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Use of rnai silencing to target preconditioned glial cell line-derived neurotrophic factor in neuronal apoptosis

Neural Regeneration Research 2011年第7期6卷 510-516页

作者： Hongliang Guo Xinhua Li Jing Mang Ying Xing Jinting He Guihua Xu Shijun Yan LifengLiu Chunli Mei Zhongxin Xu Department of Neurology China-Japan Friendship Hospital Jilin University Changchun 130031 Jilin Province China Department of Neurology Central Hospital of Changchun Changchun 130021 Jilin Province China Second Hospital of Jilin University Changchun 130021 Jilin Province China Beihua University Jilin 132001 Jifin Province China

Several studies have suggested that exogenous glial cell line-derived neurotrophic factor may pro-tect neurons from cerebral ischemic injury. However, the mechanisms underlying the neuroprotec-tive effects of endogenous glial cell line-derived neurotrophic factor remain unclear. The present experiments sought to elucidate the influence of various conditioned media on neuronal apoptosis, using a normal culture medium for astrocytes, an astrocyte medium highly expressing glial cell line-derived neurotrophic factor, and an astrocyte medium in which glial cell line-derived neurotro-phic factor expression was silenced using rnai technology. The results confirmed that the use of rnai silencing to target pretreated glial cell line-derived neurotrophic factor expression promoted neuronal apoptosis. In addition, oxygen and glucose deprivation preconditioning was found to upregulate glial cell line-derived neurotrophic factor expression, and significantly reduce neuronal apoptosis.

关键词： glial cell line-derived neurotrophic factor astrocyte neuron short interfering rna apoptosis neural regeneration

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Gene silencing of HIF-2α disrupts glioblastoma stem cell phenotype

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Cancer Drug Resistance 2020年第2期3卷 199-208页

作者： Leora M.Nusblat Shaili Tanna Charles M.Roth Department of Biomedical Engineering RutgersThe State University of New JerseyPiscatawayNJ 08854USA Department of Chemical and Biochemical Engineering RutgersThe State University of New JerseyPiscatawayNJ 08854USA

malignancy and recurrence in glioblastoma *** regions within the tumor microenvironment help maintain and promote the proliferation of ***,we explored the effects of silencing hypoxia inducible factor-2α(HIF-2α)because of its specificity for CSCs within the hypoxic ***:Cancer stem cell neurospheres were formed by enriching from both the glioblastoma cell line U87 and from brain tumor stem cells isolated directly from human brain *** of human HIF-2αwas performed using both commercial and in-house transfection of a validated short interfering rna,with all results compared to an established non-silencing control short interfering *** of HIF-2αwas established by Western blotting,and phenotypic effects were assayed by cell migration assays,cell viability measurements,and immunofluorescence staining of differentiation ***:Transfection with either our previously reported pH-sensitive,cationic amphiphilic macromolecule-based delivery system or Lipofectamine was similarly effective in silencing HIF-2α.The chemotherapeutic resistance and neurosphere formation were reduced when HIF-2αwas *** capacities in the presence of macrophage conditioned media were ***-2αsilencing was complementary to temozolomide treatment in producing phenotypic rather than cytotoxic ***:HIF-2αsilencing under hypoxia inhibited CSC phenotypes while promoting differentiated cell phenotypes and is complementary to existing DNA alkylating treatments in inhibiting glioma CSC activity.

关键词： Glioblastoma multiforme cancer stem cell short interfering rna tumor hypoxia

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Growth factor receptors: promising drug targets in cancer

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Journal of Cancer Metastasis and Treatment 2015年第1期1卷 190-200页

作者： Snigdha Tiash Ezharul Hoque Chowdhury Jeffrey Cheah School of Medicine and Health Sciences Faculty of MedicineNursing and Health SciencesMonash University46150 Bandar SunwaySelangorMalaysia

Genetic,epigenetic and somatic changes deregulate the expression of growth factor receptors(GFRs),leading to cancer initiation and *** cell growth and survival are orchestrated by clonal expansion and evasion of apoptotic signals in cancer *** growth of cells is further supported by angiogenesis and metastasis to distant *** expression of GFRs also contributes to the development of ***,therapeutics to target GFRs is a potentially attractive molecular approach to treat cancer more *** this review,we have discussed the contribution of GFRs to cancer development and addressed molecular approaches undertaken to inhibit GFR-mediated pathways.A wide number of monoclonal antibodies(mAbs)and protein kinase inhibitors targeting these GFR-mediated functions are in clinical trials to treat human ***,most drugs that target GFRs lead to the development of drug resistance and generate adverse *** acid-based therapeutics,*** interfering rna(sirna)could be harnessed to selectively silence GFR genes in cancer *** polymer,liposome-based nanocarriers,and the most recently developed pH-sensitive inorganic carbonate apatite nanoparticles have been used in cell culture and preclinical trials for cytoplasmic delivery of the sirnas targeting different GFR ***-based therapeutics have been shown to have signifi cant potential to suppress GFR expression and functions and thus could be developed as molecular ***-targeting of tumors at different levels by combining various approaches along with chemotherapy would be a promising therapeutic approach to fight the *** nanocarriers capable of entrapping sirna,mAb,GFR inhibitors and classical drugs targeting GFR have potential therapeutic applications.

关键词： Carbonate apatite nanoparticles growth factor receptor monoclonal antibodies protein kinase inhibitor short interfering rna tyrosine kinase inhibitor

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组蛋白去乙酰化酶在子宫颈癌靶向治疗中的作用研究

组蛋白去乙酰化酶在子宫颈癌靶向治疗中的作用研究

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作者：吴群英延边大学

学位级别：硕士

目的:探讨组蛋白去乙酰化酶(Histone deacetylase,HDAC)基因及其蛋白表达检测在宫颈癌及其癌前病变中的临床病理学意义以及HDAC抑制剂曲古抑菌素A(Trichostatin A,TSA)在宫颈癌分子靶向治疗中的作用。材料与方法:8例正常宫颈鳞状上皮,5... 详细信息

目的:探讨组蛋白去乙酰化酶(Histone deacetylase,HDAC)基因及其蛋白表达检测在宫颈癌及其癌前病变中的临床病理学意义以及HDAC抑制剂曲古抑菌素A(Trichostatin A,TSA)在宫颈癌分子靶向治疗中的作用。材料与方法:8例正常宫颈鳞状上皮,52例宫颈上皮内瘤变(Cervicalintraepithelial neoplasia,CIN),141例宫颈鳞状上皮细胞癌(Squamous cellcarcinomas,SCC)和24例宫颈腺癌(Adenocarcinomas,AC)共225例宫颈病变的存档蜡块标本选自延边大学医院、延边妇幼保健院和韩国高丽大学医科大学医疗中心病理科。同时,选取HeLa和CaSki两种宫颈癌细胞系作为TSA体外治疗效应的XTT检测对象。应用免疫组织化学、免疫荧光染色和Western Blot等实验方法检测5种HDAC基因蛋白在正常宫颈上皮、CIN和宫颈癌中的表达变化,并应用小分子rna干扰、XTT和流式细胞仪检测方法测定HDACs抑制剂TSA对于宫颈癌细胞的杀伤作用和机制。结果:HDAC1、3和HDAC5蛋白主要表现为细胞核阳性,在正常宫颈上皮中的表达率分别为0(0/8)、12.5%(1/8)和25%(2/8),而且阳性细胞位于上皮的基底细胞层,但在CIN、宫颈鳞癌和腺癌中的表达率明显升高。HDAC1在CIN、宫颈鳞癌和腺癌中阳性率分别为82.7%(43/50)、83%(117/141)和91.7%(22/24);HDAC3在CIN、宫颈鳞癌和腺癌中的阳性率分别为58.7%(27/46)、62.4%(87/141)和70.8%(17/24);HDAC5在CIN、宫颈鳞癌和腺癌中的阳性率分别为66%(33/50)、63.1%(89/141)和75%(18/24)。HDAC6则表现为胞浆阳性,在正常宫颈上皮中的阳性表达率为0(0/8),而且弱阳性染色细胞分布在上皮的基底部细胞层;在CIN、SCC和腺癌中的表达率也明显升高,分别达到84.6%(44/52)、83.0%(117/141)和91.7%(22/24)。HDAC2蛋白在宫颈正常上皮组织中表达为阴性,而且在宫颈癌及其癌前病变中的表达也较弱,只有少数病例表现为HDAC2蛋白阳性。TSA对于HeLaCaSki宫颈癌细胞具有明显的杀伤作用,但正常的皮肤鳞状上皮细胞对TSA则不敏感。而且,TSA处理组的HeLa细胞凋亡和死亡率明显较未经处理的HeLa对照组高,Acetylated Tubulin(乙酰化微管蛋白)的表达水平也明显上调。结论:HDAC1、3、5和HDAC6蛋白检测对于宫颈癌的诊断具有重要的辅助意义,而且可以作为宫颈癌治疗的新靶点。同时,HDAC抑制剂TSA对于宫颈癌具有明显的杀伤作用,其机制主要是通过上调Acetylated tubulin和诱导凋亡而实现的。

关键词： Histone Deacetylase 宫颈癌 short interfering rna 免疫组织化学免疫荧光染色 Western Blot

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