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
In this review, single-chain fragment variable construction using phage-display technology as a promising anticancer immunotherapy technology is described. Cloning and the specific bio-panning selection with phage dis...In this review, single-chain fragment variable construction using phage-display technology as a promising anticancer immunotherapy technology is described. Cloning and the specific bio-panning selection with phage display technology, as well as the use of the epidermal growth factor receptor (EGFR) at the surface of MCF-7 cells as the antigen for the straightforward specific selection of single chain Fvs, are discussed. Moreover, phage display technologies and their application are important for vaccine production and immunotherapy against viruses and cancers. Furthermore, expression of the gene will cause the production and expression of the protein in prokaryotic and eukaryotic cells, which can be used to detect anti-cancer single chain fragment variables (scFvs). Finally, homology modelling is described to show the three-dimensional scFv structure that verifies the Complementary-Determining-Regions (CDRs) on the surface of the model.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.
文摘目的探究血清嘌呤能离子通道型受体7(purinergic ligand-gated ion channel 7 receptor,P2X7R)、结缔组织生长因子(connective tissue growth factor,CTGF)表达与精神分裂症患者认知功能、临床症状的相关性。方法选取2021年1月~2023年1月在武汉市武东医院精神重症一科诊治的160例精神分裂症患者作为观察组,及同期160例健康体检志愿者作为对照组进行研究。根据阳性和阴性症状量表(positive and negative syndrome scale,PANSS)对患者精神临床症状(阳性和阴性症状、一般病理症状、附加症状)进行评估,将患者分为高分组(PANSS总分≥70分,n=72)和低分组(PANSS总分<70分,n=88)。利用精神分裂症认知功能成套测验共识版(MATRICS consensus cognitive battery,MCCB)评估患者认知能力;采用酶联免疫吸附测定(ELISA)法检测血清P2X7R和CTGF水平;Spearman法分析精神分裂症患者血清P2X7R和CTGF水平与PANSS各项评分、MCCB各项评分的相关性。结果与对照组相比,观察组血清P2X7R(610.71±107.83ng/L vs 384.78±80.62ng/L),CTGF水平(1.85±0.36μg/L vs 1.40±0.21μg/L)升高,差异有统计学意义(t=21.226,13.658,P<0.05);观察组精神分裂症患者MCBB各项评分均低于对照组,差异有统计学意义(t=14.845~24.862,均P<0.05);高分组精神分裂症患者阳性症状评分(21.10±3.42分)、阴性症状评分(23.37±5.03分)、一般病理症状评分(39.48±8.11分)、附加症状评分(8.26±1.22分)和PANSS总分(92.21±12.50分)均高于低分组(13.65±3.04分,15.62±3.91分,30.14±6.15分,5.20±0.94分,64.61±5.30分),差异有统计学意义(t=14.576,10.964,8.280,17.915,18.764,均P<0.05);高分组精神分裂症患者血清P2X7R,CTGF水平高于低分组,差异有统计学意义(t=12.233,5.923,均P<0.05);精神分裂症患者血清P2X7R,CTGF水平与PANSS各项评分均呈正相关(r=0.464~0.580,均P<0.05),与MCCB各项评分均呈负相关(r=-0.603~-0.439,均P<0.05)。结论精神分裂症患者血清P2X7R和CTGF水平升高,与患者临床症状和认知功能密切相关。
文摘In this review, single-chain fragment variable construction using phage-display technology as a promising anticancer immunotherapy technology is described. Cloning and the specific bio-panning selection with phage display technology, as well as the use of the epidermal growth factor receptor (EGFR) at the surface of MCF-7 cells as the antigen for the straightforward specific selection of single chain Fvs, are discussed. Moreover, phage display technologies and their application are important for vaccine production and immunotherapy against viruses and cancers. Furthermore, expression of the gene will cause the production and expression of the protein in prokaryotic and eukaryotic cells, which can be used to detect anti-cancer single chain fragment variables (scFvs). Finally, homology modelling is described to show the three-dimensional scFv structure that verifies the Complementary-Determining-Regions (CDRs) on the surface of the model.
