Background:Breast cancer(BC)is a common malignancy with highly female incidence.So far the function of notoginsenoside R1(NGR1),the extract from Panax notoginseng,has not been clearly elucidated in BC.Methods:Optimal ...Background:Breast cancer(BC)is a common malignancy with highly female incidence.So far the function of notoginsenoside R1(NGR1),the extract from Panax notoginseng,has not been clearly elucidated in BC.Methods:Optimal culture concentration and time of NGR1 were investigated by cell counting kit-8 assay.Cell proliferation ability was measured by colony formation assays.Transwell assay was used to detect the effect of NGR1 on cell migration and invasion.The apoptosis rate of cells between each group was measured by TUNEL assay.Results:NGR1 treatment has an inhibitory efect on proliferation,migration,invasion,and angiogenesis and a stimulating effect on cell cycle arrest and apoptosis of Michigan Cancer Foundation-7(MCF-7)cells.The 50%growth inhibitory concentration for MCF-7 cells at 24 h was 148.9 mmol/L.The proportions of MCF-7 cells arrested in the G0/G1 phase were 36.94+6.78%,45.06士5.60%,and 59.46+5.60%in the control group,75,and 150 mmol/L groups,respectively.Furthermore,we revealed that NGR1 treatment attenuates BC progression by targeted downregulating CCND2 and YBX3 genes.Additionally,YBX3 activates phosphatidylinositol 3-phosphate kinase(PI3K)/protein kinase B(Akt)signaling pathway by activating kirsten rat sarcoma viral oncogene,which is an activator of the PI3K/Akt signaling pathway.Conclusion:These results suggest that NGR1 can act as an efficacious drug candidate that targets the YBX3/PI3K/Akt axis in patients with BC.展开更多
目的研究miRNA-195在宫颈癌组织和细胞中的表达,分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法检测35例临床宫颈癌组织及其对应癌旁正常组织中miRNA-195,通过Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系;采用细胞...目的研究miRNA-195在宫颈癌组织和细胞中的表达,分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法检测35例临床宫颈癌组织及其对应癌旁正常组织中miRNA-195,通过Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系;采用细胞增殖实验和划痕迁移实验验证miRNA-195对宫颈癌siHa,Hela细胞增殖、迁移的影响;通过microRNA数据库预测miRNA-195的靶基因,双荧光素酶基因实验验证靶向结合关系;qRT-PCR验证miRNA-195对靶基因的调控;分析宫颈癌中靶基因的表达作用及与miRNA-195的相关性,通过细胞回补实验验证miRNA-195是否通过靶向调控蛋白表达在宫颈癌中发挥功能。结果宫颈癌组织中miRNA-195相对表达低于癌旁正常组织(21.03±5.17 vs 40.67±7.92),差异有统计学意义(t=12.285,P<0.001),且具有低表达预后差的临床特征(Logrank P=0.032)。过表达miRNA-195抑制了宫颈癌细胞的增殖(t=6.725~21.433,均P<0.01)和迁移速率(t=12.443,16.749,均P<0.001)。CCND2和MYB是miRNA-195的靶基因,过表达miRNA-195显著抑制了CCND2和MYB mRNA的蛋白表达(P<0.01)。宫颈癌组织中CCND2较癌旁正常组织显著高表达(52.67±4.79 vs 39.86±6.39),差异有统计学意义(t=12.453,P<0.001);MYB较癌旁正常组织显著高表达(43.06±6.43 vs 22.07±6.85),差异有统计学意义(t=13.217,P<0.001);且分别与miRNA-195表达呈负相关(r=-0.726,-0.592,均P<0.05)。过表达CCND2和MYB显著促进了宫颈癌细胞的增殖和迁移速率,敲低CCND2和MYB表达则得到与之相反的结果(F=144.947,875.160,均P<0.001);在过表达miRNA-195细胞中分别回补过表达CCND2和MYB后细胞增殖、迁移速率基本回归到正常水平。结论miRNA-195可通过靶向调控CCND2和MYB表达抑制宫颈癌癌细胞的增殖和迁移,进而参与宫颈癌的发生发展。展开更多
MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We in...MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We investigated miR-16 expression and their potential roles in a rat model of hypertrophy induced by abdominal artery constriction (AAC).miR-16 expression was significantly decreased, and CCND1 and CCND2 protein were markedly increased without obvious change of its mRNA level after hypertrophy induction.CCND1 and CCND2 levels were increased without changing their transcript levels in neonatal rat ventricular cardiomyocytes(NRVC) induced by PE,and miR-16 was down-regulated in this process with significantly up-regulatedβ-MHC,ANF and MLC-2 expression.Conversely,introduction of functional miR-16,CCND1 siRNA or CCND2 siRNA into NRVCs could repress cardiomyocyte hypertrophy.These results implicate that miR-16 is involved in contributing to cardiac hypertrophy,one of the mechanisms may be resulted from post-transcriptional regulation of CCND1 and CCND2.展开更多
基金This work was supported by grants from the National Natural Science Foundation of China(No.81260576)the Major Program of Science and Technology Foundation of Guizhou Province(QKHJZ[2014]2003)+1 种基金the Major Science and Technology Projects of Guizhou Province(QKHJZ[2015]2002-2-5)the Science and Technolo-gy Foundation of Guizhou Province(QKHJY[2011]3007).
文摘Background:Breast cancer(BC)is a common malignancy with highly female incidence.So far the function of notoginsenoside R1(NGR1),the extract from Panax notoginseng,has not been clearly elucidated in BC.Methods:Optimal culture concentration and time of NGR1 were investigated by cell counting kit-8 assay.Cell proliferation ability was measured by colony formation assays.Transwell assay was used to detect the effect of NGR1 on cell migration and invasion.The apoptosis rate of cells between each group was measured by TUNEL assay.Results:NGR1 treatment has an inhibitory efect on proliferation,migration,invasion,and angiogenesis and a stimulating effect on cell cycle arrest and apoptosis of Michigan Cancer Foundation-7(MCF-7)cells.The 50%growth inhibitory concentration for MCF-7 cells at 24 h was 148.9 mmol/L.The proportions of MCF-7 cells arrested in the G0/G1 phase were 36.94+6.78%,45.06士5.60%,and 59.46+5.60%in the control group,75,and 150 mmol/L groups,respectively.Furthermore,we revealed that NGR1 treatment attenuates BC progression by targeted downregulating CCND2 and YBX3 genes.Additionally,YBX3 activates phosphatidylinositol 3-phosphate kinase(PI3K)/protein kinase B(Akt)signaling pathway by activating kirsten rat sarcoma viral oncogene,which is an activator of the PI3K/Akt signaling pathway.Conclusion:These results suggest that NGR1 can act as an efficacious drug candidate that targets the YBX3/PI3K/Akt axis in patients with BC.
文摘目的研究miRNA-195在宫颈癌组织和细胞中的表达,分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法检测35例临床宫颈癌组织及其对应癌旁正常组织中miRNA-195,通过Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系;采用细胞增殖实验和划痕迁移实验验证miRNA-195对宫颈癌siHa,Hela细胞增殖、迁移的影响;通过microRNA数据库预测miRNA-195的靶基因,双荧光素酶基因实验验证靶向结合关系;qRT-PCR验证miRNA-195对靶基因的调控;分析宫颈癌中靶基因的表达作用及与miRNA-195的相关性,通过细胞回补实验验证miRNA-195是否通过靶向调控蛋白表达在宫颈癌中发挥功能。结果宫颈癌组织中miRNA-195相对表达低于癌旁正常组织(21.03±5.17 vs 40.67±7.92),差异有统计学意义(t=12.285,P<0.001),且具有低表达预后差的临床特征(Logrank P=0.032)。过表达miRNA-195抑制了宫颈癌细胞的增殖(t=6.725~21.433,均P<0.01)和迁移速率(t=12.443,16.749,均P<0.001)。CCND2和MYB是miRNA-195的靶基因,过表达miRNA-195显著抑制了CCND2和MYB mRNA的蛋白表达(P<0.01)。宫颈癌组织中CCND2较癌旁正常组织显著高表达(52.67±4.79 vs 39.86±6.39),差异有统计学意义(t=12.453,P<0.001);MYB较癌旁正常组织显著高表达(43.06±6.43 vs 22.07±6.85),差异有统计学意义(t=13.217,P<0.001);且分别与miRNA-195表达呈负相关(r=-0.726,-0.592,均P<0.05)。过表达CCND2和MYB显著促进了宫颈癌细胞的增殖和迁移速率,敲低CCND2和MYB表达则得到与之相反的结果(F=144.947,875.160,均P<0.001);在过表达miRNA-195细胞中分别回补过表达CCND2和MYB后细胞增殖、迁移速率基本回归到正常水平。结论miRNA-195可通过靶向调控CCND2和MYB表达抑制宫颈癌癌细胞的增殖和迁移,进而参与宫颈癌的发生发展。
文摘MicroRNAs(miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression post-transcriptionally. Recent studies have demonstrated that miRNAs are involved in the pathogenesis of hypertrophy.We investigated miR-16 expression and their potential roles in a rat model of hypertrophy induced by abdominal artery constriction (AAC).miR-16 expression was significantly decreased, and CCND1 and CCND2 protein were markedly increased without obvious change of its mRNA level after hypertrophy induction.CCND1 and CCND2 levels were increased without changing their transcript levels in neonatal rat ventricular cardiomyocytes(NRVC) induced by PE,and miR-16 was down-regulated in this process with significantly up-regulatedβ-MHC,ANF and MLC-2 expression.Conversely,introduction of functional miR-16,CCND1 siRNA or CCND2 siRNA into NRVCs could repress cardiomyocyte hypertrophy.These results implicate that miR-16 is involved in contributing to cardiac hypertrophy,one of the mechanisms may be resulted from post-transcriptional regulation of CCND1 and CCND2.
