AIM:To investigate the regulative effect of miRNA-3383p(miR-338-3p) on cell growth in colorectal carcinoma(CRC).METHODS:The lentiviral vector pLV-THM-miR-338-3p and pLV-THM-miR-338-3p-inhibitor were constructed.The re...AIM:To investigate the regulative effect of miRNA-3383p(miR-338-3p) on cell growth in colorectal carcinoma(CRC).METHODS:The lentiviral vector pLV-THM-miR-338-3p and pLV-THM-miR-338-3p-inhibitor were constructed.The recombinant viral vector encoding the pre-miR338-3p or miR-338-3p-inhibitor and the two packaging plasmids psPAX2 and pMD2.G were cotransfected into human embryonic kidney 293T cells to package lentivirus.The supernatant containing the lentivirus particles was harvested to determine the viral titer,and this supernatant was then used to transduce CRCderived cell line,SW-620.Flow cytometry was utilized for sorting the green fluorescent protein(GFP) + cells to establish the SW-620 cell line stably expressing premiR-338-3p or miR-338-3p-inhibitor.Moreover,the expression of miR-338-3p was determined by real-time reverse transcriptase polymerase chain reaction,andWestern blotting was used to detect the expression of the smoothened(SMO,the possible target of miR-3383p) protein in SW-620 cells.Furthermore,the status of CRC cell proliferation and apoptosis were detected by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay and flow cytometry,respectively.RESULTS:Restriction enzyme digestion and DNA sequencing demonstrated that the lentiviral vector pLVTHM-miR-338-3p and pLV-THM-miR-338-3p-inhibitor were constructed successfully.GFP was expressed after the SW-620 cells were transduced by the lentivirus.Expression of miR-338-3p in SW-620 cells transduced with the lentivirus pLV-THM-miR-338-3p was significantly increased(relative expression 3.91 ± 0.51 vs 2.36 ± 0.44,P < 0.01).Furthermore,overexpression of miR-338-3p inhibited the expression of SMO protein in SW-620 cells,which showed obviously suppressed proliferation ability [cellular proliferation inhibition rate(CPIR) 61.9% ± 5.2% vs 41.6% ± 4.8%,P < 0.01].Expression of miR-338-3p in SW-620 cells transduced with the lentivirus pLV-THM-miR-338-3p-inhibitor was significantly decreased(relative expression 0.92 ± 0.29 vs 2.36 ± 0.44,P < 0.01).Moreover,the downregulated expression of miR-338-3p caused upregulated expression of the SMO protein in SW-620 cells,which showed significantly enhanced proliferation ability(CPIR 19.2% ± 3.8% vs 41.6% ± 4.8%,P < 0.01).However,anti-SMO-siRNA largely,but not completely,reversed the effects induced by blockage of miR-338-3p,suggesting that the regulative effect of miR-338-3p on CRC cell growth was indeed mediated by SMO.CONCLUSION:miR-338-3p could suppress CRC growth by inhibiting SMO protein expression.展开更多
目的探讨通过沉默Smoothened(Smo)基因抑制Hedgehog通路后,小鼠胚胎成纤维细胞NIH/3T3中mRNA表达谱的变化。方法提取Smo沉默和对照NIH/3T3细胞的总RNA,并进行测序。与对照组比较,分析差异表达的mRNA,通过BLAST2GO和京都基因与基因组百...目的探讨通过沉默Smoothened(Smo)基因抑制Hedgehog通路后,小鼠胚胎成纤维细胞NIH/3T3中mRNA表达谱的变化。方法提取Smo沉默和对照NIH/3T3细胞的总RNA,并进行测序。与对照组比较,分析差异表达的mRNA,通过BLAST2GO和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析和预测这些差异表达mRNA的相关功能。选取差异表达mRNA进行实时定量PCR验证。结果通过测序在Smo沉默的NIH/3T3细胞中发现有2289个mRNA差异表达,其中1646个mRNA在Smo沉默后表达下调,而643个mRNA表达上调。荧光实时定量PCR结果与测序结果一致。生物信息学分析显示,差异表达mRNA参与细胞过程、单有机体过程、生物学调节。差异表达mRNA富集在轴突导向(ko04360)、癌症通路(ko05200)、癌症中的microRNA(ko05206)等通路。利用GEPIA数据库分析CXCL12、IL-33、TGF-β2和FGF10及其受体在食管癌和正常食管组织中的表达情况,和预后的预测分析。结论在NIH/3T3细胞中沉默Smo抑制Hedgehog通路可能会通过细胞因子CXCL12、IL-33、TGF-β2和FGF10影响肿瘤细胞的增殖。展开更多
The aim of this study was to develop an improved thin sea ice thickness(SIT)retrieval algorithm in the Arctic Ocean from the Soil Moisture Ocean Salinity and Soil Moisture Active Passive L-band radiometer data.This SI...The aim of this study was to develop an improved thin sea ice thickness(SIT)retrieval algorithm in the Arctic Ocean from the Soil Moisture Ocean Salinity and Soil Moisture Active Passive L-band radiometer data.This SIT retrieval algorithm was trained using the simulated SIT from the cumulative freezing degree days model during the freeze-up period over five carefully selected regions in the Beaufort,Chukchi,East Siberian,Laptev and Kara seas and utilized the microwave polarization ratio(PR)at incidence angle of 40°.The improvements of the proposed retrieval algorithm include the correction for the sea ice concentration impact,reliable reference SIT data over different representative regions of the Arctic Ocean and the utilization of microwave polarization ratio that is independent of ice temperature.The relationship between the SIT and PR was found to be almost stable across the five selected regions.The SIT retrievals were then compared to other two existing algorithms(i.e.,UH_SIT from the University of Hamburg and UB_SIT from the University of Bremen)and validated against independent SIT data obtained from moored upward looking sonars(ULS)and airborne electromagnetic(EM)induction sensors.The results suggest that the proposed algorithm could achieve comparable accuracies to UH_SIT and UB_SIT with root mean square error(RMSE)being about 0.20 m when validating using ULS SIT data and outperformed the UH_SIT and UB_SIT with RMSE being about 0.21 m when validatng using EM SIT data.The proposed algorithm can be used for thin sea ice thickness(<1.0 m)estimation in the Arctic Ocean and requires less auxiliary data in the SIT retrieval procedure which makes its implementation more practical.展开更多
基金Supported by National Natural Science Foundation of China,No.81101896
文摘AIM:To investigate the regulative effect of miRNA-3383p(miR-338-3p) on cell growth in colorectal carcinoma(CRC).METHODS:The lentiviral vector pLV-THM-miR-338-3p and pLV-THM-miR-338-3p-inhibitor were constructed.The recombinant viral vector encoding the pre-miR338-3p or miR-338-3p-inhibitor and the two packaging plasmids psPAX2 and pMD2.G were cotransfected into human embryonic kidney 293T cells to package lentivirus.The supernatant containing the lentivirus particles was harvested to determine the viral titer,and this supernatant was then used to transduce CRCderived cell line,SW-620.Flow cytometry was utilized for sorting the green fluorescent protein(GFP) + cells to establish the SW-620 cell line stably expressing premiR-338-3p or miR-338-3p-inhibitor.Moreover,the expression of miR-338-3p was determined by real-time reverse transcriptase polymerase chain reaction,andWestern blotting was used to detect the expression of the smoothened(SMO,the possible target of miR-3383p) protein in SW-620 cells.Furthermore,the status of CRC cell proliferation and apoptosis were detected by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay and flow cytometry,respectively.RESULTS:Restriction enzyme digestion and DNA sequencing demonstrated that the lentiviral vector pLVTHM-miR-338-3p and pLV-THM-miR-338-3p-inhibitor were constructed successfully.GFP was expressed after the SW-620 cells were transduced by the lentivirus.Expression of miR-338-3p in SW-620 cells transduced with the lentivirus pLV-THM-miR-338-3p was significantly increased(relative expression 3.91 ± 0.51 vs 2.36 ± 0.44,P < 0.01).Furthermore,overexpression of miR-338-3p inhibited the expression of SMO protein in SW-620 cells,which showed obviously suppressed proliferation ability [cellular proliferation inhibition rate(CPIR) 61.9% ± 5.2% vs 41.6% ± 4.8%,P < 0.01].Expression of miR-338-3p in SW-620 cells transduced with the lentivirus pLV-THM-miR-338-3p-inhibitor was significantly decreased(relative expression 0.92 ± 0.29 vs 2.36 ± 0.44,P < 0.01).Moreover,the downregulated expression of miR-338-3p caused upregulated expression of the SMO protein in SW-620 cells,which showed significantly enhanced proliferation ability(CPIR 19.2% ± 3.8% vs 41.6% ± 4.8%,P < 0.01).However,anti-SMO-siRNA largely,but not completely,reversed the effects induced by blockage of miR-338-3p,suggesting that the regulative effect of miR-338-3p on CRC cell growth was indeed mediated by SMO.CONCLUSION:miR-338-3p could suppress CRC growth by inhibiting SMO protein expression.
文摘目的探讨通过沉默Smoothened(Smo)基因抑制Hedgehog通路后,小鼠胚胎成纤维细胞NIH/3T3中mRNA表达谱的变化。方法提取Smo沉默和对照NIH/3T3细胞的总RNA,并进行测序。与对照组比较,分析差异表达的mRNA,通过BLAST2GO和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析和预测这些差异表达mRNA的相关功能。选取差异表达mRNA进行实时定量PCR验证。结果通过测序在Smo沉默的NIH/3T3细胞中发现有2289个mRNA差异表达,其中1646个mRNA在Smo沉默后表达下调,而643个mRNA表达上调。荧光实时定量PCR结果与测序结果一致。生物信息学分析显示,差异表达mRNA参与细胞过程、单有机体过程、生物学调节。差异表达mRNA富集在轴突导向(ko04360)、癌症通路(ko05200)、癌症中的microRNA(ko05206)等通路。利用GEPIA数据库分析CXCL12、IL-33、TGF-β2和FGF10及其受体在食管癌和正常食管组织中的表达情况,和预后的预测分析。结论在NIH/3T3细胞中沉默Smo抑制Hedgehog通路可能会通过细胞因子CXCL12、IL-33、TGF-β2和FGF10影响肿瘤细胞的增殖。
基金The National Natural Science Foundation of China under contract Nos 41830536 and 41925027the Guangdong Natural Science Foundation under contract No.2023A1515011235the Innovation Group Project of Southern Marine Science and Engineering Guangdong Laboratory(Zhuhai)under contract No.311021008.
文摘The aim of this study was to develop an improved thin sea ice thickness(SIT)retrieval algorithm in the Arctic Ocean from the Soil Moisture Ocean Salinity and Soil Moisture Active Passive L-band radiometer data.This SIT retrieval algorithm was trained using the simulated SIT from the cumulative freezing degree days model during the freeze-up period over five carefully selected regions in the Beaufort,Chukchi,East Siberian,Laptev and Kara seas and utilized the microwave polarization ratio(PR)at incidence angle of 40°.The improvements of the proposed retrieval algorithm include the correction for the sea ice concentration impact,reliable reference SIT data over different representative regions of the Arctic Ocean and the utilization of microwave polarization ratio that is independent of ice temperature.The relationship between the SIT and PR was found to be almost stable across the five selected regions.The SIT retrievals were then compared to other two existing algorithms(i.e.,UH_SIT from the University of Hamburg and UB_SIT from the University of Bremen)and validated against independent SIT data obtained from moored upward looking sonars(ULS)and airborne electromagnetic(EM)induction sensors.The results suggest that the proposed algorithm could achieve comparable accuracies to UH_SIT and UB_SIT with root mean square error(RMSE)being about 0.20 m when validating using ULS SIT data and outperformed the UH_SIT and UB_SIT with RMSE being about 0.21 m when validatng using EM SIT data.The proposed algorithm can be used for thin sea ice thickness(<1.0 m)estimation in the Arctic Ocean and requires less auxiliary data in the SIT retrieval procedure which makes its implementation more practical.
