AIM： To investigate the effects of Gli-1 small interference RNA （siRNA） on Huh7 cells, and the change of Bcl-2 expression in Huh7 cells. METHODS： Human hepatocellular carcinoma cells Huh7 were used. Cell viability...AIM： To investigate the effects of Gli-1 small interference RNA （siRNA） on Huh7 cells, and the change of Bcl-2 expression in Huh7 cells. METHODS： Human hepatocellular carcinoma cells Huh7 were used. Cell viability was analyzed by 3-（4, 5-Dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide （MTT） assay. The expressions of Gli-1 and Bcl-2 family members were detected by RT-PCR and Western blot. Apoptosis was detected by Flow cytometry using propidium iodide, measured by Hoechst 33258 staining using Advanced Fluorescence Microscopy and caspase-3 enzymatic assay. Cell growth was analyzed after treatment with Gli-1 siRNA and 5-fluorouracil （5-Fu）. RESULTS： Inhibition of Gli-1 mRNA in Huh7 cells through Gli-1 siRNA reduced cell viability. Gli-1 siRNA treatment also induced apoptosis by three criteria, increase in the sub-G1 cell cycle fraction, nuclear condensation, a morphologic change typical of apoptosis, and activation of caspase-3. Gli-1 siRNA was also able to down-regulate Bcl-2. However, Gli-1 siRNA resulted in no significant changes in Bcl-xl, Bax, Bad, and Bid. Furthermore, Gli-1 siRNA increased the cytotoxic effect of 5-Fu on Huh7 cell. CONCLUSION： Down-regulation of Bcl-2 plays an important role in apoptosis induced by Gli-1 siRNA in HCC cells. Combination Gli-1 siRNA with chemotherapeutic drug could represent a more promising strategy against HCC. The effects of the strategies need further investigation in vivo and may have potential clinical application.展开更多
Genetic diversity at Gli_1, Gli_2 and Glu_1 loci was investigated in 89 Sichuan wheat (Triticum aestivum L.) landraces by using acid polyacrylamide gel electrophoresis (APAGE) and SDS_PAGE. In these landraces, a total...Genetic diversity at Gli_1, Gli_2 and Glu_1 loci was investigated in 89 Sichuan wheat (Triticum aestivum L.) landraces by using acid polyacrylamide gel electrophoresis (APAGE) and SDS_PAGE. In these landraces, a total of 32 gliadin and 3 high_molecular_weight (HMW) glutenin patterns were observed. In total, 14, 15 and 5 alleles were identified at Gli_1, Gli_2 and Glu_1, respectively. At each locus, the alleles in higher frequency were Gli_A1a (89%), Gli_B1h (46%), Gli_D1a (65%), Gli_A2a (64%), Gli_B2j (45%), Gli_D2a (48%), Glu_A1c (99%), Glu_B1b (99%) and Glu_D1a (100%). The Nei's genetic variation index (H) of Sichuan wheat landraces was 0.370?6, varying from 0 to 0.708?7. The highest genetic diversity was found at Gli_B2 locus, while the lowest was found at Glu_D1. The genetic diversity at Gli loci was higher than that of Glu_1 loci among these landraces, but it was much lower than that of modern wheat cultivars. These results indicated a narrow genetic base of Sichuan wheat landraces. In this study, “Chengdu_guangtou” had the identical gliadin and HMW_glutenin patterns with “Chinese Spring”, further supporting the proposal that “Chinese Spring” is a strain of “Chengdu_guangtou”.展开更多
文摘目的:探讨胶质瘤相关癌基因同源蛋白-1(Gli-1)及上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白Snail、E-cadherin及N-cadherin在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达及临床意义。方法:采用免疫组织化学法检测天津市胸科医院2010年1月至2012年2月经手术切除送检的67例NSCLC组织及20例正常肺组织中Gli-1、Snail、E-cadherin及N-cadherin蛋白的表达情况,分析其与患者临床病理特征及预后的关系;采用逆转录多聚酶链反应(RT-PCR)法测定Gli-1、Snail、E-cadherin及N-cadherin基因在20例NSCLC及其对应癌旁正常肺组织中m RNA的表达情况。结果:1)Gli-1、Snail、E-cadherin及N-cadherin在NSCLC组织中的阳性表达率分别为61.19%(41/67)、50.75%(34/67)、56.72%(38/67)、53.73%(36/67),在正常肺组织中的阳性表达率为20%(4/20)、10%(2/20)、100%(20/20)、5%(1/20),表达差异具有统计学意义(P<0.05);2)患者癌组织中Gli-1高表达与淋巴结转移、分化程度和临床分期密切相关(P<0.05),与性别、年龄、肿物大小和病理分型无相关性;Gli-1的表达与E-cadherin的表达呈负相关(r=-0.325,P<0.05),与Snail、N-cadherin的表达呈正相关(r=0.379、r=0.490,P<0.05);3)RT-PCR结果显示,Gli-1、Snail和N-cadherin m RNA的表达量高于正常余肺组织(P<0.05),E-cadherin m RNA的表达量低于正常余肺组织(P<0.05);4)Gli-1、Snail和N-cadherin高表达的患者表现出更差的预后和更低的生存率(P<0.05),E-cadherin阴性组的患者较阳性组患者显示出更差的预后和更低的生存率,差异具有统计学意义(P<0.05),Cox比例风险回归模型分析结果显示,E-cadherin阴性、淋巴结转移、临床分期、分化程度是影响NSCLC患者预后的独立危险因素。结论:在NSCLC中,Hedgehog信号通路的异常活化与EMT存在密切相关性,Gli-1与EMT相关蛋白的表达对判断NSCLC患者的临床特征及预后具有一定提示作用。
基金The National Science Foundation of China, No. 30672053
文摘AIM： To investigate the effects of Gli-1 small interference RNA （siRNA） on Huh7 cells, and the change of Bcl-2 expression in Huh7 cells. METHODS： Human hepatocellular carcinoma cells Huh7 were used. Cell viability was analyzed by 3-（4, 5-Dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide （MTT） assay. The expressions of Gli-1 and Bcl-2 family members were detected by RT-PCR and Western blot. Apoptosis was detected by Flow cytometry using propidium iodide, measured by Hoechst 33258 staining using Advanced Fluorescence Microscopy and caspase-3 enzymatic assay. Cell growth was analyzed after treatment with Gli-1 siRNA and 5-fluorouracil （5-Fu）. RESULTS： Inhibition of Gli-1 mRNA in Huh7 cells through Gli-1 siRNA reduced cell viability. Gli-1 siRNA treatment also induced apoptosis by three criteria, increase in the sub-G1 cell cycle fraction, nuclear condensation, a morphologic change typical of apoptosis, and activation of caspase-3. Gli-1 siRNA was also able to down-regulate Bcl-2. However, Gli-1 siRNA resulted in no significant changes in Bcl-xl, Bax, Bad, and Bid. Furthermore, Gli-1 siRNA increased the cytotoxic effect of 5-Fu on Huh7 cell. CONCLUSION： Down-regulation of Bcl-2 plays an important role in apoptosis induced by Gli-1 siRNA in HCC cells. Combination Gli-1 siRNA with chemotherapeutic drug could represent a more promising strategy against HCC. The effects of the strategies need further investigation in vivo and may have potential clinical application.
文摘Genetic diversity at Gli_1, Gli_2 and Glu_1 loci was investigated in 89 Sichuan wheat (Triticum aestivum L.) landraces by using acid polyacrylamide gel electrophoresis (APAGE) and SDS_PAGE. In these landraces, a total of 32 gliadin and 3 high_molecular_weight (HMW) glutenin patterns were observed. In total, 14, 15 and 5 alleles were identified at Gli_1, Gli_2 and Glu_1, respectively. At each locus, the alleles in higher frequency were Gli_A1a (89%), Gli_B1h (46%), Gli_D1a (65%), Gli_A2a (64%), Gli_B2j (45%), Gli_D2a (48%), Glu_A1c (99%), Glu_B1b (99%) and Glu_D1a (100%). The Nei's genetic variation index (H) of Sichuan wheat landraces was 0.370?6, varying from 0 to 0.708?7. The highest genetic diversity was found at Gli_B2 locus, while the lowest was found at Glu_D1. The genetic diversity at Gli loci was higher than that of Glu_1 loci among these landraces, but it was much lower than that of modern wheat cultivars. These results indicated a narrow genetic base of Sichuan wheat landraces. In this study, “Chengdu_guangtou” had the identical gliadin and HMW_glutenin patterns with “Chinese Spring”, further supporting the proposal that “Chinese Spring” is a strain of “Chengdu_guangtou”.
