[目的]探索SLC1A3在前列腺癌中的表达以及对前列腺癌DU145细胞恶性转移的影响。[方法]蛋白免疫印迹分析前列腺癌细胞与前列腺正常细胞中SLC1A3的表达;将人前列腺癌DU145细胞分为3个实验组:siRNA NC组、siRNA SLC1A3组与LY294002组。成...[目的]探索SLC1A3在前列腺癌中的表达以及对前列腺癌DU145细胞恶性转移的影响。[方法]蛋白免疫印迹分析前列腺癌细胞与前列腺正常细胞中SLC1A3的表达;将人前列腺癌DU145细胞分为3个实验组:siRNA NC组、siRNA SLC1A3组与LY294002组。成球实验法检测前列腺癌DU145细胞的生长活性;细胞划痕实验分析前列腺癌DU145细胞的迁移能力;蛋白免疫印迹实验检测DU145细胞中PI3K/AKT蛋白的表达;TUNEL实验检测前列腺癌DU145细胞的凋亡率。[结果]与前列腺正常细胞相比,前列腺癌细胞中SLC1A3的表达上调(0.31±0.05 vs 0.82±0.07,P<0.05)。与siRNA NC组相比,siRNA SLC1A3和LY294002组的前列腺癌DU145细胞生长活性减弱(P<0.05);siRNA SLC1A3和LY294002组的前列腺癌DU145细胞迁移数量下降(P<0.05);siRNA SLC1A3和LY294002组的前列腺癌DU145细胞凋亡率增加(P<0.05);siRNA SLC1A3组的前列腺癌DU145细胞的PI3K/AKT蛋白表达降低(0.79±0.09 vs 0.29±0.03 vs 0.26±0.05;0.75±0.21 vs 0.22±0.05 vs 0.19±0.07,P<0.05)。[结论]SLC1A3在前列腺癌细胞中表达上调,抑制SLC1A3表达后,前列腺癌DU145细胞的生长活性以及迁移能力降低,凋亡率增加,这一过程与SLC1A3调节PI3K/AKT途径蛋白的表达相关。展开更多
AIM:To identify the genetic factors underlying anophthalmia and microphthalmia(A/M),and to perform computational analysis to verify the pathophysiological mechanisms of the disease.METHODS:This study investigated a co...AIM:To identify the genetic factors underlying anophthalmia and microphthalmia(A/M),and to perform computational analysis to verify the pathophysiological mechanisms of the disease.METHODS:This study investigated a consanguineous Pakistani family with multiple affected individuals.Clinical evaluations were conducted using A-Scan and ophthalmic B-Scan ultrasonography(B-Scan).To identify the diseasecausing variant,whole exome sequencing(WES)and Sanger sequencing were performed.In silico functional analyses were carried out using AlphaFold(for protein modeling)and ClusPro(for protein docking analysis)tools,and the hydrodynamic properties of the protein were determined via molecular dynamics simulations.RESULTS:Clinical analysis of the five patients revealed severe phenotypes of bilateral anophthalmia.Ocular B-Scan did not detect ocular tissue or intraocular fluid,thus confirming the diagnosis of anophthalmia in all patients.Due to these structural defects,all patients exhibited complete blindness and absence of light perception;additionally,two patients had mild to moderate intellectual disability.Genetic analysis identified a splice site variant[NM_000693.2:c.884-2_885dup;p.(Asp296SerfsTer35)]in the 9^(th)exon of the ALDH1A3 gene.CONCLUSION:The present study expands the genetic spectrum of ALDH1A3 and contributes to establishing the genotype-phenotype correlation for this gene.展开更多
文摘[目的]探索SLC1A3在前列腺癌中的表达以及对前列腺癌DU145细胞恶性转移的影响。[方法]蛋白免疫印迹分析前列腺癌细胞与前列腺正常细胞中SLC1A3的表达;将人前列腺癌DU145细胞分为3个实验组:siRNA NC组、siRNA SLC1A3组与LY294002组。成球实验法检测前列腺癌DU145细胞的生长活性;细胞划痕实验分析前列腺癌DU145细胞的迁移能力;蛋白免疫印迹实验检测DU145细胞中PI3K/AKT蛋白的表达;TUNEL实验检测前列腺癌DU145细胞的凋亡率。[结果]与前列腺正常细胞相比,前列腺癌细胞中SLC1A3的表达上调(0.31±0.05 vs 0.82±0.07,P<0.05)。与siRNA NC组相比,siRNA SLC1A3和LY294002组的前列腺癌DU145细胞生长活性减弱(P<0.05);siRNA SLC1A3和LY294002组的前列腺癌DU145细胞迁移数量下降(P<0.05);siRNA SLC1A3和LY294002组的前列腺癌DU145细胞凋亡率增加(P<0.05);siRNA SLC1A3组的前列腺癌DU145细胞的PI3K/AKT蛋白表达降低(0.79±0.09 vs 0.29±0.03 vs 0.26±0.05;0.75±0.21 vs 0.22±0.05 vs 0.19±0.07,P<0.05)。[结论]SLC1A3在前列腺癌细胞中表达上调,抑制SLC1A3表达后,前列腺癌DU145细胞的生长活性以及迁移能力降低,凋亡率增加,这一过程与SLC1A3调节PI3K/AKT途径蛋白的表达相关。
基金Supported by Taif University,Taif,Saudi Arabia(TU-DSPP-2024-05).
文摘AIM:To identify the genetic factors underlying anophthalmia and microphthalmia(A/M),and to perform computational analysis to verify the pathophysiological mechanisms of the disease.METHODS:This study investigated a consanguineous Pakistani family with multiple affected individuals.Clinical evaluations were conducted using A-Scan and ophthalmic B-Scan ultrasonography(B-Scan).To identify the diseasecausing variant,whole exome sequencing(WES)and Sanger sequencing were performed.In silico functional analyses were carried out using AlphaFold(for protein modeling)and ClusPro(for protein docking analysis)tools,and the hydrodynamic properties of the protein were determined via molecular dynamics simulations.RESULTS:Clinical analysis of the five patients revealed severe phenotypes of bilateral anophthalmia.Ocular B-Scan did not detect ocular tissue or intraocular fluid,thus confirming the diagnosis of anophthalmia in all patients.Due to these structural defects,all patients exhibited complete blindness and absence of light perception;additionally,two patients had mild to moderate intellectual disability.Genetic analysis identified a splice site variant[NM_000693.2:c.884-2_885dup;p.(Asp296SerfsTer35)]in the 9^(th)exon of the ALDH1A3 gene.CONCLUSION:The present study expands the genetic spectrum of ALDH1A3 and contributes to establishing the genotype-phenotype correlation for this gene.
