo xi bao liu shi jian ce le WSP1-1dui yu jie chang ai xi bao diao wang de ying xiang ,jie guo biao ming WSP1-1neng gou cu jin jie chang ai xi bao de diao wang 。yi shang jie guo biao ming WSP1-1wei Re-WSP1fa hui kang jie chang ai gong neng de huo xing pian duan 。yi shang yan jiu biao ming ,Re-WSP1tong guo yi zhi Wnt/β-cateninxin hao tong lu er fa hui kang jie chang ai xiao ying ;tong guo jin yi bu dui yu zuo yong ba dian de yan jiu ,fa xian Re-WSP1zuo yong yu Wnt/β-cateninxin hao tong lu zhong de qi ci kua mo dan bai shou ti FZD1,ji Re-WSP1tong guo ba xiang FZD1yi zhi le Wnt/β-cateninxin hao tong lu cong er fa hui kang jie chang ai xiao ying 。tong guo ji yin gong cheng ji shu ,ming que le Re-WSP1zhong fa hui kang jie chang ai xiao ying de huo xing pian duan wei jie duan ti WSP1-1。ben yan jiu tong guo tan jiu Re-WSP1fa hui kang jie chang ai xiao ying de fen zi ji zhi ,wei de pi cai shui ying ji dan bai dui jie chang ai de zhi liao zuo yong ji you xiao de sheng wu ba dian di gong le jian shi de li lun zhi chi ,wei ji jin yi bu de lin chuang ying yong di gong le ke neng 。
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论文作者分别是来自山西大学的梁欣欣，发表于刊物山西大学2019-11-12论文，是一篇关于结肠癌论文,山西大学2019-11-12论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自山西大学2019-11-12论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
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