丁酸钠诱导结肠癌Lovo细胞凋亡及其对p53靶基因的调控.docVIP

丁酸钠诱导结肠癌Lovo细胞凋亡及其对p53靶基因的调控 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：丁酸钠诱导结肠癌Lovo细胞凋亡及其对p53靶基因的调控 1 1 材料与方法 3 2 结果 4 3 讨论 6 文2：Bax蛋白在丁酸钠诱导食管癌细胞凋亡中的表达 8 1材料和方法 9 2 结果 10 2．1 倒置显微镜下观察结果 10 2．2 免疫组化检测凋亡相关基因bax蛋白的表达 10 3 讨论 10 参考文摘引言： 11 原创性声明（模板） 13 文章致谢（模板） 13 正文 丁酸钠诱导结肠癌Lovo细胞凋亡及其对p53靶基因的调控 文1：丁酸钠诱导结肠癌Lovo细胞凋亡及其对p53靶基因的调控 【Abstract】 Objective To observe the regulation of sodium butyrate on the three main target genes of p53 ,including p21waf1,Bax and Gadd45,and to explore the molecule mechanism of its inhibiting proliferation and inducing apoptosis of Lovo colon cancer The colon cancer cell line Lovo was cultivated in different concentratio of NaBT which concentratio were /L， mmol/L， mmol/L，10 mmol/L respectively. MTT was used to detect the cellular proliferation. The apoptosis was examined by both flow cytometry and Annexin VFITC/PI double tagging. RTPCR and Western blotting were used to study the expression of p21waf1,Bax,and Gadd45 messenger ribonuclcic acid(mRNA) and that of the Sodium butyrate inhibited proliferation， induced apoptosis，and blocked Lovo cell mainly in G2 phase in a time and dosedependant manner. The expression of p21waf1, Bax and Gadd45 gene was all enhanced after being treated with sodium butyrate both at mRNA and protein levels, and the expression of Gadd45 was the most obvious of Sodium butyrate above mmol/L may inhibit proliferation, induce apoptosis and blocked Lovo cell mainly in G2 phase in a time and dosedependant manner. Maybe the effect associated with its upregulating the expressio of p21waf1,Bax and Gadd45 within Lovo cell. 【Key words】 Lovo colorectal cancer cell line, sodium butyrate, cell apoptosis ,p53 target genes 丁酸钠是膳食纤维在肠道厌氧菌作用下产生的一种四碳短链脂肪酸（丁酸）的钠盐。研究发现，它可以对许多体外培养的哺乳动物细胞产生多种生物学作用。本研究选择携带野生型p53基因的结肠癌Lovo细胞株为研究对象，通过分析丁酸钠对Lovo细胞p53基因的三个主要下游基因，即p21waf1、Bax和Gadd45的调控，旨在进一步探讨其抑制细胞增生、诱导细胞凋亡的分子机制。 1 材料与方法 细胞培养及试剂 结肠癌Lovo细胞株购于山东省医学科学院。常规培养于含10%小牛血清的RPMI 1640培养液，37℃、5% CO2 ，饱和湿度的培养箱中。丁酸钠（Si