目的下调胶质瘤细胞系中CWF19L1(CWF19 like cell cycle control factor 1)的表达,探究胶质瘤细胞的增殖能力、迁移能力及侵袭能力的变化,为胶质瘤治疗研发新的治疗靶点。方法生物信息学分析肿瘤与正常组织中CWF19L1表达差异,验证胶质...目的下调胶质瘤细胞系中CWF19L1(CWF19 like cell cycle control factor 1)的表达,探究胶质瘤细胞的增殖能力、迁移能力及侵袭能力的变化,为胶质瘤治疗研发新的治疗靶点。方法生物信息学分析肿瘤与正常组织中CWF19L1表达差异,验证胶质瘤细胞中CWF19L1蛋白质的基础表达量,构建敲低慢病毒转染胶质瘤细胞,下调胶质瘤细胞中CWF19L1蛋白质表达。使用该稳转细胞系进行细胞周期、细胞增殖、细胞迁移和Transwell实验,以验证CWF19L1敲低对胶质瘤细胞功能的影响。结果生物信息分析表明肿瘤细胞中CWF19L1基因表达上调,蛋白质印迹法(WB)证明在U251胶质瘤细胞系中CWF19L1蛋白质表达量最高。敲低U251胶质瘤细胞中CWF19L1表达后,细胞周期实验示处于S/G2期的细胞比例上调(42.22%比33.00%P<0.05),CCK-8实验结果表明细胞数量明显上调(P<0.001),划痕实验显示划痕宽度明显缩短(P<0.001),迁移和侵袭实验证明穿过小室的细胞数量明显增多(P<0.001)。结论CWF19L1能够通过抑制细胞周期G1/S期转化,对胶质瘤细胞的增殖、迁移与侵袭能力产生抑制。展开更多
Pim-3 is a member of the provirus integration site for Moloney murine leukemia virus(Pim)family proteins that exhibit serine/threonine kinase activity.Similar to the other Pim kinases(Pim-1 and Pim-2),Pim-3 is involve...Pim-3 is a member of the provirus integration site for Moloney murine leukemia virus(Pim)family proteins that exhibit serine/threonine kinase activity.Similar to the other Pim kinases(Pim-1 and Pim-2),Pim-3 is involved in many cellular processes,including cell proliferation,survival,and protein synthesis.Although Pim-3is expressed in normal vital organs,it is overexpressed particularly in tumor tissues of endoderm-derived organs,including the liver,pancreas,and colon.Silencing of Pim-3 expression can retard in vitro cell proliferation of hepatocellular,pancreatic,and colon carcinoma cell lines by promoting cell apoptosis.Pim-3 lacks the regulatory domains similarly as Pim-1 and Pim-2 lack,and therefore,Pim-3 can exhibit its kinase activity once it is expressed.Pim-3 expression is regulated at transcriptional and post-transcriptional levels by transcription factors(e.g.,Ets-1)and post-translational modifiers(e.g.,translationally-controlled tumor protein),respectively.Pim-3 could promote growth and angiogenesis of human pancreatic cancer cells in vivo in an orthotopic nude mouse model.Furthermore,a Pim-3 kinase inhibitor inhibited cell proliferation when human pancreatic cancer cells were injected into nude mice,without inducing any major adverse effects.Thus,Pim-3 kinase may serve as a novel molecular target for developing targeting drugs against pancreatic and other types of cancer.展开更多
文摘目的下调胶质瘤细胞系中CWF19L1(CWF19 like cell cycle control factor 1)的表达,探究胶质瘤细胞的增殖能力、迁移能力及侵袭能力的变化,为胶质瘤治疗研发新的治疗靶点。方法生物信息学分析肿瘤与正常组织中CWF19L1表达差异,验证胶质瘤细胞中CWF19L1蛋白质的基础表达量,构建敲低慢病毒转染胶质瘤细胞,下调胶质瘤细胞中CWF19L1蛋白质表达。使用该稳转细胞系进行细胞周期、细胞增殖、细胞迁移和Transwell实验,以验证CWF19L1敲低对胶质瘤细胞功能的影响。结果生物信息分析表明肿瘤细胞中CWF19L1基因表达上调,蛋白质印迹法(WB)证明在U251胶质瘤细胞系中CWF19L1蛋白质表达量最高。敲低U251胶质瘤细胞中CWF19L1表达后,细胞周期实验示处于S/G2期的细胞比例上调(42.22%比33.00%P<0.05),CCK-8实验结果表明细胞数量明显上调(P<0.001),划痕实验显示划痕宽度明显缩短(P<0.001),迁移和侵袭实验证明穿过小室的细胞数量明显增多(P<0.001)。结论CWF19L1能够通过抑制细胞周期G1/S期转化,对胶质瘤细胞的增殖、迁移与侵袭能力产生抑制。
基金Supported by the National Science Foundation of China(in part),No.30973476 and No.812727
文摘Pim-3 is a member of the provirus integration site for Moloney murine leukemia virus(Pim)family proteins that exhibit serine/threonine kinase activity.Similar to the other Pim kinases(Pim-1 and Pim-2),Pim-3 is involved in many cellular processes,including cell proliferation,survival,and protein synthesis.Although Pim-3is expressed in normal vital organs,it is overexpressed particularly in tumor tissues of endoderm-derived organs,including the liver,pancreas,and colon.Silencing of Pim-3 expression can retard in vitro cell proliferation of hepatocellular,pancreatic,and colon carcinoma cell lines by promoting cell apoptosis.Pim-3 lacks the regulatory domains similarly as Pim-1 and Pim-2 lack,and therefore,Pim-3 can exhibit its kinase activity once it is expressed.Pim-3 expression is regulated at transcriptional and post-transcriptional levels by transcription factors(e.g.,Ets-1)and post-translational modifiers(e.g.,translationally-controlled tumor protein),respectively.Pim-3 could promote growth and angiogenesis of human pancreatic cancer cells in vivo in an orthotopic nude mouse model.Furthermore,a Pim-3 kinase inhibitor inhibited cell proliferation when human pancreatic cancer cells were injected into nude mice,without inducing any major adverse effects.Thus,Pim-3 kinase may serve as a novel molecular target for developing targeting drugs against pancreatic and other types of cancer.
