AIM:To investigate the role of transforming growth factor(TGF)-β-inducible early gene 1(TIEG1) in TGF-β-induced growth inhibition in hepatocellular carcinoma(HCC) cells.METHODS:Human hepatocyte and HCC cell lines wi...AIM:To investigate the role of transforming growth factor(TGF)-β-inducible early gene 1(TIEG1) in TGF-β-induced growth inhibition in hepatocellular carcinoma(HCC) cells.METHODS:Human hepatocyte and HCC cell lines with varied susceptibilities to TGF-β1 were tested by methylthiazoletetrazolium(MTT) assay.The expression changes of Smad2,Smad3,Smad4,Smad7,TIEG1 and TIEG2 gene following treatment with TGF-β1 in a TGF-β-sensitive hepatocyte cell line(MIHA),a TGF-β-sensitive hepatoma cell line(Hep3B) and two TGF-β-insensitive hepatoma cell lines(HepG2 and Bel7404) were examined.SiRNA targeting TIEG1 was transfected into Hep3B cells and the sensitivity of cells to TGF-β1 was examined.Overexpression of TIEG1 was induced by lentiviral-mediated transduction in TGF-β1-resistant hepatoma cell lines(Bel7404 and HepG2).MTT assay and 4',6-Diamidino-2-phenylindole staining were used to identify cell viability and apoptosis,respectively.The expression level of stathmin was measured by reverse transcriptase polymerase chain reaction and Western-blotting analysis,and stathmin promoter activity by TIEG1 was monitored by a luciferase reporter gene system.RESULTS:TIEG1 was significantly upregulated by TGF-β1 in the TGF-β1-sensitive HCC cell line,Hep3B,but not in the resistant cell lines.The suppression of TIEG1 by siRNAs decreased the sensitivity of Hep3B cells to TGF-β1,whereas the overexpression of TIEG1 mediated growth inhibition and apoptosis in TGF-β1-resistant HCC cell lines,which resembled those of TGF-β1-sensitive HCC cells treated with TGF-β1.Our data further suggested that stathmin was a direct target of TIEG1,as stathmin was signif icantly downregulated by TIEG1 overexpression,and stathmin promoter activity was inhibited by TIEG1 in a dose-dependent manner.CONCLUSION:Our data suggest that transactivation of TIEG1 conferred growth inhibition of TGF-β-susceptible human HCC cells.展开更多
目的探讨以RNA干扰(RNAinterference,RNAi)技术靶向stathmin和mdr1基因逆转卵巢癌细胞紫杉醇耐药的可行性。方法分别构建靶向stathmin和mdr1基因的质粒:pGU6-GFP-neo-ST-MN1和pGU6-GFP-neo-MDR1;将质粒转染到卵巢癌紫杉醇耐药细胞株SKOV...目的探讨以RNA干扰(RNAinterference,RNAi)技术靶向stathmin和mdr1基因逆转卵巢癌细胞紫杉醇耐药的可行性。方法分别构建靶向stathmin和mdr1基因的质粒:pGU6-GFP-neo-ST-MN1和pGU6-GFP-neo-MDR1;将质粒转染到卵巢癌紫杉醇耐药细胞株SKOV3/TAX。Real-ti me RT-PCR检测stathminm和mdr1的mRNA变化;Western blot检测其蛋白表达变化;荧光显微镜检测细胞凋亡情况;CCK-8法分析细胞对紫杉醇的敏感度。结果 Real-ti me RT-PCR及Western blot显示pGU6-GFP-neo-MDR1质粒对mdr1基因,pGU6-GFP-STMN1-294shRNA质粒对stathmin基因在mR-NA水平和蛋白水平均有明显抑制(P<0.05),荧光显微镜下共转染组细胞凋亡增多,CCK-8示共转染组逆转紫杉醇耐药效果最明显。结论体外RNAi可有效沉默卵巢癌紫杉醇耐药株SKOV3/TAX细胞内stathmin基因和mdr1基因,逆转其紫杉醇耐药。展开更多
目的检测鼻腔鼻窦内翻性乳头状瘤中上皮细胞膜蛋白1(EMP1)、环氧化酶2(COX-2)及Stathmin蛋白的表达情况,分析其与上皮化生程度的相关性。方法选取2015年1月~2016年3月在本院就诊的鼻腔鼻窦内翻性乳头状瘤患者28例作为实验组,20例肥厚性...目的检测鼻腔鼻窦内翻性乳头状瘤中上皮细胞膜蛋白1(EMP1)、环氧化酶2(COX-2)及Stathmin蛋白的表达情况,分析其与上皮化生程度的相关性。方法选取2015年1月~2016年3月在本院就诊的鼻腔鼻窦内翻性乳头状瘤患者28例作为实验组,20例肥厚性鼻炎患者的下鼻甲黏膜组织作为对照组,采用免疫组化技术和q RT-PCR检测鼻腔鼻窦内翻性乳头状瘤、下鼻甲黏膜组织EMP1、COX-2以及Stathmin的表达情况,并比较三种因子在不同组织中的表达情况。结果实验组EMP1蛋白表达的阳性率显著低于对照组(P<0.05),COX-2及Stathmin蛋白表达的阳性率显著高于对照组(P<0.05)。实验组EMP1的m RNA水平显著低于对照组(P<0.05),COX-2及Stathmin m RNA水平显著高于对照组(P<0.05)。EMP1蛋白表达水平与上皮化生的严重程度呈负相关,COX-2及Stathmin蛋白表达水平与上皮化生的严重程度呈正相关。结论鼻腔鼻窦内翻性乳头状瘤中EMP1、COX-2及Stathmin蛋白的异常表达与上皮化生程度有关,联合检测EMP1、COX-2及Stathmin蛋白表达可评估鼻腔鼻窦内翻性乳头状瘤上皮异常重塑情况。展开更多
基金Supported by Hong Kong Research Grant Council,No.467109,467507the Scientif ic Research Fund of Zhejiang Provincial Ed-ucation Department,No.Y200906317+1 种基金the Wenzhou Science and Technology Bureau Program,No.Y20100017Qianjiang Talents Project of Zhejiang Province,No.2011R10058
文摘AIM:To investigate the role of transforming growth factor(TGF)-β-inducible early gene 1(TIEG1) in TGF-β-induced growth inhibition in hepatocellular carcinoma(HCC) cells.METHODS:Human hepatocyte and HCC cell lines with varied susceptibilities to TGF-β1 were tested by methylthiazoletetrazolium(MTT) assay.The expression changes of Smad2,Smad3,Smad4,Smad7,TIEG1 and TIEG2 gene following treatment with TGF-β1 in a TGF-β-sensitive hepatocyte cell line(MIHA),a TGF-β-sensitive hepatoma cell line(Hep3B) and two TGF-β-insensitive hepatoma cell lines(HepG2 and Bel7404) were examined.SiRNA targeting TIEG1 was transfected into Hep3B cells and the sensitivity of cells to TGF-β1 was examined.Overexpression of TIEG1 was induced by lentiviral-mediated transduction in TGF-β1-resistant hepatoma cell lines(Bel7404 and HepG2).MTT assay and 4',6-Diamidino-2-phenylindole staining were used to identify cell viability and apoptosis,respectively.The expression level of stathmin was measured by reverse transcriptase polymerase chain reaction and Western-blotting analysis,and stathmin promoter activity by TIEG1 was monitored by a luciferase reporter gene system.RESULTS:TIEG1 was significantly upregulated by TGF-β1 in the TGF-β1-sensitive HCC cell line,Hep3B,but not in the resistant cell lines.The suppression of TIEG1 by siRNAs decreased the sensitivity of Hep3B cells to TGF-β1,whereas the overexpression of TIEG1 mediated growth inhibition and apoptosis in TGF-β1-resistant HCC cell lines,which resembled those of TGF-β1-sensitive HCC cells treated with TGF-β1.Our data further suggested that stathmin was a direct target of TIEG1,as stathmin was signif icantly downregulated by TIEG1 overexpression,and stathmin promoter activity was inhibited by TIEG1 in a dose-dependent manner.CONCLUSION:Our data suggest that transactivation of TIEG1 conferred growth inhibition of TGF-β-susceptible human HCC cells.
文摘目的探讨以RNA干扰(RNAinterference,RNAi)技术靶向stathmin和mdr1基因逆转卵巢癌细胞紫杉醇耐药的可行性。方法分别构建靶向stathmin和mdr1基因的质粒:pGU6-GFP-neo-ST-MN1和pGU6-GFP-neo-MDR1;将质粒转染到卵巢癌紫杉醇耐药细胞株SKOV3/TAX。Real-ti me RT-PCR检测stathminm和mdr1的mRNA变化;Western blot检测其蛋白表达变化;荧光显微镜检测细胞凋亡情况;CCK-8法分析细胞对紫杉醇的敏感度。结果 Real-ti me RT-PCR及Western blot显示pGU6-GFP-neo-MDR1质粒对mdr1基因,pGU6-GFP-STMN1-294shRNA质粒对stathmin基因在mR-NA水平和蛋白水平均有明显抑制(P<0.05),荧光显微镜下共转染组细胞凋亡增多,CCK-8示共转染组逆转紫杉醇耐药效果最明显。结论体外RNAi可有效沉默卵巢癌紫杉醇耐药株SKOV3/TAX细胞内stathmin基因和mdr1基因,逆转其紫杉醇耐药。
文摘目的检测鼻腔鼻窦内翻性乳头状瘤中上皮细胞膜蛋白1(EMP1)、环氧化酶2(COX-2)及Stathmin蛋白的表达情况,分析其与上皮化生程度的相关性。方法选取2015年1月~2016年3月在本院就诊的鼻腔鼻窦内翻性乳头状瘤患者28例作为实验组,20例肥厚性鼻炎患者的下鼻甲黏膜组织作为对照组,采用免疫组化技术和q RT-PCR检测鼻腔鼻窦内翻性乳头状瘤、下鼻甲黏膜组织EMP1、COX-2以及Stathmin的表达情况,并比较三种因子在不同组织中的表达情况。结果实验组EMP1蛋白表达的阳性率显著低于对照组(P<0.05),COX-2及Stathmin蛋白表达的阳性率显著高于对照组(P<0.05)。实验组EMP1的m RNA水平显著低于对照组(P<0.05),COX-2及Stathmin m RNA水平显著高于对照组(P<0.05)。EMP1蛋白表达水平与上皮化生的严重程度呈负相关,COX-2及Stathmin蛋白表达水平与上皮化生的严重程度呈正相关。结论鼻腔鼻窦内翻性乳头状瘤中EMP1、COX-2及Stathmin蛋白的异常表达与上皮化生程度有关,联合检测EMP1、COX-2及Stathmin蛋白表达可评估鼻腔鼻窦内翻性乳头状瘤上皮异常重塑情况。
