Long noncoding RNA(lncRNA)IDH1 antisense RNA 1(IDH1-AS1)is involved in the progression of multiple cancers,but its role in epithelial ovarian cancer(EOC)is unknown.Therefore,we investigated the expression levels of ID...Long noncoding RNA(lncRNA)IDH1 antisense RNA 1(IDH1-AS1)is involved in the progression of multiple cancers,but its role in epithelial ovarian cancer(EOC)is unknown.Therefore,we investigated the expression levels of IDH1-AS1 in EOC cells and normal ovarian epithelial cells by quantitative real-time PCR(qPCR).We first evaluated the effects of IDH1-AS1 on the proliferation,migration,and invasion of EOC cells through cell counting kit-8,colony formation,EdU,transwell,wound-healing,and xenograft assays.We then explored the downstream targets of IDH1-AS1 and verified the results by a dual-luciferase reporter,qPCR,rescue experiments,and Western blotting.We found that the expression levels of IDH1-AS1 were lower in EOC cells than in normal ovarian epithelial cells.High IDH1-AS1 expression of EOC patients from the Gene Expression Profiling Interactive Analysis database indicated a favorable prognosis,because IDH1-AS1 inhibited cell proliferation and xenograft tumor growth of EOC.IDH1-AS1 sponged miR-518c-5p whose overexpression promoted EOC cell proliferation.The miR-518c-5p mimic also reversed the proliferation-inhibiting effect induced by IDH1-AS1 overexpression.Furthermore,we found that RNA binding motif protein 47(RBM47)was the downstream target of miR-518c-5p,that upregulation of RBM47 inhibited EOC cell proliferation,and that RBM47 overexpressing plasmid counteracted the proliferation-promoting effect caused by the IDH1-AS1 knockdown.Taken together,IDH1-AS1 may suppress EOC cell proliferation and tumor growth via the miR-518c-5p/RBM47 axis.展开更多
Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted ...Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.展开更多
目的研究肝细胞肝癌(HCC)中长链非编码RNA(LncRNA)氧化应激反应丝氨酸丰富1反义RNA1(OSER1-AS1)靶向调控微小RNA(miR)-433-3p的作用及生物学意义。方法选择唐山市第九医院手术切除的HCC组织和癌旁组织,培养HCC细胞株SMMC-7721、HepG2、M...目的研究肝细胞肝癌(HCC)中长链非编码RNA(LncRNA)氧化应激反应丝氨酸丰富1反义RNA1(OSER1-AS1)靶向调控微小RNA(miR)-433-3p的作用及生物学意义。方法选择唐山市第九医院手术切除的HCC组织和癌旁组织,培养HCC细胞株SMMC-7721、HepG2、MHCC97H及正常肝细胞株HL-7702。检测LncRNA OSER1-AS1、miR-433-3p的表达水平,比较HCC组织与癌旁组织、HCC细胞株与正常肝细胞株中LncRNA OSER1-AS1、miR-433-3p表达水平的差异。对转染MHCC97H细胞进行分组,转染阴性对照(NC)-siRNA为si-NC组,转染LncRNA OSER1-AS1-siRNA为si-OSER1-AS1组,转染LncRNA OSER1-AS1-siRNA及NC miR为si-OSER1-AS1+miR-NC组,转染LncRNA OSER1-AS1-siRNA及miR-433-3p抑制物为si-OSER1-AS1+miR-433-3p抑制物组。检测细胞增殖活力、凋亡率、增殖细胞核抗原(PCNA)及裂解型caspase-3(cleaved caspase-3)的表达水平,采用双荧光素酶报告基因实验验证LncRNA OSER1-AS1靶向miR-433-3p。结果HCC组织中LncRNA OSER1-AS1的表达水平高于癌旁组织(1.77±0.34 vs1.00±0.21)、miR-433-3p的表达水平低于癌旁组织(0.65±0.09 vs 1.00±0.28)(P<0.05)且LncRNA OSER1-AS1与miR-433-3p呈负相关(r=-0.351,P<0.05);HCC细胞株中LncRNA OSER1-AS1的表达水平高于HL-7702细胞、miR-433-3p的表达水平低于HL-7702细胞(P<0.05)且MHCC97H细胞中上述变化最显著。si-OSER1-AS1组MHCC97H细胞中LncRNA OSER1-AS1、PCNA的表达水平及增殖活力低于si-NC组(0.37±0.05 vs 1.00±0.11,0.33±0.05 vs 0.92±0.12,0.51±0.09 vs 1.03±0.12),miR-433-3p、cleaved caspase-3的表达水平及凋亡率高于si-NC组(1.88±0.25 vs 1.00±0.09,0.96±0.11 vs 0.44±0.06,9.39%±1.15%vs 3.82%±0.55%)(P<0.05);si-OSER1-AS1+miR-433-3p抑制物组MHCC97H细胞中cleaved caspase-3表达水平及凋亡率低于si-OSER1-AS1+miR-NC组(0.27±0.05 vs 0.91±0.10,6.04%±0.77%vs 11.32%±1.32%),PCNA的表达水平及增殖活力高于si-OSER1-AS1+miR-NC组(0.94±0.12 vs 0.48±0.06,0.95±0.11 vs 0.34±0.05)(P<0.05)。结论LncRNA OSER1-AS1靶向miR-433-3p调控HCC细胞的增殖和凋亡。展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.81572556 and 81402139).
文摘Long noncoding RNA(lncRNA)IDH1 antisense RNA 1(IDH1-AS1)is involved in the progression of multiple cancers,but its role in epithelial ovarian cancer(EOC)is unknown.Therefore,we investigated the expression levels of IDH1-AS1 in EOC cells and normal ovarian epithelial cells by quantitative real-time PCR(qPCR).We first evaluated the effects of IDH1-AS1 on the proliferation,migration,and invasion of EOC cells through cell counting kit-8,colony formation,EdU,transwell,wound-healing,and xenograft assays.We then explored the downstream targets of IDH1-AS1 and verified the results by a dual-luciferase reporter,qPCR,rescue experiments,and Western blotting.We found that the expression levels of IDH1-AS1 were lower in EOC cells than in normal ovarian epithelial cells.High IDH1-AS1 expression of EOC patients from the Gene Expression Profiling Interactive Analysis database indicated a favorable prognosis,because IDH1-AS1 inhibited cell proliferation and xenograft tumor growth of EOC.IDH1-AS1 sponged miR-518c-5p whose overexpression promoted EOC cell proliferation.The miR-518c-5p mimic also reversed the proliferation-inhibiting effect induced by IDH1-AS1 overexpression.Furthermore,we found that RNA binding motif protein 47(RBM47)was the downstream target of miR-518c-5p,that upregulation of RBM47 inhibited EOC cell proliferation,and that RBM47 overexpressing plasmid counteracted the proliferation-promoting effect caused by the IDH1-AS1 knockdown.Taken together,IDH1-AS1 may suppress EOC cell proliferation and tumor growth via the miR-518c-5p/RBM47 axis.
文摘Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.
文摘目的研究肝细胞肝癌(HCC)中长链非编码RNA(LncRNA)氧化应激反应丝氨酸丰富1反义RNA1(OSER1-AS1)靶向调控微小RNA(miR)-433-3p的作用及生物学意义。方法选择唐山市第九医院手术切除的HCC组织和癌旁组织,培养HCC细胞株SMMC-7721、HepG2、MHCC97H及正常肝细胞株HL-7702。检测LncRNA OSER1-AS1、miR-433-3p的表达水平,比较HCC组织与癌旁组织、HCC细胞株与正常肝细胞株中LncRNA OSER1-AS1、miR-433-3p表达水平的差异。对转染MHCC97H细胞进行分组,转染阴性对照(NC)-siRNA为si-NC组,转染LncRNA OSER1-AS1-siRNA为si-OSER1-AS1组,转染LncRNA OSER1-AS1-siRNA及NC miR为si-OSER1-AS1+miR-NC组,转染LncRNA OSER1-AS1-siRNA及miR-433-3p抑制物为si-OSER1-AS1+miR-433-3p抑制物组。检测细胞增殖活力、凋亡率、增殖细胞核抗原(PCNA)及裂解型caspase-3(cleaved caspase-3)的表达水平,采用双荧光素酶报告基因实验验证LncRNA OSER1-AS1靶向miR-433-3p。结果HCC组织中LncRNA OSER1-AS1的表达水平高于癌旁组织(1.77±0.34 vs1.00±0.21)、miR-433-3p的表达水平低于癌旁组织(0.65±0.09 vs 1.00±0.28)(P<0.05)且LncRNA OSER1-AS1与miR-433-3p呈负相关(r=-0.351,P<0.05);HCC细胞株中LncRNA OSER1-AS1的表达水平高于HL-7702细胞、miR-433-3p的表达水平低于HL-7702细胞(P<0.05)且MHCC97H细胞中上述变化最显著。si-OSER1-AS1组MHCC97H细胞中LncRNA OSER1-AS1、PCNA的表达水平及增殖活力低于si-NC组(0.37±0.05 vs 1.00±0.11,0.33±0.05 vs 0.92±0.12,0.51±0.09 vs 1.03±0.12),miR-433-3p、cleaved caspase-3的表达水平及凋亡率高于si-NC组(1.88±0.25 vs 1.00±0.09,0.96±0.11 vs 0.44±0.06,9.39%±1.15%vs 3.82%±0.55%)(P<0.05);si-OSER1-AS1+miR-433-3p抑制物组MHCC97H细胞中cleaved caspase-3表达水平及凋亡率低于si-OSER1-AS1+miR-NC组(0.27±0.05 vs 0.91±0.10,6.04%±0.77%vs 11.32%±1.32%),PCNA的表达水平及增殖活力高于si-OSER1-AS1+miR-NC组(0.94±0.12 vs 0.48±0.06,0.95±0.11 vs 0.34±0.05)(P<0.05)。结论LncRNA OSER1-AS1靶向miR-433-3p调控HCC细胞的增殖和凋亡。
