miR-142-5p对人上皮性卵巢癌细胞增殖和凋亡的影响

目的:探讨miR-142-5p靶向PTEN-PI3K/Akt信号调控人上皮性卵巢癌SKOV3细胞增殖和凋亡的具体机制。方法:Real-time PCR检测miR-142-5p在人卵巢癌组织及各卵巢癌细胞系中的表达情况。利用MTT、细胞克隆形成实验观察miR-142-5p对SKOV3细胞增殖活力的影响;Caspase-3活力检测miR-142-5p对SKOV3细胞凋亡情况;信号通路抑制剂处理及Western blot检测miR-142-5p、PTEN与PI3K/Akt信号通路的关系及PI3K/Akt信号下游基因Akt、FOXO1、cyclin D1等的表达情况;Luciferase实验验证miR-142-5p和PTEN 3' UTR的结合。结果:人上皮性卵巢癌组织及其细胞系中miR-142-5p表达显著增加（P<0.05)。与对照组相比,SKOV3细胞转染miR-142-5p mimics后活细胞数量显著增加,增殖能力显著上升,细胞凋亡下降（P<0.05);与之相反的,转染miR-142-5p inhibitor后SKOV3细胞增殖能力下调,凋亡增加。信号通路抑制剂处理显示miR-142-5p过表达激活PI3K/Akt信号通路。Luciferase实验显示miR-142-5p与PTEN 3' UTR直接结合。与对照组相比,miR-142-5p mimics组PTEN表达显著降低,p-Akt蛋白表达则显著上调（P<0.05）,同时过表达PTEN后p-Akt表达则显著降低（P<0.05）。结论:miR-142-5p通过抑制PTEN基因表达活化PI3K/Akt信号通路,促进人上皮性卵巢癌组织及SKOV3细胞增殖存活,抑制细胞凋亡。     

MiR-374a通过PTEN/AKT信号通路调节乳腺癌细胞的增殖和凋亡

摘 要：［目的］ 探讨miR-374a在乳腺癌细胞增殖和凋亡中的作用，并进一步探讨其机制。［方法］ 采用实时荧光定量PCR（qRT-PCR）检测人乳腺癌细胞MCF-7及正常人乳腺细胞Hs 578Bst的miR-374a及PTEN mRNA转录水平，Western blot法测定MCF-7、Hs 578Bst组细胞及对照、 miR-374a抑制剂组细胞的PTEN、p-AKT蛋白表达变化，用CCK-8法及Tunel法分别观察对照组和miR-374a抑制剂组细胞的增殖和凋亡，并用Western blot法检测两组细胞的凋亡相关蛋白Bcl-2、Cleaved caspase-3、Cleaved caspase-9表达。［结果］ 与人正常乳腺Hs 578Bst细胞相比，乳腺癌MCF-7细胞的miR-374a表达水平显著性增高（P<0.001），PTEN mRNA及蛋白的表达水平显著性降低（P<0.001），而p-AKT蛋白表达水平明显升高（P<0.001）。miR-374a抑制剂可抑制乳腺癌细胞的增殖（P<0.001），并促进其凋亡的发生（P<0.001），降低抗凋亡蛋白Bcl-2表达水平（P<0.001）的同时活化凋亡相关蛋白Caspase-3和Caspase-9（P<0.001）。miR-374a 通过靶点PTEN影响AKT通路，miR-374a组细胞PTEN mRNA表达水平显著性降低（P<0.001），而miR-374a抑制剂处理后，PTEN蛋白水平显著性升高（P<0.001），p-AKT表达降低（P<0.001）。［结论］ miR-374a在乳腺癌细胞高表达，并通过调节PTEN/AKT信号通路抑制乳腺癌细胞的增殖，促进凋亡的发生。miR-374a/PTEN/AKT信号通路有望成为乳腺癌的新治疗靶点。     

miR-429通过下调PTEN并激活PI3K/AKT信号通路促进胰腺癌PANC-1细胞对卡培他滨耐药

目的:探讨miR-429 靶向PTEN并通过PI3K/AKT信号通路调控胰腺癌PANC-1 细胞对卡培他滨的耐药性及其作用机制。方法：建立胰腺癌卡培他滨耐药细胞株PANC-1/CAP后,采用qRT-PCR和Western blotting 实验检测miR-429 和PTEN在胰腺癌细胞中的表达情况,平板克隆形成实验、CCK-8 法和Annexin V-FITC/PI 双染流式细胞术检测敲降miR-429 对胰腺癌卡培他滨耐药细胞株PANC-1/CAP 细胞增殖、凋亡和卡培他滨耐药性的影响,双荧光素酶报告基因验证miR-429 与PTEN 的靶向关系,Western blotting 实验进一步检测miR-429 对PTEN-PI3K/AKT信号通路的调控作用。结果：miR-429 在胰腺癌PANC-1 细胞和PANC-1/CAP细胞中的表达水平高于人胰腺导管上皮细胞（HPDE6-C7）（P<0.05 或P<0.01）,敲降miR-429 可显著抑制PANC-1/CAP细胞增殖活力、促进细胞凋亡及下调细胞卡培他滨耐药性,且双荧光素酶报告基因证实miR-429 靶向作用PTEN并下调其表达水平（P<0.05 或P<0.01）;敲降miR-429 可通过靶向上调PTEN并阻断PI3K/AKT信号通路进而显著抑制PANC-1/CAP细胞增殖活力,从而下调PANC-1/CAP细胞对卡培他滨的耐药性（P<0.05 或P<0.01）。结论：miR-429/PTEN-PI3K、AKT信号通路与胰腺癌卡培他滨耐药性存在调控关系,且敲降miR-429 可逆转PANC-1/CAP对卡培他滨的耐药性。     

靶向PTEN的肿瘤相关微小RNA

微小RNA (miRNA)是通过基因转录后沉默来调控基因表达的内源性非编码小RNA.其中miR-21、miR-17-92、miR-214、miR-26a、miR-221、miR-222等可以通过与PTEN mRNA 3'-UTR互补结合来抑制PTEN磷酸水解酶表达,而PTEN在细胞的增殖、凋亡、转移与侵袭中发挥重要作用...     

miR-103 靶向PTEN并激活PI3K/AKT通路促进肺癌细胞A549 对达沙替尼耐药

[摘要] 目的：探讨miR-103 靶向PTEN并激活PI3K/AKT信号通路促进肺癌细胞对达沙替尼（dasatinib,DASA）耐药的机制。方法：收集2014 年4 月至2018 年1 月昆明医科大学第一附属医院胸外科收治的资料完整的肺癌DASA耐药组织和不耐药组织各35 例。采用qPCR实验检测miR-103 在肺癌DASA耐药组织和细胞中的表达水平,同时,采用CCK-8、Transwell 和Wb实验检测敲降miR-103 对A549/DASA细胞增殖、迁移和上皮间质转化（EMT）的影响,双荧光素酶报告基因验证miR-103 与PTEN的靶向关系。进一步采用CCK-8、Transwell 和Wb实验检测miR-103 通过PTEN-PI3K/AKT信号通路对A549/DASA细胞恶性生物学行为的影响。结果：miR-103 在肺癌DASA 耐药组织和A549/DASA 细胞中均高表达（均P<0.01）。敲降miR-103 可显著抑制A549/DASA细胞的增殖、迁移和EMT（P<0.05 或P<0.01）。此外,双荧光素酶报告基因证实miR-103 靶向作用PTEN并下调其表达水平（P<0.01）。进一步实验显示,过表达miR-103 通过靶向下调PTEN并激活PI3K/AKT信号通路进而显著促进A549/DASA细胞增殖、迁移和EMT（P<0.05 或P<0.01）,从而上调A549/DASA细胞对DASA的耐药性。结论：miR-103/PTEN/PI3K/AKT信号通路与肺癌DASA耐药性存在调控关系,敲降miR-103 可逆转A549/DASA对DASA耐药。     

MircoRNA-214对吉非替尼敏感及耐药细胞增殖和凋亡的影响

摘 要：［目的］ 研究mircoRNA-214（miR-214）对PC9肺腺癌及吉非替尼耐药细胞（PC9/GR）增殖和凋亡的影响。［方法］ 在PC9细胞中转染miR-214模拟物及PC9/GR中转染miR-214抑制剂,使用定量逆转录PCR（qRT-PCR）检测其表达。MTT检测细胞转染miR-214模拟物或其抑制剂后的存活及增殖。在PC9细胞中顺时转染miR-214模拟物以检测上调miR-214对PC9细胞耐药性的影响,在PC9/GR细胞中顺时转染miR-214抑制物以检测下调miR-214对PC9/GR细胞耐药性的影响,并用流式细胞仪检测细胞的凋亡。Western blotting检测PTEN在PC9和PC9/GR细胞中的表达。构建PTEN 3’-UTR荧光素酶报告质粒验证miR-214的靶基因;建立异种移植模型检测miR-214抑制物对肺癌移植瘤的影响。［结果］ PC9细胞中miR-214低表达,上调miR-214的表达后PC9细胞对吉非替尼的敏感性降低,并且抵抗吉非替尼诱导的凋亡;而在PC9/GR细胞中低表达miR-214后,下调miR-214增加PC9/GR细胞对吉非替尼的敏感性,并且可以增强吉非替尼诱导的凋亡作用。荧光素酶报告载体实验证实PTEN是miR-214在细胞内的靶基因。动物异种移植模型表明miR-214抑制物可以增强PC9/GR对吉非替尼的敏感性。［结论］ MiR-214可能通过靶基因PTEN调控吉非替尼的获得性耐药。     

shRNA靶向沉默ZEB2表达通过Wnt/β-catenin信号通路降低肺癌细胞增殖活性的实验研究

目的:研究shRNA靶向沉默E盒结合锌指蛋白2（zinc finger E-box binding homeobox,ZEB2）表达对肺癌细胞增殖活性的影响。方法:用shRNA-ZEB2慢病毒和shRNA阴性对照慢病毒感染肺癌细胞,Real time PCR和Western blot检测沉默效果。MTT检测细胞增殖活性,流式细胞术检测细胞凋亡情况,Western blot检测细胞中激活型Caspase-3（Cleaved Caspase-3）、β-连环蛋白（β-catenin）、C-myc蛋白水平。用Wnt/β-catenin信号通路抑制剂处理沉默ZEB2的肺癌细胞,MTT检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中激活型Caspase-3、β-catenin、C-myc蛋白水平。结果:shRNA-ZEB2慢病毒可以明显沉默肺癌细胞中ZEB2的表达和转录,shRNA阴性对照慢病毒对肺癌细胞中ZEB2的表达没有影响。沉默ZEB2后的肺癌细胞增殖活性降低,细胞凋亡率升高,细胞中激活型Caspase-3蛋白水平升高,β-catenin、C-myc蛋白水平降低。Wnt/β-catenin信号通路抑制剂下调沉默ZEB2的肺癌细胞株中Wnt/β-catenin信号通路的激活水平,同时可以降低细胞存活率,诱导细胞凋亡,促进细胞中激活型Caspase-3的表达。结论:shRNA靶向沉默ZEB2表达通过Wnt/β-catenin信号通路降低肺癌细胞增殖活性。     

过表达miR-141-3p通过靶向下调PTEN并激活PI3K/Akt信号通路促进卵巢癌A2780 细胞的恶性生物学行为

[摘要] 目的：探讨miR-141-3p 通过靶向PTEN并调控PI3K/Akt 通路对卵巢癌细胞增殖、侵袭和凋亡的影响。方法：收集2014 年4 月至2017 年10 月河南省人民医院妇产科收治的资料完整的28 例卵巢癌患者肿瘤组织和相应的癌旁组织,采用qPCR检测卵巢癌组织和细胞系中miR-141-3p 的表达水平,双荧光素酶报告基因实验验证miR-141-3p 和PTEN的靶向关系;过表达或敲降miR-141 及PTEN基因后,采用CCK-8、Transwell 和Annexin V-FITC/PI 双染流式术检测卵巢癌A2780 细胞增殖、侵袭和凋亡水平,WB实验进一步检测miR-141-3p 对PTEN-PI3K/Akt 信号通路的调控作用。结果：miR-141-3p 在卵巢癌组织和细胞系中高表达（P<0.05 或P<0.01）。双荧光素酶报告基因证实miR-141-3p 靶向作用于PTEN并下调其表达水平（P<0.01）。与对照组相比,敲降miR-141-3p 后A2780 细胞的增殖受到显著抑制（48 h 时,0.36±0.04 vs 0.82±0.06,P<0.05）、侵袭能力明显降低[穿膜细胞数（45.14±7.88）vs（215.32±16.04）个,P<0.01]、细胞凋亡率显著升高[ （9.29±0.65）% vs（1.85±0.26）%,P<0.01]。过表达PTEN显著抑制了A2780 细胞中p-Akt 的表达（均P<0.01）、抑制细胞增殖和侵袭能力（均P<0.01）而明显促进细胞凋亡（均P<0.01）,在过表达PTEN的同时过表达miR-141-3p 或添加IGF-1 后可逆转上述的变化。结论：miR-141-3p 能够促进A2780 细胞增殖、侵袭和诱导凋亡,其机制可能与靶向调控PTEN并激活PI3K/Akt通路有关。     

沉默MCM7基因介导AKT信号通路参与黑色素瘤细胞增殖及凋亡的实验

目的 探讨MCM7基因沉默介导AKT信号通路参与人皮肤黑色素瘤细胞的增殖及凋亡。方法 构建MCM7基因和沉默MCM7基因表达的慢病毒RNA（LV-shRNA-MCM7）的表达载体;将A375细胞分为Control组、Empty vector转染组（空载质粒转染组）、siRNA（LV-shRNA-MCM7）转染组、siRNA NC（LV-shRNA-MCM7 negative control）转染组;MTT法检测每组细胞增殖;流式细胞术检测细胞周期、细胞凋亡情况;划痕实验法检测细胞迁移能力;qRT-PCR法测定细胞中MCM7基因、AKT信号通路相关基因、Cyclin D1及凋亡相关基因Bcl-2、Bax、caspase-3的相对表达量;Western blot法测定蛋白相对表达量。结果 沉默MCM7后,黑色素瘤细胞中的MCM7基因、CyclinD1、Bcl-2、AKT信号通路相关基因AKT3的mRNA和蛋白相对表达量显著下调（P<0.05）;凋亡相关基因Bax、caspase-3的mRNA和蛋白表达量显著上调（P<0.05）;siRNA转染组凋亡率显著上升,G₀/G₁期细胞数目明显增多,S期细胞数目明显减少;细胞增殖、迁移能力显著下降（P<0.05）。结论 沉默MCM7基因抑制AKT信号通路的激活,从而抑制A375黑色素瘤细胞增殖、迁移,促进A375黑色素瘤细胞凋亡。     

微小RNA-92a靶向PTEN／Akt信号通路对鼻咽癌细胞增殖与凋亡的影响

目的 探讨微小RNA-92a（miR-92a）通过靶向PTEN／Akt信号通路对鼻咽癌HONE1细胞增殖与凋亡的调控作用。方法 采用Lipofectamine脂质体法向HONE1细胞转染miR-92a抑制剂（miR-92a组）和阴性对照（NC组）,设不行转染的HONE1细胞为对照组。转染成功后用实时荧光定量PCR（QPCR）检测miR-92a在不同细胞组中的表达情况;MTT法检测各组细胞的增殖率;流式细胞技术检测各组细胞的凋亡率;Western blotting实验检测PTEN／Akt信号通路中PTEN和p-Akt蛋白的表达量。结果 转染48 h miR-92a组与对照组和NC组相比,miR-92a的表达量明显降低（P＜0.05）,而对照组和NC组的差异无统计学意义（P＞0.05）。miR-92a组转染24、48、72、96 h的细胞增殖率分别为（84.51±2.74）％、（77.21±3.55）％、（62.07±3.57）％和（49.25±4.15）％,其中72、96 h的细胞增殖率低于对照组和NC组（P＜0.05）;miR-92a组转染48 h的细胞凋亡率为（46.12±1.79）％,高于对照组的（6.99±0.72）％和NC组的（8.42±0.81）％,差异有统计学意义（P＜0.05）;miR-92a组转染48 h PTEN和p-Akt蛋白的表达水平分别为0.61±0.12和0.37±0.09,对照组分别为0.41±0.11和0.73±0.14,NC组分别为0.39±0.08和0. 68±0.07,与对照组和NC相比,miR-92a组细胞的PTEN表达升高,p-Akt表达降低,差异有统计学意义（P＜0.05）。结论 通过抑制miR-92a的表达来抑制鼻咽癌细胞增殖且促进其凋亡,可能是通过负调控PTEN／Akt信号通路来实现的。     

Downregulation of MicroRNA-147 Inhibits Cell Proliferation and Increases the Chemosensitivity of Gastric Cancer Cells to 5-Fluorouracil by Directly Targeting PTEN

Gastric cancer is the fourth most common malignancy and the third leading cause of cancer-related deaths worldwide. This study aimed to investigate the expression patterns, biological roles, and underlying mechanisms of microRNA-147 (miR-147) in gastric cancer. The present study demonstrated that miR-147 was significantly upregulated in gastric cancer tissues and cell lines. Downregulation of miR-147 decreased cell proliferation and enhanced the chemosensitivity of gastric cancer cells to 5-fluorouracil (5-FU) through the cell apoptosis pathway. In addition, phosphatase and tensin homolog (PTEN) was mechanically identified as the direct target of miR-147 in gastric cancer. PTEN knockdown reversed the effects of miR-147 downregulation on the proliferation, chemosensitivity, and 5-FU-induced apoptosis of gastric cancer cells. Moreover, miR-147 regulated the PI3K/AKT signaling pathway in gastric cancer by targeting PTEN. In conclusion, miR-147 suppressed the proliferation and enhanced the chemosensitivity of gastric cancer cells to 5-FU by promoting cell apoptosis through directly targeting PTEN and regulating the PI3K/AKT signaling pathway. This study provides important insight into the molecular mechanism that underlies the chemoresistance of gastric cancer cells. The results of this study could aid the development of a novel therapeutic strategy for gastric cancer.     

MicroRNA-106b promotes pituitary tumor cell proliferation and invasion through PI3K/AKT signaling pathway by targeting PTEN

The purpose of this study was to investigate the expression of microRNA-106b (miR-106b) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in pituitary tumor and to confirm whether miR-106b promotes proliferation and invasion of pituitary tumor cells through the PI3K/AKT signaling pathway by targeted regulation of PTEN expression, and thereby to find new targets for the treatment of pituitary tumor. Fifty-five cases of pituitary tumor tissue samples were collected, including 29 cases of invasive pituitary tumor, non-invasive 26 cases, and 8 normal pituitaries. The expression level of miR-106b in pituitary tumor tissue was detected by quantitative real-time PCR, and the expression of PTEN protein was detected by immunohistochemistry. PTEN 3′-untranslated region (UTR) luciferase vector was constructed, and dual-luciferase reporter gene assay was employed to examine the effect of miR-106b on PTEN 3′-UTR luciferase activity. AtT-20 cells were transfected with miR-106b mimics, miR-106b inhibitor, PTEN expression plasmid, and miR-106b mimics + PTEN expression plasmid respectively, and the changes in cellular proliferation and invasion were observed via MTT method and transwell assay respectively. PTEN messenger RNA (mRNA) expression was determined by quantitative real-time PCR, and western blotting was performed to detect the expression of PTEN, PI3K, AKT, and pAKT. miR-106b showed up-regulation in invasive pituitary tumor tissue: the expression level was significantly up-regulated compared with normal tissues and the non-invasive pituitary tumor tissue (P < 0.05). The positive rate of PTEN protein expression in invasive pituitary tumor tissues was significantly lower than in normal and non-invasive tissues (P < 0.01). Dual-luciferase reporter gene assay showed that miR-106b could bind to the 3ʹ-UTR of PTEN specifically and significantly inhibited the luciferase activity, cutting the 46 % (P < 0.01). Down-regulation of miR-106b or up-regulation of PTEN could suppress cell proliferation and invasion of AtT-20 cells, and PTEN expression plasmid could partially simulate the function of miR-106b. Expression of PTEN mRNA and protein decreased significantly in AtT-20 cells overexpressing miR-106b. The expression levels of PI3K and p-AKT were significantly inhibited by miR-106b inhibitor and increased by miR-106b mimics. The expression of miR-106b showed up-regulation in pituitary tumor tissues, while the protein expression of PTEN presented opposite results. The findings of this study further demonstrated that miR-106b as an oncogene regulated the pituitary tumor cell proliferation and invasion in vitro by directly targeting PTEN through the PI3K/AKT signaling pathway. Our study suggests that miR-106b and PTEN are likely to serve as potential diagnostic biomarkers or therapeutic targets for pituitary tumor treatment in the future.

     

miR-214 promotes tumorigenesis by targeting lactotransferrin in nasopharyngeal carcinoma

LTF (lactotransferrin, or lactoferrin) plays important role in innate immunity, and its anti-tumor function has also been reported in multiple cancers. We previously reported that LTF is significantly down-regulated in nasopharyngeal carcinoma (NPC) and acts as a tumor suppressor by suppressing AKT signaling. However, the exact mechanism of the down-regulation of LTF in NPC has not been revealed. In the current study, we screened and identified LTF is a bona fide target of miR-214 in NPC cells. miR-214 mimics significantly suppressed LTF mRNA and protein expression levels in NPC cells. miR-214 not only can promote NPC cell proliferation and invasion abilities in vitro, but also can accelerate tumor formation and lung metastasis in a mouse xenograft model. The pro-tumor function of miR-214 was depended on LTF suppression since LTF re-expression can reverse it. miR-214 can also activate AKT signaling by suppressing LTF expression. Furthermore, miR-214 expression level was up-regulated in NPC especially in metastasis-prone NPC tumor tissues compared with normal nasopharyngeal epithelial tissues, while the LTF expression level was negatively correlated with miR-214, suggesting that miR-214 targeting is partly responsible for LTF down-regulation in NPC specimens.     

MicroRNA-561 Affects Proliferation and Cell Cycle Transition Through PTEN/AKT Signaling Pathway by Targeting P-REX2a in NSCLC

MicroRNAs (miRNAs) play crucial roles in tumorigenesis and tumor progression. miR-561 has been reported to be downregulated in gastric cancer and affects cancer cell proliferation and metastasis. However, the role and underlying molecular mechanism of miR-561 in human non-small cell lung cancer (NSCLC) remain unknown and need to be further elucidated. In this study, we discovered that miR-561 expression was downregulated in human NSCLC tissues and cell lines. The overexpression of miR-561 inhibited NSCLC cell proliferation and cell cycle G1 /S transition and induced apoptosis. The inhibition of miR-561 facilitated cell proliferation and G1 /S transition and suppressed apoptosis. miR-561 expression was inversely correlated with P-REX2a expression in NSCLC tissues. P-REX2a was confirmed to be a direct target of miR-561 using a luciferase reporter assay. The overexpression of miR-561 decreased P-REX2a expression, and the suppression of miR- 561 increased P-REX2a expression. Particularly, P-REX2a silencing recapitulated the cellular and molecular effects observed upon miR-561 overexpression, and P-REX2a overexpression counteracted the effects of miR-561 overexpression on NSCLC cells. Moreover, both exogenous expression of miR-561 and silencing of P-REX2a resulted in suppression of the PTEN/AKT signaling pathway. Our study demonstrates that miR-561 inhibits NSCLC cell proliferation and G1 /S transition and induces apoptosis through suppression of the PTEN/ AKT signaling pathway by targeting P-REX2a. These findings indicate that miR-561 plays a significant role in NSCLC progression and serves as a potential therapeutic target for NSCLC.     

MicroRNA expression profiling in human ovarian cancer: miR-214 induces cell survival and cisplatin resistance by targeting PTEN

MicroRNAs (miRNA) represent a novel class of genes that function as negative regulators of gene expression. Recently, miRNAs have been implicated in several cancers. However, aberrant miRNA expression and its clinicopathologic significance in human ovarian cancer have not been well documented. Here, we show that several miRNAs are altered in human ovarian cancer, with the most significantly deregulated miRNAs being miR-214, miR-199a*, miR-200a, miR-100, miR-125b, and let-7 cluster. Further, we show the frequent deregulation of miR-214, miR-199a*, miR-200a, and miR-100 in ovarian cancers. Significantly, miR-214 induces cell survival and cisplatin resistance through targeting the 3'-untranslated region (UTR) of the PTEN, which leads to down-regulation of PTEN protein and activation of Akt pathway. Inhibition of Akt using Akt inhibitor, API-2/triciribine, or introduction of PTEN cDNA lacking 3'-UTR largely abrogates miR-214-induced cell survival. These findings indicate that deregulation of miRNAs is a recurrent event in human ovarian cancer and that miR-214 induces cell survival and cisplatin resistance primarily through targeting the PTEN/Akt pathway.     

食管鳞癌细胞外泌体中miR-130a对血管新生的作用及其机制

目的 探讨食管鳞癌细胞外泌体中的miR-130a对血管新生的作用及其机制.方法 提取食管上皮细胞HEEC、食管鳞癌细胞TE-13和Eca-109所分泌的外泌体及转染miR-130a mimic、miR-130a inhibitor及其阴性对照(NC)后的Eca-109细胞外泌体,并与人脐静脉血管内皮细胞HUVEC共孵育...     

Notch信号阻断剂抑制鼻咽癌细胞增殖的作用及机制

目的探讨Notch信号阻断剂抑制鼻咽癌细胞增殖的作用及其机制。方法应用Notch信号特异性抑制剂GSI抑制Notch信号的表达,MTT法检测癌细胞的生长增殖变化;流式细胞及Hoechst 33258染色检测癌细胞凋亡;应用Western blot检测Notch信号受抑制后AKT、MEK信号通路的变化。结果应用Notch抑制剂GSI作用后,癌细胞Notch1、2、4表达明显下降,而Notch3无明显变化。GSI作用后癌细胞增殖明显受到抑制,凋亡增加;该作用有浓度及时间双重依赖性（P<0.05）。Notch信号阻断后磷酸化AKT、GSK3β和ERK1/2显著下降,而总AKT、总GSK3β及总ERK1/2无明显变化。结论阻断鼻咽癌细胞Notch信号可以显著抑制鼻咽癌细胞增殖及诱导癌细胞凋亡。该作用可能与下调AKT及MEK信号通路有关。