MTSS1基因在膀胱癌组织中的表达及对膀胱癌细胞增殖和凋亡的影响

目的研究MTSS1基因在膀胱癌中组织的表达及其对膀胱癌EJ138细胞增殖和凋亡的影响。方法荧光定量PCR检测41例膀胱癌及相应癌旁正常组织中MTSS1基因的表达。将MTSS1基因特异性的siRNA转染EJ138细胞。转染后,Western Blot检测转染后MTSS1蛋白的表达,MTT法测定细胞生长抑制率,流式细胞法检测细胞凋亡率。结果与癌旁正常组织相比,MTSS1 mRNA在膀胱癌组织中表达明显下调。RNA干扰能够显著抑制EJ138细胞中MTSS1蛋白的表达;MTSS1基因表达降低后,EJ138细胞的生长抑制率明显降低,细胞增殖活力明显增加,而且凋亡率明显降低。结论 MTSS1基因在膀胱癌低表达,促进膀胱癌细胞增殖并抑制细胞凋亡。     

miR-33a靶向MDM4对膀胱癌细胞增殖和凋亡的实验研究

目的 观察miR-33a对膀胱癌J82细胞增殖和凋亡的影响及可能机制.方法 采用Lipofectamine 3000将miR-33a mimics、miR-33a inhibitor、miR-33a negative转染至J82细胞,实时定量PCR方法验证转染效率,CCK-8和流式细胞术检测转染后J82细胞增殖能力和凋亡率变化,采用裸鼠成瘤和TUNEL方法检测miR-33a对膀胱癌J82细胞生长和细胞凋亡的影响.Western blot和荧光素酶实验检测miR-33a与膀胱癌J82细胞中调控鼠双微体4(MDM4)的关系.结果 实时定量PCR结果显示转染成功.miR-33a mimics能够抑制J82细胞增殖能力、促进凋亡,降低裸鼠成瘤体积与重量,升高凋亡指数(P＜0.01);miR-33ainibitor能够促进J82细胞增殖、抑制凋亡,增加裸鼠成瘤体积与重量,降低凋亡指数(P＜0.01).荧光素酶实验显示MDM4为miR-33a的下游靶基因,转染miR-33a抑制剂能够上调J82细胞MDM4的蛋白表达(P＜0.01).结论 miR-33a可直接靶向MDM4抑制J82细胞增殖、促进凋亡.     

大蒜素对人膀胱癌细胞BIU-87增殖和凋亡影响

目的:观察大蒜素对人膀胱癌细胞BIU-87增殖和凋亡的影响。方法:采用细胞计数法(Cell counting kit-8,CCK-8)检测不同浓度的大蒜素对BIU-87细胞生长的影响;流式细胞仪AnnexinV/PI双染法检测BIU-87细胞的凋亡率。结果:大蒜素作用于BIU-87细胞后,对其增值表现出了明显的抑制作用,且呈浓度依赖性;AnnexinV/PI双染法检测显示,不同浓度的大蒜素均可诱导BIU-87细胞凋亡,与阴性对照组比较,差异有统计学意义(P<0.05)。结论:大蒜素可抑制BIU-87细胞的增殖,诱导其凋亡。     

SEA诱导人膀胱癌BIU-87细胞凋亡机制的研究

目的：为了观察葡萄球菌肠毒素A(SEA)对人膀胱癌BIU-87细胞株诱导凋亡机制。方法：应用MTT比色法和流式细胞术检测SEA对BIU-87细胞增殖抑制率和对细胞周期的影响。结果：SEA对BIU-87细胞株有明显的抑制作用，且呈剂量依赖性。流式细胞术分析，SEA能阻止G1期细胞向S期的进程，G1期细胞堆积。结论：SEA能诱导BIU-87细胞凋亡，具有细胞毒性效应，可用于膀胱内灌注，从而抑制膀胱癌的浸润转移。     

miR-129-5p通过靶向Wnt5a抑制膀胱癌细胞的增殖和促进细胞凋亡

目的 检测miR-129-5p在膀胱癌组织和细胞中的表达及其对膀胱癌细胞的作用.方法 RT-qPCR法检测miR-129-5p在膀胱癌、癌旁组织以及膀胱癌细胞株中的表达.细胞中过表达miR-129-5p后,CCK-8实验检测细胞增殖能力变化;流式细胞术检测细胞凋亡情况;生物信息学网站预测miR-129-5p的靶点,并用...     

黄连素抑制人卵巢癌SKOV3细胞增殖并诱导凋亡

黄连素(berberine)对多种肿瘤有抑制增殖和诱导凋亡的作用[1-2],但在卵巢癌中的作用尚未见报道.本研究以人卵巢癌SKOV3细胞为研究对象,观察黄连素对SKOV3细胞增殖抑制作用及凋亡诱导效应.     

三氧化二砷抑制大鼠血管平滑肌细胞增殖并促进细胞凋亡

目的研究三氧化二砷(As2O3)对大鼠血管平滑肌细胞(VSMCs)的抑制增殖、促进凋亡及诱导细胞周期停滞的作用。方法经组织块贴壁法原代培养的大鼠VSMCs,应用MTT法、台盼蓝拒染法3、H-胸腺嘧啶胞苷掺入法测定VSMCs的细胞活力、生长及增殖;应用流式细胞仪、DNA梯带法分析细胞周期分布及凋亡;应用Western blot法检测凋亡相关基因产物P53及细胞周期蛋白激酶抑制蛋白P21waf1/cip1。结果1~16μmol/L As2O3对VSMCs细胞活力无明显影响,但可显著抑制细胞生长及DNA合成(P<0.05),并呈时间和剂量依赖性。8μmol/L As2O3可使细胞周期S期减少(P<0.05)、G0G1期增加(P<0.05),并出现sub-G1期凋亡峰。16μmol/L As2O3对VSMCs作用不同时间后出现典型的凋亡梯带样DNA片段。8μmol/L As2O3可显著上调VSMCs的P53及P21waf1/cip1蛋白产物(P<0.05),且呈时间依赖性。结论As2O3对VSMCs具有明确的抗增殖、促凋亡、阻滞细胞周期进程的作用,并且与P53、P21waf1/cip1表达上调密切相关。     

小檗碱增强阿霉素诱导的膀胱癌T24细胞凋亡

 目的:研究小檗碱对阿霉素诱导的膀胱癌T24细胞凋亡的影响及机制。方法:将膀胱癌T24细胞分为对照组、阿霉素组、阿霉素+小檗碱组和小檗碱组。采用CCK-8试剂盒测定膀胱癌T24细胞的增殖抑制率。采用Hoechst 33258染色剂检测细胞凋亡,同时测定caspase-3和caspase-9活性以及Bcl-2、Bax蛋白的表达。结果:小檗碱呈剂量和时间依赖性地促进阿霉素诱导的T24细胞凋亡。与阿霉素组比较,小檗碱+阿霉素组caspase-3、caspase-9活性和Bax蛋白的表达水平明显增加,而Bcl-2蛋白表达水平降低。结论:小檗碱可进一步增强阿霉素对T24细胞增殖的抑制作用,其机制与小檗碱增强阿霉素诱导的T24细胞凋亡有关。     

凋亡抑制蛋白在细胞凋亡中的调节作用

凋亡抑制蛋白(IAP)家族是至今发现的惟一一种内源性Caspase蛋白酶抑制剂,在细胞凋亡中发挥重要的调节作用。本文结合近年来的研究进展,综述了Caspase在凋亡中的作用、IAP家族蛋白分子的结构特征、抗凋亡作用机制以及IAPs抗凋亡作用的负调控。进一步认识细胞凋亡中复杂的调控机制,为寻找与凋亡紊乱相关的疾病治疗提供了重要靶向。     

三氧化二砷抑制兔血管平滑肌细胞增殖及诱导凋亡的相关机制

目的 研究三氧化二砷（As2O3）对免血管平滑肌细胞（VSMC）的抑制增殖、促进凋亡及诱导细胞周期停滞的作用.方法 经组织块贴壁法原代培养的兔VSMC,应用3H-胸腺嘧啶胞苷渗入法、透射电镜、TUNEL法、流式细胞术和基因芯片测定As2O3对VSMC的细胞活力、生长、增殖及凋亡.结果 随着三氧化二砷浓度的增加显著抑制了细胞生长,且呈剂量依赖性,透射电镜下可见凋亡小体,TUNEL检测出现了凋亡细胞,流式细胞术出现了典型的凋亡峰.基因芯片扫描杂交结果数据显示其中26条基因表达有差异.结论 As2O3对VSMC具有抑制增殖、促凋亡、阻滞细胞周期进程的作用.     

miR-503 inhibits cell proliferation and induces apoptosis in colorectal cancer cells by targeting E2F3

Objective: Colorectal cancer (CRC) is one of the major healthcare problems worldwide. A lot of miRNAs are aberrantly expressed in CRC and involved in its development and progression. The purpose of this study was to investigate the expression and function of miR-503 in CRC. Methods: miR-503 expression was detected in CRC tissues and cell lines by Quantitative real-time PCR. Cell proliferation was assessed by MTT assay. Cell apoptosis and cell cycle distribution were measured by flow cytometry. Moreover, luciferase reporter assay and western blot were performed to determine the potential target of miR-503 in CRC cells. Results: miR-503 was significantly decreased in CRC tissues and cell lines in comparison with controls. Overexpression of miR-503 in CRC cells remarkably inhibited cell proliferation and induced apoptosis. Furthermore, E2F3 was identified as a direct target of miR-503 in CRC cells and down-regulation of E2F3 had a similar effect as miR-503 overexpression on CRC cells. In addition, the expression of E2F3 was negatively correlated with miR-503 level in CRC tissues. Conclusions: miR-503 inhibits cell proliferation and induces apoptosis by directly targeting E2F3 in CRC cells, indicating its potential application in CRC diagnosis and therapy.     

PRAME induces apoptosis and inhibits proliferation of leukemic cells in vitro and in vivo

PRAME is a germinal tissue-specific gene that is expressed at high levels in haematological malignancies, but the physiological functions of PRAME in leukemia cells are unknown. It has reported that PRAME was found to be predominantly expressed in acute leukemias and high PRAME expression is correlated with a favorable prognosis in childhood acute leukemias, which suggested that PRAME could be involved in the regulation of cell death or apoptosis. In the present study, we tested a hypothesis that the PRAME gene plays a role in the regulation of apoptosis and proliferation of leukemia cells. We observed that KG-1 cells transient overexpressing the PRAME gene (when transfected with pcDNA3.1-PRAME plasmid) significantly induces apoptosis and decreases proliferation in vitro, and repression of PRAME expression by a short interfering RNA exhibited a increased proliferation in K562 cells in vitro and increases tumorigenicity of K562 leukemic cells in nude mice. Our results suggest that the leukemias expressing high levels of PRAME has favorable prognosis. PRAME may be as an attractive target for potential immunotherapy for acute leukemic.     

Osthole inhibits proliferation of human breast cancer cells by inducing cell cycle arrest and apoptosis

Recent studies have revealed that osthole,an active constituent isolated from the fruit of Cnidium monnieri(L.) Cusson,a traditional Chinese medicine,possesses anticancer activity.However,its effect on breast cancer cells so far has not been elucidated clearly.In the present study,we evaluated the effects of osthole on the proliferation,cell cycle and apoptosis of human breast cancer cells MDA-MB 435.We demonstrated that osthole is effective in inhibiting the proliferation of MDA-MB 435 cells,The mitochondrion-mediated apoptotic pathway was involved in apoptosis induced by osthole,as indicated by activation of caspase-9 and caspase-3 followed by PARP degradation.The mechanism underlying its effect on the induction of G1 phase arrest was due to the up-regulation of p53 and p21 and down-regulation of Cdk2 and cyclin D1 expression.Were observed taken together,these findings suggest that the anticancer efficacy of osthole is mediated via induction of cell cycle arrest and apoptosis in human breast cancer cells and osthole may be a potential chemotherapeutic agent against human breast cancer.     

过表达Rab27B体外促进人胃癌细胞系MKN-45凋亡并抑制其增殖

目的探讨Rab27B在胃癌细胞系MKN-45增殖和凋亡中的作用。方法构建Lv-Rab27B-EGFP表达载体并转染细胞作为实验组,以Lv-EGFP空载体转染细胞作为对照组,正常MKN-45作为空白组,CCK8测定细胞的增殖,流式细胞术检测细胞的周期和凋亡,Western blot检测凋亡相关蛋白Bax、Bcl-2和caspase3的表达。结果与阴性对照组和空白组细胞比较,实验组细胞的增殖受到抑制(P0.05),细胞周期显示有丝分裂受阻于G1期,出现凋亡增多(P0.05)。与对照组比较,实验组细胞促凋亡蛋白Bax表达量增加(P0.05),抑凋亡蛋白Bcl-2表达量显著下降(P0.05),caspase3蛋白也明显升高(P0.05)。结论 Rab27B过表达可以调节细胞周期而抑制人胃癌细胞系MKN-45增殖,要通过上调Bax和caspase3蛋白,以及下调Bcl-2蛋白表达来促进细胞凋亡。     

替米沙坦抑制U937细胞增殖及诱导凋亡

目的:探讨替米沙坦对U937细胞株的生长抑制及凋亡诱导作用。方法:分别以不同浓度的替米沙坦处理人类急性髓系白血病细胞U937;以CCK-8法检测不同浓度替米沙坦对U937细胞的生长抑制作用;以集落形成实验观察不同浓度替米沙坦对U937细胞集落形成能力的影响;以Annexin V-PI双染法及Hoechst 33342染色法检测不同浓度替米沙坦作用前后U937细胞凋亡程度的变化;以流式细胞术检测U937细胞表面抗原CD11b的阳性表达率,瑞氏染色后倒置显微镜进行细胞形态学观察,了解U937细胞的分化情况;以Western blot法检测不同浓度替米沙坦作用U937细胞后凋亡相关蛋白表达量的改变。结果:CCK-8实验结果证实替米沙坦呈时间和剂量依赖性抑制U937细胞的生长;集落形成实验显示低剂量替米沙坦可以完全抑制U937细胞的集落形成能力;Annexin V-PI双染法及Hoechst 33342法结果证实替米沙坦可以诱导U937细胞凋亡;细胞表面抗原流式检测术及瑞氏染色结果证实替米沙坦可以促进部分U937细胞分化;Western blot实验结果证实替米沙坦作用于U937细胞72 h后,促凋亡相关蛋白cleaved PARP及cleaved caspase-3蛋白的水平明显增高。结论:替米沙坦可以抑制细胞增殖以及诱导U937细胞部分分化,并通过caspase依赖的凋亡途径触发U937细胞凋亡。     

敲减microRNA-205靶向PTEN抑制食管癌细胞TE1的增殖并促进细胞凋亡

目的:探讨miRNA-205对食管癌细胞株TE1增殖和凋亡能力的影响及其作用机制.方法:实时定量聚合酶链式反应(qRT-PCR)和Western印迹检测正常食管黏膜细胞Het-1A和不同食管癌细胞株(KYSE70,TE12,TE1)中miRNA-205的mRNA及蛋白表达水平.转染miRNA-205抑制剂下调TE1细胞中miRNA-205的表达,采用CCK-8法和流式细胞术检测细胞增殖能力、细胞周期和细胞凋亡情况;Western印迹检测细胞增殖和细胞凋亡相关CDK2,cyclin D1,P21,Bcl-2,cleavedcaspase-3,caspase-3,p-Rb,Rb,p-Akt和Akt的蛋白表达水平.双荧光素酶报告基因分析法预测及验证其可能的靶基因.结果:MiRNA-205在各型食管癌细胞中高表达.沉默miRNA-205后,TE1细胞的增殖能力降低并表现为细胞周期阻滞,而细胞凋亡率显著升高(p＜0.05).同时细胞中cyclin D1,CDK2,bcl-2,p-Rb及p-Akt的蛋白表达水平均显著降低,p21及cleaved-caspase-3的蛋白表达水平显著升高.双荧光素酶报告基因分析显示PTEN是miRNA-205的可能作用靶点,TE1中共转染miRNA-205抑制剂和PTEN siRNA可部分逆转miRNA-205介导细胞增殖抑制及凋亡诱导作用.结论:沉默miRNA-205可靶向PTEN抑制食管癌细胞株TE1的增殖,并促进其凋亡,提示miRNA-205可作为食管癌诊疗的一个潜在作用靶点.     

Knockdown of NPM1 by RNA interference inhibits cells proliferation and induces apoptosis in leukemic cell line

Nucleophosmin (NPM1) is an abundant and ubiquitously expressed phosphoprotein that is known to influence solid tumors progression. However, little is known about the role of NPM1 in leukemia. Here, we knocked down the NPM1 expression by RNA interference to investigate the role of NPM1 in leukemic cells proliferation and apoptosis. The interference vector pNPM1-shRNA was constructed and transfected into the human leukemic K562 cell line. The expression levels of NPM1 mRNA and protein were detected by quantitative real-time PCR and Western blot, respectively. Cells proliferation potential in vitro was assessed by methyl thiazolyl tetrazolium (MTT) and colony formation assays. Flow cytometry was used to detect the distribution of cell cycle. Cellular apoptosis was reflected by the relative activities of caspase-3 and caspase-8. The results showed that the expression levels of NPM1 mRNA and protein in K562 cells were significantly reduced after pNPM1-shRNA transfection. The cells growth was significantly inhibited in a time-dependent manner and the number of colonies was significantly reduced in the pNPM1-shRNA transfected cells. Meanwhile, the percentage of cells in G1 phase in the K562/pNPM1-shRNA cells was significantly increased. In addition, there were higher relative activities of caspase-3/8 in the pNPM1-shRNA transfected cells. These results indicate that down-regulation of NPM1 expression inhibits leukemic cells proliferation, blocks cell cycle progression and induces cellular apoptosis. It may implicate a potential target for leukemia gene therapy.     

Rab5 GTPase对膀胱癌细胞自噬及增殖的调控

目的探讨Rab5 GTPase对膀胱癌细胞自噬及增殖的影响。方法采用实时定量PCR检测60例膀胱癌组织以及相应正常膀胱组织中Rab5 mRNA的表达水平,并检测Rab5 mRNA在膀胱癌细胞系及正常膀胱细胞系中的表达情况。通过pcDNA3.1-Rab5转染膀胱癌细胞系BIU-87、RT4过表达Rab5;免疫荧光、Western blot检测自噬相关蛋白LC3Ⅱ/Ⅰ、LC3斑点数目变化情况以及自噬相关蛋白Vps34的表达变化;采用CCK-8检测膀胱癌细胞的增殖改变情况。结果Rab5 mRNA在膀胱癌组织中的表达高于正常膀胱组织,Rab5 mRNA在膀胱癌细胞中的表达高于正常膀胱细胞系,差异有统计学意义(P<0.05);免疫荧光显示过表达Rab5后LC3荧光斑点明显增多,且过表达Rab5促进膀胱癌细胞中Vps34的表达增加及LC3-Ⅰ向LC3-Ⅱ的转化;过表达Rab5能促进膀胱癌细胞增殖(P<0.05)。结论膀胱癌组织中Rab5 mRNA的表达高于正常膀胱组织,Rab5提高膀胱癌细胞自噬水平并促进膀胱癌细胞增殖,且与自噬相关蛋白Vps34相关。