目的探讨mi R-26a在乳腺癌组织和细胞中表达量的改变及其对人乳腺癌细胞增殖和侵袭的影响。方法采用实时荧光定量PCR(Real-time PCR)法检测20例患者乳腺癌组织及对应的癌旁组织、人乳腺癌细胞MCF-7、BT474及健康者乳腺细胞MCF-10A中mi R...目的探讨mi R-26a在乳腺癌组织和细胞中表达量的改变及其对人乳腺癌细胞增殖和侵袭的影响。方法采用实时荧光定量PCR(Real-time PCR)法检测20例患者乳腺癌组织及对应的癌旁组织、人乳腺癌细胞MCF-7、BT474及健康者乳腺细胞MCF-10A中mi R-26a的表达,用Western bolt法检测COX-2的表达。MCF-7及BT474细胞分别转染mi R-NC和mi R-26a,采用Western blot法检测转染后细胞中COX-2的表达水平,同时采用CCK-8和克隆形成实验检测转染后细胞的增殖和侵袭情况。结果mi R-26a在乳腺癌组织中的表达明显低于癌旁组织(t=20.33,P=0.001),在MCF-7和BT474细胞中的表达明显低于MCF-10A细胞(Dunnett t test I-J=-0.031,P=0.001)。COX-2在乳腺癌组织和细胞中的表达明显高于癌旁组织(t=18.01,P=0.002)和健康者乳腺细胞(Dunnett t test I-J=-0.028,P=0.000)。转染mi R-26a后,乳腺癌细胞内COX-2的表达量显著下调。CCK-8和克隆形成实验结果显示,过表达mi R-26a能明显抑制乳腺癌细胞的增殖(F=6.032,P=0.013)和侵袭(Dunnett t test I-J=-0.21,P=0.037)。结论过表达mi R-26a可以通过下调乳腺癌细胞中COX-2的表达,抑制乳腺癌细胞的增殖和侵袭。展开更多
Objective:To observe the expression of Long non-coding RNA antisense mitochondrial non-coding RNA-2 (ASncmtRNA-2) in high glucose (HG) treated human renal mesangial cells (HRMCs) and the role of ASncmtRNA-2 in oxidati...Objective:To observe the expression of Long non-coding RNA antisense mitochondrial non-coding RNA-2 (ASncmtRNA-2) in high glucose (HG) treated human renal mesangial cells (HRMCs) and the role of ASncmtRNA-2 in oxidative stress mediated diabetic nephropathy (DN) fibrosis.Methods: The expression levels of ASncmtRNA-2、transforming growth factorβ1 (TGF-β1) and fibronectin (FN) mRNA in cultured HRMCs were measured by qRT-PCR. In addition, relative reactive oxygen species (ROS) levels in HRMCs were detected with the non-fluorescent probe DCFH-DA assays.Results: Compared with 0h, the expression of ASncmtRNA-2 remained unchanged in all groups at 8 h post treatment. However, the level of ASncmtRNA-2 mRNA was increased significantly in HG and HG+NG-nitro-L-Arginine methylester (L-NAME) treated cells compared with low glucose (LG) treated cells from 16h onwards, while the level of ASncmtRNA-2 mRNA in the HG+L-NAME group was decreased compared with the HG group. Moreover, ROS fluorescence was significantly up-regulated in HG-treated cells compared with LG-treated cells, while the ROS fluorescence in HG+L-NAME group was suppressed compared with HG-treated cells. In addition, Levels of ASncmtRNA-2, TGF-β1 and FN mRNA were significantly up-regulated in HG treated cells compared with LG treated cells while Levels of ASncmtRNA-2, TGF-β1 and FN mRNA in HG+L-NAME group were down-regulated compared with HG group. Finally, the expression of ASncmtRNA-2, TGF-β1 and FN mRNA were significantly decreased in HG+ASncmtRNA-2 siRNA group compared with HG group.Conclusion: ASncmtRNA-2 was up-regulated in HG treated cells and may promote glomerular fibrosis in DN via positively regulating the expression of pro-fibrotic factors. These findings may provide novel potential therapeutic treatments for DN.展开更多
文摘目的探讨mi R-26a在乳腺癌组织和细胞中表达量的改变及其对人乳腺癌细胞增殖和侵袭的影响。方法采用实时荧光定量PCR(Real-time PCR)法检测20例患者乳腺癌组织及对应的癌旁组织、人乳腺癌细胞MCF-7、BT474及健康者乳腺细胞MCF-10A中mi R-26a的表达,用Western bolt法检测COX-2的表达。MCF-7及BT474细胞分别转染mi R-NC和mi R-26a,采用Western blot法检测转染后细胞中COX-2的表达水平,同时采用CCK-8和克隆形成实验检测转染后细胞的增殖和侵袭情况。结果mi R-26a在乳腺癌组织中的表达明显低于癌旁组织(t=20.33,P=0.001),在MCF-7和BT474细胞中的表达明显低于MCF-10A细胞(Dunnett t test I-J=-0.031,P=0.001)。COX-2在乳腺癌组织和细胞中的表达明显高于癌旁组织(t=18.01,P=0.002)和健康者乳腺细胞(Dunnett t test I-J=-0.028,P=0.000)。转染mi R-26a后,乳腺癌细胞内COX-2的表达量显著下调。CCK-8和克隆形成实验结果显示,过表达mi R-26a能明显抑制乳腺癌细胞的增殖(F=6.032,P=0.013)和侵袭(Dunnett t test I-J=-0.21,P=0.037)。结论过表达mi R-26a可以通过下调乳腺癌细胞中COX-2的表达,抑制乳腺癌细胞的增殖和侵袭。
文摘Objective:To observe the expression of Long non-coding RNA antisense mitochondrial non-coding RNA-2 (ASncmtRNA-2) in high glucose (HG) treated human renal mesangial cells (HRMCs) and the role of ASncmtRNA-2 in oxidative stress mediated diabetic nephropathy (DN) fibrosis.Methods: The expression levels of ASncmtRNA-2、transforming growth factorβ1 (TGF-β1) and fibronectin (FN) mRNA in cultured HRMCs were measured by qRT-PCR. In addition, relative reactive oxygen species (ROS) levels in HRMCs were detected with the non-fluorescent probe DCFH-DA assays.Results: Compared with 0h, the expression of ASncmtRNA-2 remained unchanged in all groups at 8 h post treatment. However, the level of ASncmtRNA-2 mRNA was increased significantly in HG and HG+NG-nitro-L-Arginine methylester (L-NAME) treated cells compared with low glucose (LG) treated cells from 16h onwards, while the level of ASncmtRNA-2 mRNA in the HG+L-NAME group was decreased compared with the HG group. Moreover, ROS fluorescence was significantly up-regulated in HG-treated cells compared with LG-treated cells, while the ROS fluorescence in HG+L-NAME group was suppressed compared with HG-treated cells. In addition, Levels of ASncmtRNA-2, TGF-β1 and FN mRNA were significantly up-regulated in HG treated cells compared with LG treated cells while Levels of ASncmtRNA-2, TGF-β1 and FN mRNA in HG+L-NAME group were down-regulated compared with HG group. Finally, the expression of ASncmtRNA-2, TGF-β1 and FN mRNA were significantly decreased in HG+ASncmtRNA-2 siRNA group compared with HG group.Conclusion: ASncmtRNA-2 was up-regulated in HG treated cells and may promote glomerular fibrosis in DN via positively regulating the expression of pro-fibrotic factors. These findings may provide novel potential therapeutic treatments for DN.
