简介:【摘要】 目的:观察程序性细胞死亡蛋白 5( programmed cell death5,PDCD5)对沙利度胺诱导的 RPMI-8226细胞凋亡中的作用,并探讨其可能存在的机制,为临床应用 PDCD5蛋白辅助性治疗多发性骨髓瘤提供实验依据。 方法:以 RPMI-8226细胞作为研究对象,将实验分为空白对照组和实验组进行体外细胞培养,空白对照组即不加任何处理的细胞,实验组分为 4组,分别为单独予 20mg/l PDCD5蛋白处理的细胞组、单独予 20mg/l 沙利度胺处理细胞组、予 20mg/l沙利度胺加 10mg/L PDCD5蛋白处理细胞组、予 20mg/l沙利度胺加 20mg/L PDCD5蛋白处理细胞组。 MTT比色法测定各个实验组和对照组作用 24h、 48h、 72h后细胞生长的抑制作用;流式细胞仪检测各个实验组和对照组细胞凋亡情况的表达;运用 RT-PCR检测各组细胞对 Survivin及 bcl-2 mRNA表达的影响;运用 Western Blot检测各组细胞对 Survivin及 bcl-2蛋白的表达变化。 结论: 1、沙利度胺能有效的诱导 RPMI-8226细胞凋亡,且随着作用时间增加,凋亡作用增强。 2 、 单用沙利度胺或联合 PDCD5蛋白诱导肿瘤细胞凋亡的同时,均下调 survivin、 BCL-2基因表达。 3 、 PDCD5蛋白不能直接明显的诱导 RPMI-8226细胞凋亡,但与沙利度胺联合应用时具有协同效应,并更显著的促进凋亡。
简介:摘要目的研究肝母细胞瘤HUH-6株与正常肝LO2细胞株microRNA-21及其靶基因PDCD4和PTEN表达的差别;研究抑制microRNA-21表达后肝母细胞瘤HUH-6细胞株中microRNA-21及其靶基因PDCD4和PTEN的变化,以及反义抑制后HUH-6细胞凋亡、细胞增殖周期的变化。方法正常肝LO2细胞株与肝母细胞瘤HUH-6株作为研究对象,将肝母细胞瘤HUH-6细胞株分为3组,包括:空白对照组(正常培养的HUH-6细胞);microRNA-21抑制组(转染microRNA-21抑制序列的细胞);阴性对照组(转染无关序列的HUH-6细胞)。运用实时荧光定量PCR技术分析各组细胞中的microRNA-21的表达水平。运用流式细胞技术检测反义抑制microRNA-21后HUH-6细胞凋亡以及细胞增殖情况。用Westen-blot检测细胞中PDCD4和PTEN的表达情况。结果与正常肝LO2细胞株相比,肝母细胞瘤HUH-6株microRNA-21表达上调;反义抑制后HUH-6细胞中microRNA-21表达下降;反义抑制后HUH-6细胞增殖率下降,凋亡率增加;反义抑制后HUH-6细胞PDCD4和PTEN表达增加。结论本研究通过细胞实验,发现肝母细胞瘤瘤细胞中microRNA-21表达上调,并可以负性调节靶基因蛋白PDCD4及PTEN蛋白的表达;microRNA-21可能通过PDCD4及PTEN蛋白调节肝母细胞瘤的增殖与凋亡。本研究首次进行microRNA-21在肝母细胞瘤细胞中的表达研究,为microRNA-21及靶基因PDCD4及PTEN在肝母细胞瘤发病机制的研究奠定基础,在肝母细胞瘤的治疗方面具有光明的研究前景。
简介:[摘要]在设计工作中有时会遇到建筑物基础不在同一标高,存在高差,且工程要求先施工较浅基础,经过计算对建筑物之间的距离提出最小要求成为方案设计时结构工程师的必要工作,本文结合工程实例提出最小距离的确定过程。
简介:AbstractBackground:Epithelial to mesenchymal transition (EMT) is a key process in determining distant metastasis and intra-hepatic dissemination of hepatocellular carcinoma (HCC). Follistatin (FST) family members are considered to be an attractive therapeutic targets and prognostic indicators in cancers. As a derivative of FST, Follistatin Like 5 (FSTL5) may play a similar role in HCC cells. This study aimed to investigate the expression and function of FSTL5 in HCC and its role in EMT.Methods:FSTL5, E-cadherin and vimentin in HCC, and paracancerous tissues were detected by immunohistochemistry. Correlation of FSTL5 expression with overall survival was assessed. The proliferation and invasion of HCC cell lines SK-Hep1 and MHCC-LM3 were analyzed by cell counting kit-8 and Transwell assays. The expression of FSTL5, E-cadherin, and vimentin in HCC cells was examined by polymerase chain reaction and Western blot analysis. T-test was used to analyze the difference in proliferation and invasion ability between groups. The Spearman rank correlation test was used to detect the correlation between the expression of FSTL5 and E-cadherin or vimentin.Results:The expression of FSTL5 in HCC was lower than that in paracancerous tissues (9.97% vs. 82.55%, χ2 = 340.15, P < 0.001). Patients with high FSTL5 expression had a better prognosis (χ2= 8.22, P= 0.004) and smaller tumor diameter (χ2 = 45.52, P < 0.001), less lymph node metastasis (χ2 = 5.58, P= 0.02), earlier tumor node metastasis stage (χ2 = 11.29, P= 0.001), a reduced number of tumors (χ2 = 5.05, P= 0.02), lower alpha-fetoprotein value (χ2 = 24.36, P < 0.001), more probability of hepatitis carrying (χ2 = 40.9, P < 0.001), and better liver function grade (χ2 = 5.21, P = 0.02). Immunohistochemistry showed that FSTL5 expression in HCC tissues was positively correlated with E-cadherin expression (r = 0.38, P < 0.001) and negatively correlated with vimentin expression (r = -0.385, P < 0.001). Furthermore, over-expression of FSTL5 up-regulated the expression of E-cadherin and down-regulated the expression of vimentin in SK-Hep1 (negative control [NC] vs. FSTL5-interfering group [Lv-FSTL5]: E-cadherin [t= 45.03, P < 0.001], vimentin [t= 67, P < 0.001]) and MHCC-LM3 (NC vs. Lv-FSTL5: E-cadherin [t = 50, P < 0.001], vimentin [t = 72.75, P < 0.001]) cells at mRNA level. The same as protein level. In addition, the over-expression of FSTL5 inhibited the proliferation (NC vs. Lv-FSTL5: SK-Hep1, 3 d [t = 7.324, P = 0.018], 4 d [t = 6.23, P = 0.021], 5 d [t= 10.21, P= 0.003]; MHCC-LM3, 3 d [t= 4.32, P= 0.037], 4 d [t= 7.49, P= 0.012], 5 d [t= 9.3661, P = 0.009]) and invasion (NC vs. Lv-FSTL5: SK-Hep1, t= 21.57, P < 0.001; MHCC-LM3, t= 18.04, P < 0.001) of HCC cells.Conclusions:Down-regulation of FSTL5 may contribute to EMT of HCC, and FSTL5 is a potential target in the treatment of HCC.
简介:摘要第5版WHO乳腺肿瘤分类已于2019年底正式出版,本文就新版分类中的主要更新内容进行解读。内容主要涉及新版WHO乳腺肿瘤分类的总体情况,以及浸润性癌、非浸润性癌、上皮-肌上皮肿瘤和纤维上皮性肿瘤内容的更新。
简介:【摘 要】语音作为语言的一个重要组成部分,是发展语言技巧的基础以及提高语言整体水平的关键因素。提高语音教学的效率是发展学生自主学习能力的途径之一。本文通过一节语音课剖析当下语音教学中存在的问题,提出改进的建议,从收集材料,解读发音和学后巩固等方面入手,反思课堂实施,构建基于美好教学愿景下的小学英语语音课堂。