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
目的探究粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)通过多个C2结构域和跨膜区蛋白1反义RNA1(MCTP1 antisense RNA 1,MCTP1-AS1)/SMAD家族成员7(SMAD family member 7,SMAD7)表观遗传调控途径抑制上皮间质化转变...目的探究粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)通过多个C2结构域和跨膜区蛋白1反义RNA1(MCTP1 antisense RNA 1,MCTP1-AS1)/SMAD家族成员7(SMAD family member 7,SMAD7)表观遗传调控途径抑制上皮间质化转变(epithelial-mesenchymal transition,EMT)促进子宫内膜修复,并探讨其可能的潜在作用机理。方法通过qRT-PCR检测人宫颈内膜细胞H8细胞中G-CSF、E-钙黏蛋白(E-cadherin)、紧密连接蛋白Z0-1、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、SMAD7以及MCTP1-AS1的表达水平。MCTP1-AS1与G-CSF的相互作用采用双荧光素酶报告基因实验验证。结果G-CSF过表达可能抑制N-cadherin和Vimentin的表达,促进E-cadherin和Z0-1的表达(P<0.05);而在沉默G-CSF后可能促进了N-cadherin和Vimentin的表达,抑制E-cadherin和Z0-1的表达(P<0.05)。G-CSF靶向MCTP1-AS1。过表达的G-CSF可以抑制MCTP1-AS1和SMAD7的表达;相反,当G-CSF被敲除时,MCTP1-AS1和SMAD7的表达水平明显上调。结论通过细胞实验证实了G-CSF可能通过MCTP1-AS1/SMAD7表观遗传调控途径抑制上皮间质化转变促进子宫内膜修复,同时表明G-CSF可能是用于治疗薄型子宫的一个新靶点。展开更多
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.
文摘目的探究粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)通过多个C2结构域和跨膜区蛋白1反义RNA1(MCTP1 antisense RNA 1,MCTP1-AS1)/SMAD家族成员7(SMAD family member 7,SMAD7)表观遗传调控途径抑制上皮间质化转变(epithelial-mesenchymal transition,EMT)促进子宫内膜修复,并探讨其可能的潜在作用机理。方法通过qRT-PCR检测人宫颈内膜细胞H8细胞中G-CSF、E-钙黏蛋白(E-cadherin)、紧密连接蛋白Z0-1、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、SMAD7以及MCTP1-AS1的表达水平。MCTP1-AS1与G-CSF的相互作用采用双荧光素酶报告基因实验验证。结果G-CSF过表达可能抑制N-cadherin和Vimentin的表达,促进E-cadherin和Z0-1的表达(P<0.05);而在沉默G-CSF后可能促进了N-cadherin和Vimentin的表达,抑制E-cadherin和Z0-1的表达(P<0.05)。G-CSF靶向MCTP1-AS1。过表达的G-CSF可以抑制MCTP1-AS1和SMAD7的表达;相反,当G-CSF被敲除时,MCTP1-AS1和SMAD7的表达水平明显上调。结论通过细胞实验证实了G-CSF可能通过MCTP1-AS1/SMAD7表观遗传调控途径抑制上皮间质化转变促进子宫内膜修复,同时表明G-CSF可能是用于治疗薄型子宫的一个新靶点。
