As2O3诱导Raji细胞凋亡的基因分析

Objective The effect of arsenic trioxide on apoptosis gene expression of Raji cell was explored when Raji cells were incubated with 0.5μmol/L of arsenic trioxide for 6h。Methods Cell culture,extraction and isolation of mRNA,preparation of probes labeled with fluorescence,hybridization technique of DNA chip(each chip containing 200 apoptosis genes,Chinese Shanghai Biostar,In.)were used.Results Arsenic trioxide induced significant changes in 10%(20/200 genes)of the apoptosis genes:18 genes were downregulated,only two upregulated.In particular,inhibitors of apoptosis protein,such as X-linked inhibitor of apoptosis protein,were significantly downregulated.P53 and the other apoptosis genes were also downregulatec.Of the upregulated genes,high expression of heat-shock protein could promote apoptosis of Raji cells.Conclusion The inhibitors of apoptosis protein play an important role in the process of arsenic trioxide-induced apoptosis of Raji cells.     

三氧化二砷(As2O3)诱导人肺腺癌GLC-82细胞凋亡及分子机制的研究

目的:探讨As2O3对人肺腺癌GLC-82细胞系抑制生长、诱导凋亡的生物学效应及作用机制.方法:通过MTT还原法检测As2O3对该细胞系生长的影响;用光学显微镜、流式细胞仪、DNA凝胶电泳和细胞凋亡原位检测(TUNEL)研究As2O3诱导细胞凋亡的情况;用RT-PCR、Northern blot或Western blot分析As2O3对c-myc、p53、p16和bcl-X等基因表达的影响.结果:As2O3能抑制GLC-82细胞的生长.As2O3处理GLC-82细胞后,光学显微镜下可见到明显的凋亡细胞,细胞周期的G1期前有低于2倍体的凋亡峰,DNA凝胶电泳显示出典型的凋亡特征:DNA有规律断裂形成的梯状图谱,蛋白水平的检测表明As2O3可使细胞c-myc基因表达下降,p16和p53表达升高.结论:As2O3能显著抑制GLC-82细胞生长、诱导细胞凋亡,并主要通过调节c-myc,p16和p53等基因的表达来实现.     

As2O3对人胃癌细胞株SCG-7901裸鼠移植瘤的抗肿瘤机制研究

目的：探讨不同剂量的三氧化二砷（As2O3）对人胃癌细胞株SCG-7901裸鼠皮下移植瘤的抗肿瘤机制。方法：建立人胃癌裸鼠皮下移植瘤模型,随机分为实验组即As2O3低剂量组（1．0mg／kg）、As2O3中剂量组（2．5mg／kg）和As2O3高剂量组（5．0mg／kg）,空白对照生理盐水组及阳性对照5-Fu组（20mg／kg）,分别腹腔连续给药10天,停药后24小时处死裸鼠,称取瘤重,计算抑瘤率,流式细胞仪检测凋亡率及Caspase-3的活性变化。结果：低剂量As2O3组及5-FU组抑瘤率分别为60．6％及78．2％,与生理盐水组比较均有统计学意义（P〈0．05）,低剂量组与5-FU组抑瘤率差异无统计学意义（P〉0．05）;高、中、低剂量As2O3组流式细胞仪检测时均出现典型凋亡峰,凋亡率分别为18．32％、17．86％、21．89％,同时伴有Caspase-3活性的增加,其中低剂量As2O3活性增加最显著。结论：低剂量As2O3在体内可通过活化Caspase-3诱导细胞凋亡而发挥抗肿瘤作用。     

As2O3诱导胰腺癌细胞凋亡的相关研究

目的探讨三氧化二砷(arsenictrioxide,As2O3)对胰腺癌细胞的生长的影响及其作用机制。方法采用不同浓度的As2O3处理胰腺癌AsPC-1细胞不同时间(1d、2d、3d),在显微镜和电镜下观察细胞的形态,噻唑蓝(MTT)法和流式细胞分析法测定细胞生长抑制情况;不同浓度的As2O3处理细胞2h、5h、8h、12h、24h后,用DCFH-DA标记细胞后检测荧光强度反映细胞内过氧化氢(H2O2)水平。结果1μmol/LAs2O3处理AsPC-1细胞24h即呈现明显的生长抑制和凋亡特征。As2O3对细胞内H2O2水平影响的结果显示As2O3处理AsPC-1细胞2h后,细胞内H2O2的水平即明显升高(10μmol/L组达79%),到5h达到高峰(增高169%),12h后下降,24h水平与对照组接近,且H2O2水平的变化对As2O3有明显的剂量依赖性。结论As2O3能诱导AsPC-1细胞凋亡,其机制可能与细胞内的H2O2水平改变有关,并且H2O2积累是As2O3诱导细胞凋亡途径中的早期事件,H2O2可能在As2O3诱导肿瘤细胞凋亡途径中扮演类似第二信使的角色。     

三氧化二砷抗肝癌机制及其在肝癌治疗中的研究

三氧化二砷(ATO)参与肝癌细胞的多种生物学过程,包括肿瘤细胞凋亡诱导、增殖抑制、侵袭转移抑制和肿瘤干细胞抑制等。ATO在肝癌治疗中有多种方式,主要包括单药治疗、联合局部治疗、联合全身治疗等。进一步研究ATO抗癌机制及临床应用有望为肝癌治疗提供新的思路。     

As2O3联合Ad-pml体外抗肝癌细胞作用的实验研究

目的:探讨三氧化二砷(As2O3)联合腺病毒介导的早幼粒细胞性白血病基因(promyelocytic leukemia-adenovirus,Ad-pml)体外抗肝癌细胞作用。方法:同时设Ad-pml组、As2O3组、As2O3联合Ad-pml组以及正常细胞对照组,运用分子生物学和细胞生物学技术,观察联合应用As2O3和Ad-pml在体外治疗肝癌的效果,并进一步探讨其作用机制及特点。结果:Ad-pml与As2O3对肝癌细胞体外生长的影响:两因素析因设计分析结果表明,单用Ad-pml处理肝癌细胞,其抑制作用随Ad-pml MOI值的增加而增加(P<0.05),单独应用As2O3在浓度小于5μmol/L时,未发现有明显的肿瘤抑制作用(P>0.05);As2O3和Ad-pml联合应用具有协同抗肿瘤作用,二者之间存在着交互作用(P<0.05)。流式细胞仪检测细胞凋亡结果:用Ad-pml和As2O3共同作用肝癌细胞24小时后,早期细胞凋亡率明显增高。RT-PCR结果表明,联合治疗组p53表达最高,而Bcl-2表达最低,相反,肝癌细胞对照组p53表达最低,而Bcl-2表达最高。结论:As2O3和Ad-pml联合应用有协同抗肝癌细胞作用,其机制与增强肝癌细胞凋亡及控制凋亡相关基因的表达有关。     

As2O3通过下调Annexin Ⅱ抑制NB4细胞侵袭能力的研究

目的:旨在探讨三氧化二砷(arsenic trioxide ,As2O3)通过下调钙依赖性磷脂结合蛋白Annexin Ⅱ(AnxA2)抑制人急性早幼粒白血病(acute promyelocytic leukemia,APL)细胞NB4侵袭力的作用.方法:体外培养NB4细胞,FCM检测不同浓度As2O3对NB4细胞膜表面AnxA2蛋白表达的影响;Transwell法检测As2O3和AnxA2抗体对NB4细胞侵袭力的影响.结果:As2O3在0.31、0.63和 1.25 μg/mL质量浓度条件下均能诱导NB4细胞膜表面AnxA2蛋白水平表达下降(P<0.05),呈剂量依赖性;AnxA2抗体质量浓度为0.01、0.02和0.04 μg/mL以及AS2O3质量浓度为0.31、0.63和1.25 μg/mL条件下均能使NB4细胞的侵袭力下降(P<0.05),并呈剂量依赖关系. 结论:AnxA2抗体可抑制NB4细胞的侵袭力,As2O3可通过下调AnxA2蛋白在细胞膜表面的表达,从而抑制NB4细胞的侵袭能力.     

As2O3联合L-OHP对HepG2肝癌细胞株的体外抑制作用

目的探讨As2O3联合L-OHP对HepG2肝癌细胞株的体外抑制作用。方法采用MTT法（四唑盐比色法）动态观察As2O3联合L-OHP对HepG2肝癌细胞的生长抑制作用,并采用两药相互作用系数（CDI）来评价两药相互作用的性质;应用流式细胞术（FCM）检测凋亡率和细胞周期分布。结果 As2O3与L-OHP联合应用,对HepG2肝癌细胞株的生长抑制、诱导凋亡作用均较相应的单药增强。流式细胞直方图上可见明显的＂凋亡峰＂,且As2O3使肝癌细胞周期阻滞于G2/M期,L-OHP使细胞周期阻滞于S期和G2/M期,两药合用使细胞周期阻滞于S期和G2/M期。结论中低浓度As2O3与L-OHP联合作用具有显著的协同效益,其主要机制可能是As2O3联合L-OHP诱导肝癌细胞凋亡作用及增强细胞周期阻滞作用。     

三氧化二砷对人肝癌HepG2细胞内活性氧水平的影响

[目的]探讨三氧化二砷（As2O3）对人肝癌细胞生长抑制和诱导凋亡作用及其对细胞内活性氧（ROS）水平的影响。[方法]用MTT法、DNALadder检测及流式细胞术等观察As2O3对人肝癌细胞株HepG2的生长抑制和诱导凋亡作用及细胞内ROS的变化。[结果]As2O3能明显抑制HepG2细胞的增殖,并呈时间和剂量依赖性。DNALadder检测示,121μmol／L As2O3处理HepG2细胞48h开始出现DNALadder条带。流式细胞仪分析显示As2O3处理人肝癌细胞HepG2 12、24、36h后细胞内ROS水平较对照组明显升高（P〈0．01）,且呈时间依赖性。[结论]As20,可通过抑制人肝癌细胞HepG2增殖和提高细胞内ROS水平诱导细胞凋亡而发挥抗肿瘤作用。     

抑制端粒酶活性对As2O3诱导肝癌细胞凋亡的影响

目的： 〖HT5"SS〗观察端粒酶反义寡核苷酸、As2O3对肝癌细胞生长的影响,寻找低剂量、低毒、高效、安全的抗肝癌新疗法。〖HT5W〗方法：〖HT5"SS〗 自行设计合成靶向端粒酶模板区的20 nt硫代反义寡核苷酸,观察端粒酶反义寡核苷酸与As2O3对肝癌细胞端粒酶活性的影响及两者协同对肝癌细胞生长的影响,采用HE染色、流式细胞仪及DNA琼脂糖凝胶电泳观察端粒酶反义寡核苷酸与As2O3对肝癌细胞的凋亡诱导作用,以流式细胞术检测肝癌细胞的Fas、FasL、bcl2蛋白的表达。〖HT5W〗结果： 〖HT5"SS〗5 μmol/L的端粒酶反义寡核苷酸作用24h可显著抑制肝癌细胞端粒酶活性（P<0.01）;肝癌细胞端粒酶活性被抑制后对As2O3诱导凋亡的敏感性显著增加,表现为低浓度、短时间即可诱导肝癌细胞凋亡（P<0.01）,端粒酶反义寡核苷酸与As2O3对肝癌细胞的凋亡诱导作用是通过Fas、FasL途径实现的。〖HT5W〗结论：〖HT5"SS〗抑制肝癌细胞端粒酶活性可显著增强其对As2O3诱导凋亡的敏感性,减少砷剂用量,两者协同有潜在的临床应用前景。     

RNA干扰SMYD3基因表达对诱导肝癌细胞凋亡的影响

背景与目的:SMYD3(SETandMYNDdomain-containingprotein3)基因的表达蛋白是一种组蛋白甲基转移酶,参与肿瘤细胞增殖与凋亡的调控。本研究旨在探讨利用RNA干扰(RNAinterference,RNAi)抑制SMYD3基因表达对肝癌细胞增殖与凋亡的影响。方法:RT-PCR、免疫组织化学法分别检测SMYD3在肝癌细胞和肝癌组织中的表达。构建小发夹状RNA(smallhairpinRNA,shRNA)干扰质粒Pgenesil-1-s1、Pgenesil-1-s2及无干扰效应质粒Pgenesil-1-hk并转染入肝癌细胞HepG2,阻抑其表达SMYD3,以空质粒Pgenesil-1转染组为对照。Westernblot检测阻抑效应;MTT检测细胞增殖抑制率,流式细胞术及TUNEL检测细胞凋亡。结果:SMYD3在肝癌组织和多种肝癌细胞中表达明显增强。shRNA转染HepG2细胞后:SMYD3蛋白表达下调75%～85%;细胞增殖明显受抑制,抑制率高达60.95%～72.14%;流式细胞术结果显示Pgenesil-1-s1组HepG2细胞凋亡率(17.68±2.36)%、Pgenesil-1-s2组(19.07±1.78)%,均显著高于Pgenesil-1-hk组[(1.44±0.28)%]及Pgenesil-1组[(0.47±0.12)%](P<0.01);TUNEL检测的凋亡指数结果与流式细胞术检测结果类似。结论:SMYD3高表达于多种肝癌细胞及肝癌组织;RNAi能特异性下调SMYD3的表达,抑制肝癌细胞增殖并促进细胞凋亡,提示其可能为治疗肝癌提供新的途径。     

Significance of COX-2 expression in human renal cell carcinoma cell lines

Accumulating evidences indicate that cyclooxygenase (COX)-2 plays an important role in tumorigenesis in many human cancers. Yet the relationship between COX-2 and human renal cell carcinoma (RCC) remains unclear. The aim of our study was to evaluate COX-2 expression in human RCC cell lines and its role in tumorigenesis of human RCC. Among the human RCC cell lines (SMKT-R4, OS-RC-2, ACHN) and normal renal cell line RPTEC, COX-2 overexpression was found in OS-RC-2 cells both at mRNA and protein levels. COX-2 sense- and antisense-orientated vectors were constructed and transferred into RCC cells. Significant suppression of cellular proliferation was demonstrated in OS-RC-2 antisense transfectants, whereas promotion was found in SMKT-R4 sense transfectants by colony-forming assay despite the observation that COX-2 specific inhibitor NS398 exhibited similar IC50 among RCC cell lines by MTT assay. In comparison with parent cells and sense transfectants, significant suppression of COX-2 expression and PGE2 production and increase in butyrate-induced apoptosis were observed in OS-RC-2 antisense transfectants by Western blot, ELISA assay and FACS analysis, respectively. Furthermore, tumor growth and angiogenesis of OS-RC-2 antisense transfectants in nude mice was significantly suppressed and the survival time of these mice was significantly prolonged. Our study demonstrates that COX-2 is overexpressed in OS-RC-2 RCC cell line and plays an important role in tumorigenesis of the cells in vivo, which implies that COX-2 may be a therapeutic target for COX-2-expressing RCC, and that suppression of COX-2 expression by antisense-based strategy may have potential utility in treatment of COX-2-expressing RCC.     

Significance of COX-2 expression in human esophageal squamous cell carcinoma

Cyclooxygenase-2 (COX-2) is well established to play an important role in the tumorigenesis of a variety of human cancers; however, the function of COX-2 in the development of esophageal squamous cell carcinoma (ESCC) remains less clear. Here, we determined, first, the pattern of COX-2 expression in normal esophageal mucosa, dysplasia, carcinoma in situ (CIS) and invasive SCC. Immunohistochemical analysis showed that, while COX-2 was weakly expressed, if at all, in normal squamous epithelium, strong COX-2 expression was detected as early as the stage of dysplasia and frequently in 20 of 26 (77%) CIS and 86 of 111 (77%) invasive SCC. Upregulation of COX-2 in ESCC was found to be significantly associated with tumor progression (R = 0.493, P < 0.01). Further, treatment of human ESCC cell lines (KYSE450 and KYSE510) with NS-398, a COX-2 specific chemical inhibitor, suppressed the production of prostaglandin E2 (PGE2) and induced cell growth inhibition, cell cycle arrest at the G1-S checkpoint, and the expression of cyclin-dependent kinase inhibitors p21waf1/cip1 and p27kip1. Finally, knockdown expression of COX-2 in KYSE450 cells by a specific COX-2 siRNA dramatically inhibited PGE2 production, cell growth and, more importantly, colony formation and tumorigenesis in nude mice. Together, this study suggested that COX-2 may be involved in an early stage of squamous cell carcinogenesis of the esophagus and has a non-redundant role in the regulation of cellular proliferation and tumorigenesis of esophageal epithelial cells.     

番荔枝内酯Bullatacin对人肝癌细胞株HpG2凋亡影响的探讨

目的:研究番荔枝内酯 Bullatacin在诱导人肝癌细胞株HpG2凋亡中的作用.方法:MTT法测定Bullatacin对HpG2细胞增殖的影响;采用碘化丙锭染色(PI)及荧光标记的单克隆抗体(mAb)结合流式细胞仪(FCM)分析Bullatacin对HpG2细胞周期和凋亡及Fas蛋白表达的作用;RT-PCR检测Bullatacin对Fas基因表达的影响.结果:Bullatacin (100 gg/mL)能够明显抑制人肝癌细胞株HpG2的增殖,作用96 h其生长抑制率达66.3%;Bullataein将HpG2细胞周期抑制在G0/G1期,阻止其进入G2/M期,同时能诱导细胞凋亡;Bullatacin能够明显上调HpG2细胞表面Fas蛋白的表达(P=0.006),且作用优于顺铂(P=0.03);进一步研究发现,Bullatacin能够诱导Fas基因的表达.结论:Bullatacin能够诱导HpG2细胞凋亡,其可能机制是诱导Fas基因表达从而上调Fas蛋白.本研究以Bullatacin作为一种抗肿瘤药物的研发提供理论和实验依据.     

Enhancement of cytotoxic and pro-apoptotic effects of 2-aminophenoxazine-3-one on the rat hepatocellular carcinoma cell line dRLh-84, the human hepatocellular carcinoma cell line HepG2, and the rat normal hepatocellular cell line RLN-10 in combination with 2-deoxy-D-glucose

The cytotoxic and pro-apoptotic effects of a single dose of 2-aminophenoxazine-3-one (Phx-3) or 2-deoxyglucose (2-DG) or of a combined dose of Phx-3 and 2-DG were studied in the rat hepatocellular carcinoma cell line dRLh-84, the human hepatocellular carcinoma cell line HepG2 and the rat normal hepatocellular cell line RLN-10. The number of viable cells decreased in a dose-dependent manner, when dRLh-84, HepG2 or RLN-10 cells were treated with 2-DG (0.5-20?mM) or Phx-3 (1-50?μM) alone at 37?C for 48?h. When these cells were treated with 10?mM 2-DG and different concentrations of Phx-3, the number of viable cells decreased dose-dependently and in an additive manner for these agents. A single dose of 2 or 10?μM Phx-3 induced apoptotic morphology characterized by nuclear condensation and cell shrinkage in dRLh-84, HepG2 and RLN-10 cells, while a single dose of 10?mM 2-DG did not. When Phx-3 (2 or 10?μM) treatment was combined with 2-DG (10?mM) treatment in these three cell lines, the cells with apoptotic morphology increased extensively, which was confirmed by flow cytometric analysis. In addition, autophagic morphology characterized by cytosolic vacuole formation was significantly increased in the hepatocellular carcinoma cell lines dRLh-84 and HepG2 but not in the normal hepatocellular cell line RLN-10 after a single dose of Phx-3 or 2-DG or a combined dose of Phx-3 and 2-DG. Furthermore, when dRLh-84 and HepG2 cells were treated with Phx-3 alone or a combined dose of Phx-3 and 2-DG, depolarization of the mitochondria was extensive, but that of the normal cell line RLN-10 was not. These results may imply that the mechanism for the apoptosis of hepatocellular carcinoma cells caused by Phx-3 alone or a combined dose of Phx-3 and 2-DG differs from that of the normal cell line RLN-10. The present results demonstrate that Phx-3 alone may be beneficial for targeting liver cancer and that its anticancer activity may be enhanced by 2-DG. However, a combined dose of Phx-3 and 2-DG may exert adverse effects on normal liver cells, as evidenced by the cytotoxic and pro-apoptotic effects of the combined treatment in the rat normal hepatocellular cell line RLN-10.     

三氧化二砷诱导卵巢癌OVCAR-3细胞周期阻滞及凋亡

目的探讨As2O3对卵巢癌细胞增殖的抑制作用及对细胞周期和凋亡的影响.方法采用MTT法及集落形成实验测定细胞的增殖活力,通过细胞形态学观察细胞分裂及凋亡,应用流式细胞仪进行细胞周期解析、凋亡及bcl-2蛋白表达的检测.结果As2O3呈剂量依赖性抑制OVCAR-3细胞增殖,抑制50%细胞生长的药物浓度(IC50)为2μmol/L.0.5～5μmol/L的As2O3作用7天,集落抑制率均在40%以上(P＜0.01).2μmol/L与5μmol/L的As2O3作用12 h后,细胞周期出现了G2/M期阻滞及亚二倍体凋亡峰,随着作用时间的延长,凋亡细胞随G2/M期细胞减少而逐渐增多.细胞形态学观察可见分裂期细胞及凋亡细胞明显增加.0.5～5μmol/L的As2O3均能下调bcl-2蛋白的表达.结论As2O3能够抑制卵巢癌OVCAR-3细胞的增殖,诱导M期阻滞及细胞凋亡.     

Celecoxib inhibits MDR1 expression through COX-2-dependent mechanism in human hepatocellular carcinoma (HepG2) cell line

The role of COX-2 in the regulation of the expression of MDR1, a P-glycoprotein involved in hepatocellular carcinoma cell line, HepG2, was studied in the present investigation. Celecoxib, a selective inhibitor of COX-2, at 25 μM concentration increased the accumulation of doxorubicin in HepG2 cells and enhanced the sensitivity of the cells to doxorubicin by tenfold. The induction of MDR1 expression by PGE₂ and its downregulation by celecoxib or by COX-2 knockdown suggests that the enhanced sensitivity of HepG2 cells to doxorubicin by celecoxib is mediated by the downregulation of MDR1 expression, through COX-2-dependent mechanism. Further studies revealed the involvement of AP-1 in the celecoxib-induced downregulation of MDR1 expression. These experimental studies correlated well with in silico predictions and further suggested the inactivation of the signal transduction pathways involving ERK, JNK and p38. The present study thus demonstrates the usefulness of COX-2 intervention in overcoming the drug resistance in HepG2 cells.     

三氧化二砷对淋巴瘤细胞酪氨酸磷酸酶1基因的去甲基化作用

目的 探讨淋巴瘤细胞系T2和非霍奇金淋巴瘤(NHL)组织中酪氨酸磷酸酶1基因启动子区甲基化状态,三氧化二砷(As2O3)对T2细胞中SHP-1的去甲基化作用及对T2细胞生长增殖的生物学影响.方法 以不同浓度As2O3处理淋巴瘤细胞株T2,采用二苯基溴化四氮唑蓝(MTT)法检测T2细胞的生长变化,采用流式细胞术检测细胞凋亡率的变化,采用实时定量聚合酶链反应(FQ-PCR)和Western blot方法检测T2细胞SHP-1基因mRNA和蛋白表达及c-kit蛋白表达变化.采用甲基化特异性聚合酶链反应(MSP)检测T2细胞和32例NHL组织SHP-1基因启动子甲基化状态.结果 As2O3使T2细胞增殖受抑,凋亡增加,效应均具有时间和剂量依赖性.As2O3能逆转T2细胞SHP-1基因启动子甲基化,SHP-1基因恢复表达,同时c-kit蛋白磷酸化水平降低.SHP-1基因在对照组淋巴组织中呈完全性非甲基化状态,而在NHL组织中启动子甲基化出现率为87.5%(28/32).结论 在淋巴瘤细胞和NHL组织中,SHP-1基因启动子区域存在高度甲基化.As2O3能逆转T2细胞DNA异常甲基化,诱导SHP-1基因表达,并可能通过抑制c-kit受体及其信号传导路径的活化,抑制肿瘤细胞增殖.     

5-氮杂脱氧胞苷对肝癌细胞HepG2中抑癌基因FHIT表达的影响

背景与目的：肝癌细胞中由于FHIT基因过度甲基化而导致其表达降低。本实验应用甲基化酶抑制剂5-氮杂脱氧胞苷（5-Aza-2’deoxycytidine,5-Aza—dc）作用肝癌细胞株HepG2。观察用药后肝癌细胞中FHITmRNA和蛋白表达的变化,并观察5-Aza-dC对细胞增殖的影响。方法：以5-Aza—dC作用HepG2细胞．采用甲基化特异性聚合酶链反应（methylation—specific polymerase chain reaction,MSP）检测HepG2细胞中FHIT甲基化变化;采用RT-PCR检测FHITmRNA的表达：采用细胞免疫组织化学染色和Western blot方法检测FHIT蛋白表达,MTT法检测HepG2细胞增殖情况。结果：5-Aza—dC处理前,HepG2细胞FHIT基因呈甲基化状态,mRNA及蛋白表达缺失;经5-Aza—dC作用后,MSP显示HepG2细胞FHIT基因甲基化逆转：经1．0μmol／L、2．0μmol／L及4．0μmol／L的5-Aza—dC作用48h后,FHIT基因mRNA扩增出产物的吸光度值分别为0．80±0．32、1.41±0．54和1．51±0．61。Western blot检测产物积分灰度值分别为0．33±0．20、1．00±0．26和1．12±0．38：当药物浓度达到1．0μmol／L时出现了对HepG2细胞增殖的抑制作用。结论：5-Aza—dC能明显逆转HepG2细胞的FHIT基因异常甲基化,激活因高甲基化所致基因沉默的再转录,诱导该基因的表达;同时抑制细胞增殖。     

尼美舒利对肝癌细胞生长的影响

目的:探讨选择性环氧化酶-2(COX-2)抑制剂尼美舒利对肝癌细胞HepG2生长的影响.方法: MTs法检测对照组和实验组(尼美舒利25、50、100、200、400μmol/L)作用后HepG2细胞增殖变化;细胞侵袭小室法检测尼美舒利对HepG2迁移作用的影响.结果: 对肝癌细胞HepG2的增殖有抑制作用,且呈剂量依赖性,尼美舒利对HepG2细胞迁移有明显抑制作用.结论: 选择性环氧化酶2抑制剂尼美舒利可以抑制肝癌细胞的增殖和迁移.