摘要肿瘤是全球发病率最高的疾病之一。原发性肝癌，也成为肝细胞性肝癌（hepatocellular carcinoma ,HCC），是世界上流行的恶性肿瘤之一。目前，癌症的治疗主要是放疗、化疗和手术，预后较差，且病人需要承受较大的痛苦。因此，我们迫切需要寻求一种能够替代化疗、放疗，有效控制癌症的方法。中药是我国传统中医特有药物，它的许多疗效已经被现代科学所证实，是治疗癌症的一种新机会。姜黄素是中药的一种，是一种多酚类物质，从姜的根、茎中获得，长期以来被用于治疗几种慢性疾病。研究表明，姜黄素具有抗肿瘤、抗氧化、抗炎等性质。75272

本实验前期利用姜黄素处理HepG-2细胞，发现姜黄素有抑制细胞增殖、促进细胞凋亡的作用。通过基因筛选发现姜黄素可通过提高XAF1基因表达而抑制HepG-2细胞增殖。本实验旨在探讨姜黄素对HepG-2细胞中XAF1基因表达的影响，并构建XAF1 基因反义干扰表达载体，建立XAF1 基因沉默的稳细胞系，将为进一步深入研究XAF1基因在姜黄素抑制HepG-2细胞增殖中的作用奠定基础。

Abstract Cancer is one of the highest incidence of disease in the world。 Primary hepatocellular carcinoma (HCC), is one of the malignant tumors in the world。 At present, the treatment of cancer is mainly radiotherapy, chemotherapy and surgery, the prognosis is poor, and patients need to bear greater pain。 Therefore, we urgently need to find a way to replace chemotherapy, radiotherapy, effective control of cancer。 Traditional Chinese medicine has a lot of efficacy has been confirmed by modern science, is a new opportunity for the treatment of cancer。 Curcumin is a kind of traditional Chinese medicine, is a kind of polyphenols, obtained from the root and stem ginger, for a long time is used to treat some chronic diseases。 The study showed that curcumin has the properties of anti tumor, anti oxidation, anti inflammation and so on。。

In the early stage of the experiment, we found that curcumin could inhibit the proliferation and promote the apoptosis of HepG-2 cells。 It was found that curcumin could inhibit the proliferation of HepG-2 cells by increasing the expression of XAF1 gene。 This experiment aims to investigate the effect of curcumin on the effect of XAF1 Gene Expression in HepG-2 cells and construction of antisense XAF1 Gene interference expression vector and establishment of XAF1 Gene silencing of stable cell lines for further study of XAF1 Gene in curcumin can inhibit the proliferation of HepG-2 cells in the role of foundation foundation。

毕业论文关键词：姜黄素；HepG-2细胞；XAF1基因；基因表达；反义干扰；载体构建

Key words:  curcumin; HepG-2 cell; XAF1 gene; gene expression; shRNA; vectorconstruction

目录

目录 3

引言 4

1 材料与方法 5

1。1  实验材料 5

1。2  培养方法 5

1。2。1 细胞培养 5

1。2。2 细胞处理 6

1。2。3 Trizol法提总RNA 6

1。2。4半定量RT-PCR检测XAF1基因mRNA表达情况 6

1。2。5 XAF1基因shRNA的设计与合成 6

1。2。6 XAF1基因shRNA表达载体pGreen-Puro-XAF1 shRNA的构建 7

1。2。7 构建的pGreen-Puro-XAF1 shRNA表达载体的鉴定 8

1。2。8 细胞转染及稳定干扰细胞系的建立