凋亡抑制因子反义寡核苷酸对肝癌细胞生物学作用

目的 :探讨survivin反义寡核苷酸对人肝癌细胞凋亡、增殖、化疗药物敏感性的影响 .方法 :设计合成特异性survivin的反义寡核苷酸 (ASODN) .以高表达survivin基因的人肝癌细胞系 (SMMC 772 1 )为靶细胞、ASODN为阻断剂 ,Westernblot法检测细胞survivin表达情况 ,通过MTT试验观察ASODN对该细胞系的生长抑制作用 ,倒置显微镜观察细胞形态变化 ,流式细胞仪分析细胞增殖周期 ,MTT法检测survivin表达抑制前后细胞对紫杉醇敏感性影响 .结果 :ASODN明显抑制了SMMC 772 1肝癌细胞的生长 ,细胞分裂阻滞在分裂期的中期 ,并诱导细胞发生凋亡 ,而各对照组细胞生长良好 ;各A SODN药物组细胞凋亡指数明显高于各对照组 (P <0 .0 5 )且细胞增殖指数明显低于各对照组 (P <0 .0 5 ) ;紫杉醇药物组细胞IC5 0低于对照组 (1 6 .2± 2 .2 ) μg/Lvs (35 .5± 4 .3) μg/L ,P<0 .0 1 .结论 :survivinASODN能下调其蛋白表达 ,诱导人肝癌细胞凋亡 ,抑制细胞增殖 ,增加肝癌细胞对化疗药物的敏感性 .

c-myc反义寡核苷酸对表达FHIT基因的结肠癌细胞增殖及凋亡作用的影响

目的:观察脂质体介导的c-myc反义寡核苷酸对表达FHIT基因的结肠癌细胞增殖及凋亡作用的影响。方法:用脂质体法将重组FHIT基因的PRC/CMV质粒和空载体质转染到人结肠细胞株SW480,随后分别转染c-myc反义寡核苷酸。Western blot法检测细胞FHIT和c-myc的表达;MTT法检测细胞的增殖;AO/EB染色法和流式细胞分析技术检测细胞的凋亡。结果:转染FHIT基因后,SW480细胞有明显的FHIT蛋白表达而转染空载体的细胞未检测到FHIT蛋白表达。C-myc反义寡核苷酸转染后,对SW480细胞c-myc的表达有明显的抑制作用(P<0.01);对FHIT+/-SW480细胞的增殖均有明显的抑制作用(P<0.01),且对FHIT+SW480细胞的抑制作用明显强于FHIT-SW480细胞(P<0.05)。同时,c-myc反义寡核苷酸对FHIT+/-SW480细胞的凋亡均有明显的促进作用,对FHIT+SW480细胞凋亡的促进作用明显强于FHIT-SW480细胞(P<0.05)。结论:FHIT基因的表达和癌基因c-myc的表达抑制共同作用可以发挥较强的抗肿瘤细胞的效果,为多基因联合干预治疗肿瘤奠定了理论基础。

乙酰肝素酶反义硫代寡核苷酸对人肝癌细胞珠BEL-7402侵袭力的影响

目的:探讨乙酰肝素酶(Hpa)反义硫代寡脱氧核苷酸(AS-ODN)对人肝癌细胞BEL-7402侵袭力的影响。方法:设计合成互补于HpamRNA起始密码区的AS-ODN,以阳离子脂质体Oligofectami-neTMReagent包埋后转染BEL-7402细胞。采用RT-PCR和Western-blot检测不同浓度AS-ODN转染后细胞HpamRNA及蛋白表达的变化,以基底膜重建实验检测细胞侵袭力的变化。结果:0、100、200和300nmol/LAS-ODN转染组BEL-7402细胞HpamRNA的表达量分别为0·911、0·898、0·774和0·280;Hpa65×103蛋白表达量分别为0·923、0·875、0·834和0·237;Hpa50×103蛋白表达量分别为1·872、1·919、0·821和0·325。300nmol/LAS-ODN转染后BEL-7402细胞HpamRNA和蛋白的表达明显下降;0、100、200和300nmol/LAS-ODN转染组侵袭细胞数分别为137±15、141±13、134±18和49±7,300nmol/L,AS-ODN转染组侵袭细胞数较正常对照组明显减少,P=0·000。结论:一定浓度的HpaAS-ODN可通过下调HpamR-NA和蛋白的表达抑制肝癌细胞的侵袭力。

AT_1R反义寡核苷酸对心肌细胞生长影响的实验研究

目的 :探讨 AT1 R反义寡核苷酸 (AT1 R- AS- ODNs)对正常大鼠心肌细胞生长的影响。方法 :脂质体介导AT1 R- AS- ODNs转染体外培养的 SD乳鼠心肌细胞 ,Western印迹及原位杂交检测细胞内 AT1 R蛋白及 m RNA表达 ;流式细胞仪检测细胞内 DNA合成 ;免疫沉淀法检测 c- Jun氨基末端活化蛋白激酶 (JNKs)活性 ;免疫细胞化学检测核转录因子 c- Jun蛋白表达水平。结果 :在脂质体介导下 ,AT1 R- AS- ODNs成功转染心肌细胞 ;转染 2 4 h后 ,AT1 R蛋白及 m RNA表达水平分别被下调 6 0 .7%、5 1.2 % (P<0 .0 5 ) ;但 AT1 R- AS- ODNs转染对细胞内 JNKs活性、c- Jun蛋白表达、DNA合成无显著影响。结论 :AT1 R- AS- ODNs能有效抑制正常大鼠心肌细胞 AT1 R表达 ,但不改变细胞内 AT1 R偶联的信号转导过程 ,也不影响细胞的正常生长。

Caspase-3 mRNA反义核苷酸对γ-射线诱导的HL-60细胞凋亡的影响(英文)

目的观察caspase-3 mRNA反义寡核苷酸(ASODN)对HL-60细胞凋亡的抑制作用,筛选有效ASODN。方法用脂质体介导法将针对caspase-3 mRNA不同序列的4条ASODN导入HL-60细胞中,γ-射线照射。应用电泳法检测DNA梯状条带;Hoechst 33258-碘化丙啶染色,荧光显微镜分析凋亡细胞百分率;流式细胞术进行细胞凋亡定量。结果以caspase-3 mRNA5’非编码区(-62至-46位)与编码起始区(-1至16位)ASODN转染,当转染终浓度≥3μmol/L时,DNA电泳梯状条带消失,流式细胞术亦未见明显的亚二倍体峰;荧光染色分析,凋亡细胞百分率比未转染对照组和错配寡核苷酸对照组显著降低(P<0.01),且随转染终浓度的增加,凋亡抑制率显著增加。另外,5’非编码区ASODN的抑制作用显著强于编码起始区ASODN(P<0.05)。结论caspase-3 mRNA ASODN可抑制γ-射线诱导的HL-60细胞凋亡,作用有序列特异性及剂量依赖性。该结果为细胞过度凋亡相关疾病的基因治疗提供了实验基础。

内皮素A型受体反义基因减弱内皮细胞条件培养液刺激的血管平滑肌细胞增殖

目的 :研究内皮素 (Endothelin,ET) A型受体 (ETA)反义寡核苷酸 (Oligonucleotide,OGN)对内皮细胞 (En-dothelial cells,ECs)条件培养液 (ECCM)刺激的人大隐静脉 (hum an saphenous vein,HSV)血管平滑肌细胞 (Vas-cular sm ooth m uscle cells,VSMCs)增殖的影响。方法 :分离、培养 HSV的 ECs和 VSMCs。收集 ECCM,分别用正常的 DMEM(Dulbecco's Mod Ified Eagle medium )、含 ET- 1的 DMEM、ECCM和加入 Phophoramidon(ET- 1转化酶抑制剂 )的 ECCM培养 VSMCs。以放射免疫法测定 ECs培养基中 ET- 1水平 ;分别采用放射受体结合分析法和四唑盐 (MTT)比色实验检测 ETA 蛋白表达和细胞的增殖。结果 :ETA- OGN(15 μm ol/L)以时间依赖的方式抑制ETA 表达。 12～ 4 8h HSV的 ECs培养上清液 ET- 1水平呈现上升的趋势 ,4 8h达到高峰。 ECCM显著刺激 VSMCs增殖 (P<0 .0 1) ,ET- 1能够模拟 ECCM的促增殖作用。在 ECCM中加入 Phosphorimidon,不能使 ECCM的促增殖效应消失 (P<0 .0 5 )。ETA- OGN(15 μm ol/L)对无血清正常的 DMEM培养基培养的 VSMCs增殖没有显著影响 ,但显著抑制 ECCM和 ET- 1的促增殖作用 (P<0 .0 1vs ECCM)。结论 :ECs通过分泌包括 ET- 1在内的多种因子促进 VSMCs增殖 ,其中 ET- 1发挥主要的作用。

肾癌相关新基因GYLZ-RCC18反义寡核苷酸转染对肾癌细胞系GRC-1的作用

目的 探讨肾癌相关新基因GYLZ RCC18在肾癌发生发展中的作用机制。 方法 采用逆转录PCR方法检测GYLZ RCC18在肾癌和正常肾细胞系中的表达 ,将第一编码区起始处的2 0个碱基的反义和正义寡核苷酸导入肾癌细胞系GRC 1,连续观察对肾癌细胞生长、增殖凋亡、死亡率、形态学的影响。 结果 GYLZ RCC18在肾癌细胞系中表达明显高于正常肾细胞系 ;导入GYLZ RCC18反义寡核苷酸后 ,GRC 1细胞的核分裂可明显减少 ,导致GRC 1细胞死亡 ,抑制癌细胞的生长和增殖活性 ,并可特异诱导GRC 1连续凋亡。 结论 GYLZ RCC18是肾癌发生发展密切相关的癌基因 ,其表达可促进肾癌细胞的生长和增殖 ,并通过抗癌细胞死亡和凋亡机制对癌细胞起保护作用。

TGFα反义寡聚核苷酸抑制大肠癌细胞株的增殖

研究ＴＧＦａ反义家聚核苷酸抑制大肠癌ＨＲ８３４８细胞株恶性增殖的作用。方法采用２３个碱基组成的ＴＧＦα反义寡聚核苷酸和２１个碱基组成的对照寡聚核苷酸作用于人大肠癌ＨＲ８３４８细胞，通过细胞生长抑制实验、Ｈ－ＴｄＲ掺入、ｍＲＮＡ斑点杂交及细胞周期分析以确定其对大肠癌细胞的增殖抑制作用和作用环节。结果ＴＧＦα反义寡聚核苷酸均能抑制ＨＲ８３４８细胞的增殖、ＤＮＡ合成和ＴＧＦαｍＲＮＡ的表达，并能有效地延缓ＨＲ８３４８细胞由Ｇ０／Ｇ１期向Ｓ期的转化，延缓了细胞的分裂增殖。结论ＴＧＦα反义寡聚核苷酸能有效地抑制ＨＲ８３４８细胞的恶性增殖。

联载多西紫杉醇和反义寡聚脱氧核糖核苷酸脂质体的制备工艺

目的研究联载多西紫杉醇(Doc)、反义寡聚脱氧核糖核苷酸(ODN)脂质体的制备工艺。方法采用乙醇注入法制备脂质体,以包封率和粒径为主要评价指标来考察脂质体的制备工艺,采用中心组合实验进一步优化处方;用激光粒度仪测量脂质体的粒径;采用HPLC法测定脂质体中Doc的包封率,将ODN用FITC标记,用荧光分光光度计测量荧光强度以测量ODN的包封率。结果最佳处方为磷脂-总胆固醇(3∶1),DOC药脂比为1∶50,胆固醇(Chol)-阳离子胆固醇(DC-Chol)为2.4∶1,Chol-PEG2000含量8%,Doc包封率为50%～60%,ODN包封率为60%～70%。结论用乙醇注入法成功地制备了双载药脂质体。

Efficacy of Antisense Oligodeoxynucleotide of Neuropeptide Y Y5 Receptor on Treating of Hyperleptinemia by Intraventricular Administration in Diet-induced Obese Rats

Objective: To study the efficacy of antisense oligonucleotide of neuropeptide Y (NPY) Y5 receptor on treating hyperleptinemia by intracerebral ventricular administration in diet-induced obese rats.Methods: The obese rats were prepared by feeding a high-nutritive diet for 7 weeks. The lateral ventricle of obese rats was cannulated. Either 10 μl of different neuropeptide Y Y5 receptor oligodeoxynucleotide, including antisense, sense and missense oligodeoxynucleotide (5 g/L) or 10 μl saline was administered into the ventricle through cannula three times per day in every rat. Two days later the rats were slaughtered .The weights of both retroperitoneal and epididymal adipose tissues were measured, and the serum insulin and leptin were detected by radioimmunoassay method and the murine leptin ELISA kit respectively. Results: ①The level of serum was significantly higher in experimental rats than that in normal rats. Similarly, the level of serum insulin and the weights of both retroperitoneal and epididymal adipose tissues were increased in experimental rats. ②After the diet-induced obese rats were intraventricularly administered with NPY Y5 receptor antisense oligodeoxynucleotide, the levels of serum leptin and insulin were significantly decreased and combined with the reduction of weight in retroperitoneal adipose tissue. There was, however, no significant difference in the weight of epidymal adipose tissue between pre-treated and post-treated duration. ③There was significant positive correlation among the level of serum leptin, the level of serum insulin and the weight of retroperritoneal adipose tissue in diet-induced obese rats. Conclusion: Intracerebral ventricular administration of antisense oligodeoxynucleotide of neuropeptide Y Y5 receptor may alleviate hyperleptinemia in diet-induced obese rats and decrease the weight of retroperitoneal adipose tissue and the level of serum insulin.

Effects of Ghrelin Antisense Inhibition on VEGF and Its Receptor Flt-1 mRNA Expression

[ Objective] This study was to investigate the effect of VEGF and its receptor Fit-1 mRNA expression in Mongolia sheep umbilical vein endothelial cells by ghrelin antisense inhibition. [ Method] Experiments were divided into 4 groups： group Ⅰ （blank control group） ; group Ⅱ （liposome group） ; group Ⅲ （SCON group： 20 μmol/L sense oligonucleotide） ; group Ⅳ （ASCON： 20 μmol/L antisense oligonucleotide）. VEGF and its receptor Fit-1 mRNA expression changes were detected by using real-time fluorescence quantitative detection after 24, 36 and 48 h. [ Result] The expression of VEGF mRNA in group Ⅰ, group Ⅱ were insignificantly different at higher expression levels, and did not change significantly with the time; the expression of VEGF mRNA in group Ⅲ assumed a slight decrease, but there were no significant differences between group I and group Ⅱ （P 〉0.05）, the expression of VEGF mRNA in group Ⅳ（antisense oligonucleotide group ） decreased significantly （P 〈 0.05） ; the expression of VEGF receptor FLT-1 mRNA was similar to that of VEGF. [ Conclusion] Antisense inhibition ghrelin has a downward effect to the expression of VEGF and its receptor Fit-1 the mRNA.

Antisense oligonucleotide targeting midkine suppresses in vivo angiogenesis

AIM: To evaluate the effect of antisense oligonucleotide targeting midkine (MK-AS) on angiogenesis in chick chorioallantoic membrane (CAM) and in situ human hepatocellular carcinoma (HCC). METHODS: An in situ human hepatocellular carcinoma (HCC) model and CAM assay were used in this experiment. The effect of MK-AS on angiogenesis was evaluated by cell proliferation assay and hematoxylin- eosin (HE) staining. RESULTS: MK-AS significantly inhibited human umbilical vein endothelial cells (HUVEC) and in situ human HCC growth. At the same time, MK-AS suppressed the angiogenesis both in human hepatocellular carcinoma cell line (HEPG2)-induced CAM and in situ human HCC tissues. CONCLUSION: MK-AS is an effective antiangiogenesis agent in vivo.

Enhanced therapeutic effects of combined chemotherapeutic drugs and midkine antisense oligonucleotides for hepatocellular carcinoma

AIM: To evaluate the effect of combined antisense oligonucleotides targeting midkine (MK-AS) and chemotherapeutic drugs [cisplatin(DDP), 5-fluorouracil (5-FU) and adriamycin (ADM)] on inhibition of HepG2 cell proliferation, and to analyze the efficacy of MK-AS used in combined ADM in in situ human hepatocellular carcinoma (HCC) model. METHODS: HepG2 cells were treated with MK-AS and/or chemotherapeutic drugs mediated by Lipofectin, and cell growth activity was determined by MTS assay. An in situ HCC model was used in this experiment. MK- AS, ADM and MK-AS + ADM were given intravenously for 20 d, respectively. The animal body weight and their tumor weight were measured to assess the effect of the combined therapy in vivo. RESULTS: Combined treatment with MK-AS reduced the IC50 of DDP, 5-FU and ADM in HepG2 cells. MK-AS significantly increased the inhibition rate of DDP, 5-FU and ADM. Additionally, synergism (Q 1.15) occurred at a lower concentration of ADM, 5-FU and DDP with combined MK-AS. Combined treatment with MK-AS and ADM resulted in the more growth inhibition on in situ human HCC model compared with treatment with chemotherapeutic drugs alone. CONCLUSION: MK-AS increases the chemosensitivity in HepG2 cells and in situ human HCC model, and thecombination of MK-AS and ADM has a much better in vitro and in vivo synergism.

The Role of β3-adrenergic Receptor Gene in Neuropeptide Y Y5Receptor Antisense Gene Therapy of Diet-induced Obese Rats

Objective: To study the role of β3-adrenergic receptor gene in neuropeptide Y(NPY) Y5 receptor antisense gene therapy of diet-induced obese rats.Methods: The diet-induced obese rats were prepared by feeding a high-nutrition diet. Lateral ventricular was cannulated in obese rats which then received an intraventricular injection of either 5 μg/μl NPY Y5 receptor antisense or 10 μl missense oligodeoxynucleotide or saline of 10 μl respectively in every rat. When the rats were killed, the wet weight of abdominal adipose tissue, the level of serum lipid and lipoprotein were measured. Total RNA from the retroperitoneal adipose tissue was extracted and the level of β3-adrenergic receptor gene mRNA expression was evaluated by RT-PCR.Results: ①The wet weight of abdominal adipose tissue, the levels of serum lipids were greatly higher in diet-induced obese rats than those in normal rats. However, there were much lower β3-adrenergic receptor gene mRNA expression levels in retroperitoneal adipose tissue in diet-induced obese rats as compared with those in normal rats. ②After the diet-induced obese rats were intraventricularly administered with NPY Y5 receptor antisense oligodeoxynucleotide, the levels of β3-adrenergic receptor gene mRNA expression in retroperitoneal adipose tissue of diet-induced obese rats were strikingly up-regulated, whereas the wet weight of abdominal adipose tissue, the levels of serum lipids were markedly reduced.Conclusion: Intraventricular administration of antisense oligodeoxynucleotide to NPY Y5 receptor could significantly reduce the abdominal adipose tissue and the levels of serum lipids in diet-induced obese rats by up-regulating the level of β3-adrenergic receptor gene mRNA expression in retroperitoneal adipose tissue.

In vitro and in vivo suppression of hepatocellular carcinoma growth by midkine-antisense oligonucleotide-loaded nanoparticles

AIM:To synthesize antisense oligonucleotides (ASODNs) of midkine (MK), package the ASODNs with nanoparticles, and to inhibit hepatocellular carcinoma (HCC) growth using these nanoparticles.METHODS: HepG2 cell proliferation was analyzed in vitro using the 3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt assay. The in vivo activity of nanoparticles delivering the MK-ASODNs was analyzed by histopathological and immunohistochemical staining and quantitative real time polymerase chain reaction (PCR). RESULTS: The in vitro proliferation of HepG2 cells was signif icantly inhibited by the nanoparticles packaged with MK-ASODNs (NANO-ASODNs). Furthermore, the NANO- ASODNs signif icantly inhibited the growth of HCC in the mouse model. CONCLUSION: NANO-ASODNs can significantly suppress the growth of HCC in vitro and in vivo.

Suppression of intracranial glioma tumorigenesis with vascular endothelial growth factor antisense oligonucleotide in rats

Objective: To observe the inhibition of intracranial glioma tumorigenesis by vascular endothelial growth factor (VEGF) antisense oligodeoxynucleotide (ODN) in rats. Methods: Totally 20 μ1 Hank's liquid containing 1×106 C6 glioma cells was seeded into rat right caudate putaraen in high-flow microinfusion with stereotactic technique. VEGF antisense ODN was simultaneously used with glioma cell. Each rat of the treated group Ⅰ and the treated group Ⅱ was treated with 1 000 μmol/L VEGF antisense ODN. Each rat of the treated group Ⅲ and the treated group Ⅳ was treated with 2 000 μmol/L VEGF antisense ODN. The experimental periods of the treated group Ⅰ , the treated group Ⅲ and the control group Ⅰ were 2 weeks, those of the treated group Ⅱ , the treated group Ⅳ and the control group Ⅱ were 3 weeks. Before sacrifice, MRI was performed on each rat. Tumor magnitude and pathologic examination were detected after samples were dissected. Results: The survival state of all treated rats was better, and that of the control rats was in severe danger. The tumor volumes of the treated group Ⅰ and the treated group Ⅱ were remarkably lessened. Tumor tissue could not be found macroscopically in the brain samples of the treated group Ⅲ and the treated group Ⅳ, but tumor nest could be found with microscopy. Tumors of the treated group I and the treated group Ⅱ had weak expressions of VEGF mRNA and VEGF, while normal brains and the samples of the treated group Ⅲ and the treated group Ⅳ had negative expressions, but tumors of the control groups had strong expressions. Conclusion: VEGF antisense ODN used early in situ can suppress angiogenesis and growth of rat intracranial glioma to retard tumorigenesis.

Role of Smad7 in inflammatory bowel diseases

Crohn's disease and ulcerative colitis,the major forms of inflammatory bowel diseases(IBD) in man,are complex diseases in which genetic and environmental factors interact to promote an excessive mucosal immune response directed against normal components of the bacterial microflora.There is also evidence that the pathologic process is due to defects in counterregulatory mechanisms,such as those involving the immunosuppressive cytokine transforming growth factor(TGF)-1.Indeed,studies in human IBD tissues and murine models of colitis have documented a disruption of TGF-1 signalling marked by a block in the phosphorylation of Smad3,a signalling molecule associated with the activated TGF-receptor,due to up-regulation of Smad7,an intracellular inhibitor of Smad3 phosphorylation.Knock-down of Smad7 with a specific antisense oligonucleotide restores TGF-1/Smad3 signalling,thus resulting in a marked suppression of inflammatory cytokine production and attenuation of murine colitis.These findings together with the demonstration that Smad7 antisense oligonucleotide is not toxic when administered in mice have paved the way for the development of a Smad7 antisense oligonucleotidebased pharmaceutical compound that is now ready to enter the clinics.In this article we review the available data supporting the pathogenic role of Smad7 in IBD and discuss whether and how Smad7 antisense therapy could help dampen the ongoing inflammation in IBD.

Effect of transfected angiotensinⅡ receptor anti-sense nucleotide on the growth of cardiomyocytes in vitro

Objective:To evaluatetheeffectof transfectingangiotensinⅡreceptor(AT1)anti-sensenucleotide(AT1A)on theexpressionof subtypesof AngiotensinⅡ(AngⅡ)receptormRNA,andsynthesesof proteinand nucleicacidincardiomyocytes.Methods:AT1cDNAsequence(476bp)wasclonedwithRT-PCRandinsertedinto PcDNA3.1(5.4kb)anti-senselyto constructan intactplasmidcontainingAT 1 A(PAT 1 A).It was transfectedintothe culturedcardiomyocytes,whichwasidentifiedwithRT-PCRandWesternblot.Synthesesof proteinandnucleicacid weredeterminedwith 3 H-Leuand 3 H-TdRincorporation,mRNAexpressionsof AT 1 andAT 2 wereobservedwith RT-PCR.Transfectedandnontransfectedcardiomyocyteswerecomparedafterstimulatedfor24h by AngII1×10 -7 mol/L.Results:WeconstructedPAT 1 A successfully.AT 1 mRNAandproteinwereexpressedsignificantlylessin transfectedcardiomyocytesthanthatin thecontrol(P<0.01).AT 1 mRNAexpressionwas markedlydecreased,and AT 2 mRNAobviouslyincreased(P<0.01);butno apparentdifferencewas foundin 3 H-Leucine( 3 H-Leu)and 3 H-Thymidine( 3 H-TdR)incorporationbetweenthetransfectedandnontransfectedcardiomyocytesafterstimulated for24h of AngⅡ10 -7 mol/L(P>0.05).Conclusion:AfterblockedwithAT 1 A,expressionof AT 1 mRNAincultured cardiomyocyteswas markedlysuppressed,whileAT 2 mRNAwas up-regulatedat thesametime.Thisfactsuggests thatsynthesesof bothproteinand nucleicacidin cardiomycytesmediatedwithAng II couldnot be effectively interruptedsimplywithAT1Ablocking.

Inhibitory effects of transfected Bcl-XL antisense oligodeoxynucleotide to proliferation and growth in esophageal carcinoma cell line and human esophageal carcinoma xenograft of nude mice

Objective： The aim of our study was to investigate the biological effects of Bcl-XL antisense oligodeoxynucleotide （ASODN） transfected into cultured esophageal carcinoma cells and human esophageal carcinoma xenograft in nude mice. Methods： Cationic liposome-mediated ASODN was used to transfect esophageal carcinoma cells. RT-PCR, Western blot, MTT assay, flow cytometry, and in situ apoptosis cells detection （TUNEL detection） were used to systematically study the biological effects of transfected cells both in vitro and in vivo. Results： In this study, the results showed that the proliferation of esophageal carcinoma cells in ASODN group decreased significantly when compared with the control group （P 〈 0.05）, at 57.3% Bcl-XL mRNA inhibitory rate, and a significant decreasing of Bcl-XL protein expression, at the apoptosis rates of （31.1 ＋ 5.8）% and 35.0% by flow cytometry and TUNEL assay respectively （P 〈 0.01, when compared with control groups）. It also showed that the growth of human esophageal carcinoma in nude mice of ASODN group was significantly inhibited （P 〈 0.05）, together with a significant decreased expression level of Bcl-XL mRNA and protein, and an induced tumor cell apoptosis in nude mice. Conclusion： Our result indicates BcI-XL ASODN can effectively inhibit the proliferation of esophageal carcinoma cells in vitro and tumor growth in vivo. The suppression of Bcl-XL expression by ASODN may offer both a therapeutic approach and an important theoretic foundation for gene therapy against esophageal carcinoma.