Objective:MicroRNAs(miRNAs)are small,non-coding RNAs that play a key role in the development of chemoresistance in various cancer types,including colorectal cancer(CRC).In this study,we aimed to study the underlying m...Objective:MicroRNAs(miRNAs)are small,non-coding RNAs that play a key role in the development of chemoresistance in various cancer types,including colorectal cancer(CRC).In this study,we aimed to study the underlying mechanisms of miRNA in chemotherapy-resistant CRC.Methods:LoVo CRC cell line was exposed to oxaliplatin at an increased dose,and cells were cultured in the presence of oxaliplatin to develop LoVo^(OXR) cells.Microarray and Quantitative Reverse Transcription Polymerase Chain Reaction(qRT-PCR),western blot,and transwell assay were used to evaluate the chemoresistance in LoVo^(OXR) CRC cells.Results:Microarray and qRT-PCR analysis showed an increased expression of miR-100-5p in LoVo^(OXR) cells.MTT assay and flow cytometry analysis revealed less apoptosis and higher cell viability in LoVo^(OXR) cells.mRNA prediction target gene analysis showed C-terminal domain small phosphatase-like(CTDSPL),a phosphatase-like tumor suppressor,as a key target of miR-100-5p.CTDSPL expression was low in LoVo^(OXR) cells compared to LoVoWT cells.miR-100-5p regulates G1/S and S-phase transitions and inhibits differentiation by targeting the CTDSPL/pRB/E2F1 signaling pathway,which involves the modulation of cell cycle effectors in LoVo^(OXR) cells.Further,we found that forkhead box P3(FOXP3),as the upstream target of miR-100-5p,is highly expressed in LoVo^(OXR) cells.Inhibiting miR-100-5p and FOXP3 down-regulates miR-100-5p expression,while increased CTDSPL expression contributed to reduced cell proliferation and promoted cell apoptosis in LoVo^(OXR) CRC cells.Conclusions:miR-100-5p plays an oncogenic role in inducing chemoresistance through modulation of the CTDSPL/retinoblastoma protein(pRB)/E2F transcription factor 1(E2F1)axis in CRC cells.展开更多
【背景】酪蛋白水解蛋白酶L(caseinolytic protease L,ClpL)是热休克蛋白100(heat shock protein 100,HSP100)家族成员,参与多种细胞内生物学过程包括应激耐受反应、长期存活、毒力和抗生素抗性等,在病原菌的致病过程中发挥重要作用。...【背景】酪蛋白水解蛋白酶L(caseinolytic protease L,ClpL)是热休克蛋白100(heat shock protein 100,HSP100)家族成员,参与多种细胞内生物学过程包括应激耐受反应、长期存活、毒力和抗生素抗性等,在病原菌的致病过程中发挥重要作用。【目的】阐明HSP100/ClpL对猪链球菌2型(Streptococcus suis type 2)强毒株的应激耐受和毒力的影响。【方法】利用温敏自杀质粒pSET4s构建了SsClpL基因缺失株ΔclpL;利用大肠杆菌-猪链球菌穿梭质粒pSET2构建了互补株CΔclpL;比较亲本株SS2-1、ΔclpL和CΔclpL在各种应激环境(高温、氧化性应激和酸应激)和血液中的生存能力;利用小鼠感染模型和体内竞争试验评价clpL缺失前后细菌的毒力变化。【结果】clpL缺失后,SS2在各种应激环境中的生存能力明显减弱;同时在猪全血中的存活能力也显著降低;对BALB/c小鼠的致病力有所降低,在体内的定殖能力明显降低。【结论】SsClpL与猪链球菌的应激耐受和致病性密切相关。展开更多
基金supported by the China Medical University Hospital,Department of Medical Research(DMR-110-209)the Ministry of Science and Technology,Taiwan(MOST:109-2320-B-303-001-MY2),CMU107-ASIA-08,CMU109-MF-76 and CMU105-ASIA-01Taichung Armed Forces General Hospital:TCAFGH_D_115034.
文摘Objective:MicroRNAs(miRNAs)are small,non-coding RNAs that play a key role in the development of chemoresistance in various cancer types,including colorectal cancer(CRC).In this study,we aimed to study the underlying mechanisms of miRNA in chemotherapy-resistant CRC.Methods:LoVo CRC cell line was exposed to oxaliplatin at an increased dose,and cells were cultured in the presence of oxaliplatin to develop LoVo^(OXR) cells.Microarray and Quantitative Reverse Transcription Polymerase Chain Reaction(qRT-PCR),western blot,and transwell assay were used to evaluate the chemoresistance in LoVo^(OXR) CRC cells.Results:Microarray and qRT-PCR analysis showed an increased expression of miR-100-5p in LoVo^(OXR) cells.MTT assay and flow cytometry analysis revealed less apoptosis and higher cell viability in LoVo^(OXR) cells.mRNA prediction target gene analysis showed C-terminal domain small phosphatase-like(CTDSPL),a phosphatase-like tumor suppressor,as a key target of miR-100-5p.CTDSPL expression was low in LoVo^(OXR) cells compared to LoVoWT cells.miR-100-5p regulates G1/S and S-phase transitions and inhibits differentiation by targeting the CTDSPL/pRB/E2F1 signaling pathway,which involves the modulation of cell cycle effectors in LoVo^(OXR) cells.Further,we found that forkhead box P3(FOXP3),as the upstream target of miR-100-5p,is highly expressed in LoVo^(OXR) cells.Inhibiting miR-100-5p and FOXP3 down-regulates miR-100-5p expression,while increased CTDSPL expression contributed to reduced cell proliferation and promoted cell apoptosis in LoVo^(OXR) CRC cells.Conclusions:miR-100-5p plays an oncogenic role in inducing chemoresistance through modulation of the CTDSPL/retinoblastoma protein(pRB)/E2F transcription factor 1(E2F1)axis in CRC cells.
文摘【背景】酪蛋白水解蛋白酶L(caseinolytic protease L,ClpL)是热休克蛋白100(heat shock protein 100,HSP100)家族成员,参与多种细胞内生物学过程包括应激耐受反应、长期存活、毒力和抗生素抗性等,在病原菌的致病过程中发挥重要作用。【目的】阐明HSP100/ClpL对猪链球菌2型(Streptococcus suis type 2)强毒株的应激耐受和毒力的影响。【方法】利用温敏自杀质粒pSET4s构建了SsClpL基因缺失株ΔclpL;利用大肠杆菌-猪链球菌穿梭质粒pSET2构建了互补株CΔclpL;比较亲本株SS2-1、ΔclpL和CΔclpL在各种应激环境(高温、氧化性应激和酸应激)和血液中的生存能力;利用小鼠感染模型和体内竞争试验评价clpL缺失前后细菌的毒力变化。【结果】clpL缺失后,SS2在各种应激环境中的生存能力明显减弱;同时在猪全血中的存活能力也显著降低;对BALB/c小鼠的致病力有所降低,在体内的定殖能力明显降低。【结论】SsClpL与猪链球菌的应激耐受和致病性密切相关。
