目的:探讨microRNA-29c对人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2)生物学特性的影响。方法:培养1种人正常胰腺上皮细胞(HPDE)及4种人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2),采用real-time PCR法观察5种细胞系...目的:探讨microRNA-29c对人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2)生物学特性的影响。方法:培养1种人正常胰腺上皮细胞(HPDE)及4种人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2),采用real-time PCR法观察5种细胞系中microRNA-29c的表达差异,以microRNA-29c过表达腺病毒感染的PANC-1和MIA Pa Ca2细胞作为实验组,以空载体感染的PANC-1和MIA Pa Ca2细胞作为阴性对照组,采用细胞划痕实验、Transwell法检测两组细胞体外侵袭能力,Western blot检测两组细胞上皮间充质转化(EMT)相关蛋白波形蛋白(Vimentin)及E-钙粘蛋白(E-cadherin)的表达。结果:real-time-PCR显示各胰腺癌细胞系中microRNA-29c水平明显低于正常胰腺细胞系(P<0.05),细胞划痕实验发现感染腺病毒48 h后实验组PANC-1、MIA Pa Ca-2细胞的迁移距离明显短于阴性对照组(P<0.05),Transwell小室细胞侵袭实验发现实验组PANC-1和MIA Pa Ca-2细胞侵袭数量明显低于阴性对照组(P<0.05);Western blot蛋白免疫印迹结果显示PANC-1和MIA Pa Ca-2细胞过表达microRNA-29c后,Vimentin表达减少,E-cadherin表达增加。结论:microRNA-29c的过表达可有效抑制胰腺癌细胞的体外侵袭与转移,可能与Vimentin表达减少,E-cadherin表达增加有关,有望成为胰腺癌生物治疗的潜在靶点。展开更多
AIM： To determine the role of microRNA （miRNA）-29a and miRNA-29c in the regulation of apoptosis in early rat diabetic cataract formation.METHODS： Streptozotocin （STZ）-induced diabetic Sprague-Dawley （SD） rats ...AIM： To determine the role of microRNA （miRNA）-29a and miRNA-29c in the regulation of apoptosis in early rat diabetic cataract formation.METHODS： Streptozotocin （STZ）-induced diabetic Sprague-Dawley （SD） rats were used in the study. The expression level of miRNA-29a, miRNA-29c, and BCL2-modifying factor （BMF） in lens epithelial cells （LECs） samples were measured using quantitative real-time polymerase chain reaction. Prediction algorithms of miRanda, TargetScan 6.2, and mirRDB to perform a miRNA gene network analysis were used for the potential miRNA-29a and miRNA-29c targets. RESULTS： The miRNA-29a and miRNA-29c expression levels were all significantly lower in the control group compared to the 2 and 4wk diabetic samples （P〈0.01）. The network analysis indicated that one miRNA-29a and miRNA-29c targets was BMF. There was significantly higher expression of BMF mRNA compared to the normal controls （P〈0.01）. CONCLUSION： Apoptosis occurs in rat LECs following high blood glucose exposure. It is likely that apoptosis during diabetic cataract formation involves the decreased expression of miRNA-29a and miRNA-29c and the increased expression of BMF.展开更多
文摘目的:探讨microRNA-29c对人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2)生物学特性的影响。方法:培养1种人正常胰腺上皮细胞(HPDE)及4种人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2),采用real-time PCR法观察5种细胞系中microRNA-29c的表达差异,以microRNA-29c过表达腺病毒感染的PANC-1和MIA Pa Ca2细胞作为实验组,以空载体感染的PANC-1和MIA Pa Ca2细胞作为阴性对照组,采用细胞划痕实验、Transwell法检测两组细胞体外侵袭能力,Western blot检测两组细胞上皮间充质转化(EMT)相关蛋白波形蛋白(Vimentin)及E-钙粘蛋白(E-cadherin)的表达。结果:real-time-PCR显示各胰腺癌细胞系中microRNA-29c水平明显低于正常胰腺细胞系(P<0.05),细胞划痕实验发现感染腺病毒48 h后实验组PANC-1、MIA Pa Ca-2细胞的迁移距离明显短于阴性对照组(P<0.05),Transwell小室细胞侵袭实验发现实验组PANC-1和MIA Pa Ca-2细胞侵袭数量明显低于阴性对照组(P<0.05);Western blot蛋白免疫印迹结果显示PANC-1和MIA Pa Ca-2细胞过表达microRNA-29c后,Vimentin表达减少,E-cadherin表达增加。结论:microRNA-29c的过表达可有效抑制胰腺癌细胞的体外侵袭与转移,可能与Vimentin表达减少,E-cadherin表达增加有关,有望成为胰腺癌生物治疗的潜在靶点。
文摘AIM： To determine the role of microRNA （miRNA）-29a and miRNA-29c in the regulation of apoptosis in early rat diabetic cataract formation.METHODS： Streptozotocin （STZ）-induced diabetic Sprague-Dawley （SD） rats were used in the study. The expression level of miRNA-29a, miRNA-29c, and BCL2-modifying factor （BMF） in lens epithelial cells （LECs） samples were measured using quantitative real-time polymerase chain reaction. Prediction algorithms of miRanda, TargetScan 6.2, and mirRDB to perform a miRNA gene network analysis were used for the potential miRNA-29a and miRNA-29c targets. RESULTS： The miRNA-29a and miRNA-29c expression levels were all significantly lower in the control group compared to the 2 and 4wk diabetic samples （P〈0.01）. The network analysis indicated that one miRNA-29a and miRNA-29c targets was BMF. There was significantly higher expression of BMF mRNA compared to the normal controls （P〈0.01）. CONCLUSION： Apoptosis occurs in rat LECs following high blood glucose exposure. It is likely that apoptosis during diabetic cataract formation involves the decreased expression of miRNA-29a and miRNA-29c and the increased expression of BMF.
