BCAT1抑制剂Gabapentin影响肝癌细胞增殖、迁移、凋亡及其机制的实验研究

《BCAT1抑制剂Gabapentin影响肝癌细胞增殖、迁移、凋亡及其机制的实验研究》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、分类号：R6单位代码：10343学号：151001201硕士学位论文论文题目:BCAT1抑制剂Gabapentin影响肝癌细胞增殖、迁移、凋亡及其机制的实验研究研究生姓名：赵炜嘉学科专业：外科学类型：学术型指导教师：张启瑜教授二O一八年五月温州医科大学硕士学位论文论文题目：BCAT1抑制剂Gabapentin影响肝癌细胞增殖、迁移、凋亡及其机制的实验研究答辩委员会主席：叶再元教授浙江省人民医院答辩委员会成员：马烽教授中国医学科学院周蒙滔教授温州医科大学附属第一医院论文答辩日期：2018年5月27日温州医科大学硕士学位论文目录一、缩略词

2、表„„„„„„„„„„„„„„„„„„„„„„„1二、摘要1、中文摘要„„„„„„„„„„„„„„„„„„„„„„„22、英文摘要„„„„„„„„„„„„„„„„„„„„„„„4三、正文1、前言„„„„„„„„„„„„„„„„„„„„„„„„„72、材料和方法„„„„„„„„„„„„„„„„„„„„„„93、实验结果„„„„„„„„„„„„„„„„„„„„„„„244、讨论„„„„„„„„„„„„„„„„„„„„„„„„„375、结论„„„„„„„„„„„„„„„„„„„„„„„„„406、参考文献„„„„„„„„„„„„„„„„„„

3、„„„„„41四、附录„„„„„„„„„„„„„„„„„„„„„„„„45五、致谢„„„„„„„„„„„„„„„„„„„„„„„„46六、综述„„„„„„„„„„„„„„„„„„„„„„„„47参考文献„„„„„„„„„„„„„„„„„„„„„„52七、独创性声明„„„„„„„„„„„„„„„„„„„„„56八、使用授权声明„„„„„„„„„„„„„„„„„„„„56温州医科大学硕士学位论文缩略词缩略词英文全称中文全称MTTMethylthiazolyldiphenyl-tetrazoliumbromide噻唑蓝ODopticalde

4、nsity光密度BCATBranched-chainaminoacidaminotransferase支链氨基酸转氨酶ddH2ODistillation-DistillationH2O双蒸水mMmmol/L微摩尔每升PIPropidiumiodide碘化丙啶FITCfluoresceinisothiocyanate异硫氰酸荧光素DEPCdiethylpyrocarbonate焦炭酸二乙酯RTReverseTranscription逆转录PCRPolymeraseChainReaction聚合酶链反应DNADeoxyriboNuclei

5、cAcid脱氧核糖核酸mRNAMessageRiboseNucleicAcid信使核糖核酸DMSODimethylsulfate二甲基亚砜EDTAEhylenediaminetetraaceticacid乙二胺四乙酸rpmRevolutionperminute每分钟转数FBSFetalBovineSerum胎牛血清DMEMdulbecco'smodifiedeaglemedium杜氏细胞培养基PBSphosphatebuffersaline磷酸盐缓冲液HCChepatocellularcarcinoma肝细胞性肝癌shRNAshort

6、hairpinRNA短发夹RNAGAPDHglyceraldehyde-3-phosphatedehydrogenase甘油醛-3-磷酸脱氢酶CDH11cadherin11钙黏附蛋白11FGF13fibroblastgrowthfactor13成纤维细胞生长因子131温州医科大学硕士学位论文BCAT1抑制剂Gabapentin影响肝癌细胞增殖、迁移、凋亡及其机制的实验研究目的：通过体外实验抑制肝癌细胞的支链氨基酸转氨酶1（BCAT1）表达观察BCAT1对肝癌细胞的增殖、迁移的影响，进一步使用BCAT1抑制剂Gabapentin处理肝癌

7、细胞，观察肝癌细胞增殖、迁移、凋亡、成瘤能力的变化以及使用RNA表达谱测序检测相关通路的基因表达的变化，从而进一步研究Gabapentin对肝癌细胞的影响，并对其可能的作用机制进行初步探讨。方法：体外培养人肝癌细胞株LM3，通过包含BCAT1干扰序列的shRNA慢病毒转染LM3细胞从而抑制LM3细胞的BCAT1表达，通过qRT-PCR和荧光显微镜检测评估转染效果，使用MTT法检测BCAT1抑制后肝癌细胞增殖能力的变化；使用细胞划痕实验检测BCAT1抑制后肝癌细胞迁移能力的变化。选取对数生长期多种人肝癌细胞株，予以不同药物浓度Gabap

8、entin（0mM，5mM,10mM,20mM,40mM，80mM）处理24h、48h，采用MTT法对Gabapentin处理后的肝癌细胞增殖情况的变化进行检测。运用Transwell迁移实验检测Gabapentin处理