Tumor suppressor in lung cancer-1 (TSLC1) mediated by dual-regulated oncolytic adenovirus exerts specific antitumor actions in a mouse model

Aim:

The tumor suppressor in lung cancer-1 (TSLC1) is a candidate tumor suppressor of lung cancer, and frequently inactivated in primary non-small cell lung cancer (NSCLC). In this study, we investigated the effects of TSLC1 mediated by a dual-regulated oncolytic adenovirus on lung cancer, and the mechanisms underlying the antitumor actions.

Methods:

The recombinant virus Ad·sp-E1A_(Δ24)-TSLC1 was constructed by inserting the TSLC1 gene into the dual-regulated Ad·sp-E1A_(Δ24) vector, which contained the survivin promoter and a 24 bp deletion within E1A. The antitumor effects of Ad·sp-E1A_(Δ24)-TSLC1 were evaluated in NCI-H460, A549, and H1299 lung cancer cell lines and the normal fibroblast cell line MRC-5, as well as in A549 xenograft model in nude mice. Cell viability was assessed using MTT assay. The expression of TSLC1 and activation of the caspase signaling pathway were detected by Western blot analyses. The tumor tissues from the xenograft models were examined using H&E staining, IHC, TUNEL, and TEM analyses.

Results:

Infection of A549 lung cancer cells with Ad·sp-E1A_(Δ24)-TSLC1 induced high level expression of TSLC1. Furthermore, the Ad·sp-E1A_(Δ24)-TSLC1 virus dose-dependently suppressed the viability of NCI-H460, A549, and H1299 lung cancer cells, and did not affect MRC-5 normal fibroblast cells. Infection of NCI-H460, A549, and H1299 lung cancer cells with Ad·sp-E1A_(Δ24)-TSLC1 induced apoptosis, and increased activation of caspase-8, caspase-3 and PARP. In A549 xenograft model in nude mice, intratumoral injection of Ad·sp-E1A_(Δ24)-TSLC1 significantly suppressed the tumor volume, and increased the survival rate (from less than 15% to 87.5% at d 60). Histological studies showed that injection of Ad·sp-E1A_(Δ24)-TSLC1 caused tumor cell apoptosis and virus particle propagation in tumor tissues.

Conclusion:

The oncolytic adenovirus Ad·sp-E1A_(Δ24)-TSLC1 exhibits specific antitumor effects, and is a promising agent for the treatment of lung cancer.     

Effect of trichostatin A and paclitaxel on the proliferation and apoptosis of lung adenocarcinoma cells

Background Histone deacetylase inhibitors can regulate gene expression through modulation of the degree of acetylation of histone and non-histone,thus affecting cell proliferation,survival and chemosen...     

靶向生存素siRNA抑制人喉鳞癌裸鼠动物模型的实验研究

目的 研究靶向生存素siRNA对喉鳞癌动物模型的抑制作用,探讨其可能的抑瘤机制.方法 建立人喉鳞癌裸鼠移植瘤模型,将33只裸鼠随机分为3组,对照组瘤体内注射PBS液,空载体组瘤体内注射空慢病毒载体(Ad-control),治疗组应用生存素siRNA (Ad-survivin)进行瘤内注射治疗,观察各组肿瘤生长情况,光镜及透射电镜观察肿瘤超微结构变化,TUNEL法检测细胞凋亡,免疫组化法检测Bcl-2蛋白的表达情况.结果 治疗组体内注射生存素siRNA(100 ng·mL-1·d-1),裸鼠肿瘤生长受到明显抑制,与对照组及空载体组相比,差异有统计学意义(P＜0.05).光镜、电镜下观察到治疗组肿瘤组织发生坏死和凋亡改变,Tunel证实治疗组肿瘤细胞凋亡数目明显增多,与对照组及空慢病毒组相比,差异有统计学意义(P＜0.05),免疫组化示治疗组Bcl-2表达下降.结论 生存素siRNA抑制喉鳞癌裸鼠生长,其作用机制可能与抑制Bcl-2表达诱导肿瘤细胞凋亡相关.     

卵巢上皮癌旁不同距离组织中survivin及MMP-2/TIMP-2的表达

目的通过对卵巢上皮癌旁不同距离组织的survivin及基质金属蛋白酶2(MMP-2)/基质金属蛋白酶组织抑制剂2(TIMP-2)的表达量检测,探讨获取癌旁正常卵巢组织的安全界限。方法采用RT-PCR检测20例正常卵巢组织,32例卵巢上皮癌组织及其癌旁0.5cm、1.0cm、1.5cm处组织的survivin及MMP-2/TIMP-2的表达量。结果癌旁0.5cm处的卵巢组织survivin及MMP-2/TIMP-2表达量明显高于正常组织(P<0.01),略低于上皮癌组织但无统计学差异(P>0.05);癌旁0.5～1.0cm处的卵巢组织survivin及MMP-2/TIMP-2的表达量明显低于卵巢癌组织(P<0.01),略高于正常卵巢组织但无统计学差异(P>0.05);癌旁1.0～1.5cm处的卵巢组织survivin及MMP-2/TIMP-2的表达量和正常卵巢组织差异无统计学意义(P>0.05);两种病理类型的卵巢上皮癌旁组织survivin及MMP-2/TIMP-2表达量差异无统计学意义(P>0.05)。结论癌旁0.5cm以内的组织可能是肿瘤隐匿转移的危险区域,癌旁1.0cm可能是获取正常卵巢组织的最小安全距离。     

骨髓间充质干细胞抗辐射基因Survivin和HO-1的表达

本研究探讨人骨髓间充质干细胞抗辐射基因survivin和HO-1的表达。采用Fircoll密度梯度离心法自人骨髓中分离MSC并进行纯化和扩增,通过流式细胞术检测其表面标志并进行鉴定;用地塞米松、胰岛素、3-异丁基-1-甲基黄嘌呤(IBMX)、吲哚美辛等定向诱导BM-MSC分化为脂肪细胞;RT-PCR检测MSC中survivin和HO-1基因表达。结果表明:体外分离培养的MSC中CD34、HLA-DR表达为阴性,CD71、CD44表达为阳性,并且可诱导为脂肪细胞。RT-PCR检测MSC显示有survivin和HO-1抗辐射基因表达。结论 :MSC具有较低的辐射敏感性,这可能与抗辐射基因survivin和HO-1表达相关。     

支气管灌洗液Survivin基因检测在肺癌诊断中的价值

目的 探讨支气管灌洗液(BLF)标本Survivin m RNA检测在肺癌尤其是周围型肺癌诊断中的可行性及应用价值.方法 应用逆转录-聚合酶链反应(RT-PCR)方法,分别检测124例肺癌、103例支气管肺部良性疾病患者BLF标本Survivin mRNA表达情况,并与BLF标本细胞病理学等结果相比较.结果 BLF标本Survivin mRNA表达阳性率肺癌患者45.2％(56/124),显著高于支气管肺部良性疾病患者18.4％(19/103),差异有统计学意义(x2=18.150,P=0.000).BLF标本Survivin mRNA相对表达量与肺癌病理类型、分化程度均相关(均P ＜0.05);而与肺癌患者的性别、年龄、抽烟史、肿瘤的部位、大小、淋巴结转移、远处转移、临床分期及中心型肺癌患者BLF方式之间均无明显相关(均P＞0.05).BLF标本检测Survivin mRNA诊断肺癌的敏感性、特异性分别为45.2％(56/124)和81.6％(84/103),诊断周围型肺癌的敏感性、特异性分别为35.9％(14/39)和85.4％(88/103).BLF标本检测Survivin mRNA表达联合细胞病理学检查阳性率为57.5％(65/113),显著高于单独检测Survivin mRNA阳性率43.4％(49/113)或细胞病理学阳性率23.9％(27/113),差异均有统计学意义(x2=4.531,26.472,P=0.033,0.000).结论 BLF标本Survivin mRNA可能成为肺癌诊断有价值的肿瘤标志物之一,为肺癌尤其是周围型肺癌早期诊断提供帮助,BLF标本获取方式不影响Survivin mRNA表达检测结果.     

Survivin、caspase3蛋白的表达与颊鳞癌发病机制研究

目的研究生存素survivin和半光酰天冬氨酸特异性蛋白酶3(cysteiny aspirate specific protease-3,caspase-3)在颊鳞状细胞癌中的表达情况,探讨凋亡抑制蛋白survivin和caspase-3在颊鳞状细胞癌中的表达特点及其相互关系,以及两者在颊鳞状细胞癌发病机制中的可能作用。方法采用免疫组织化学SP法检测随机抽取的8例颊部正常组织、8例癌前病变、16例颊鳞癌石蜡切片中survivin和caspase 3的表达,并统计分析与颊鳞癌病理分级与survivin和caspase-3表达水平之间的关系。结果 16例口腔颊癌中survivin表达阳性14例,阳性率为87.5%,Caspase 3阳性表达5例,阳性率为31.3%,颊部癌前病变组中survivin表达阳性7例,阳性率为87.5%,Caspase 3阳性表达7例,阳性率为87.5%,8例正常组织中survivin 0表达,Caspase 3完全表达,survivin、Caspase 3在颊癌和颊部癌前病变中表达的差异有统计学意义(P<0.05)。结论口腔颊癌组织中survivin和Caspase 3的表达具有相关性,可以认为survivin过度表达抑制Caspase 3的活化,抑制细胞的凋亡是口腔颊癌发病的机制之一,同时可以将二者作为预后和抗肿瘤治疗疗效判定的指标之一。     

Decreased immunoexpression of survivin could be a potential marker in human non‐alcoholic fatty liver disease progression?

Background/aim: Regulation of apoptosis in non‐alcoholic fatty liver disease (NAFLD) has been a theme of growing debate. Although no other study assessed the role of survivin in NAFLD, its expression has been reported in hepatic carcinogenesis because of other aetiological factors with relevant discrepancies. The aim of this study was to assess the pattern of survivin immunoexpression by tissue microarray along the whole spectrum of NAFLD, including non‐alcoholic steatohepatitis (NASH)‐related hepatocelular carcinoma (HCC). Methods: Liver biopsies from 56 patients with NAFLD were evaluated: 18 with steatosis, 21 non‐cirrhotic NASH, 10 NASH‐related cirrhosis, seven NASH‐related HCC, as compared with 71 HCC related to other causes and with 12 normal livers. Results: Survivin immunoexpression in NAFLD was restricted to cytoplasm and was found to be progressively lower in advanced stages, including cirrhosis and HCC: steatosis vs NASH‐related cirrhosis (P=0.0243); steatosis vs NASH‐related HCC (P=0.0010); NASH vs NASH‐related cirrhosis (P=0.0318); and NASH vs NASH‐related HCC (P=0.0007), thus suggesting a deregulation of apoptosis from NAFLD towards HCC. Interestingly, survivin immunoreactivity in NASH‐related HCC was also found to be significantly lower than in HCC related to other causes (P<0.05). Remarkably, nuclear staining for survivin was not detected in any case of NAFLD, contrasting to its presence in all other cases of HCC. Conclusions: Survivin immunoexpression in NASH‐related HCC is herein originally found substantially different than in HCC related to other causes, thus requiring further studies to elucidate the role of survivin in human NAFLD progression.