舌鳞癌层粘连蛋白的表达及意义

采用免疫组织化学技术,研究了正常、异常增生组织、舌癌及舌癌淋巴结转移灶中层粘连蛋白的表达、分布及意义。结果发现：正常舌粘膜、轻度异常增生组织的基底膜处层粘连蛋白呈连续线状分布,中度和重度异常增生组织基底膜层粘连蛋白的分布有局部断裂。舌癌中层粘连蛋白的分布呈多种形式,分化好的舌癌层粘连蛋白呈线状,但明显不连续,分化差的舌癌中的层粘连蛋白常呈碎片状,而舌癌的淋巴结转移灶中层粘连蛋白的分布与原发灶相似。统计分析表明,层粘连蛋白的表达与舌癌的分化程度相关,而与舌癌的转移无关。观察结果表明：舌癌分化程度愈低,层粘连蛋白的缺损愈严重,层粘连蛋白的表达、分布特点可以作为判断癌恶性程度的指标,但不能判断舌癌的转移     

Bmi1 在肝癌组织中的表达及其与细胞增殖和凋亡的关系

目的:探讨Bmi1在肝细胞肝癌(HCC)中的表达及与增殖和凋亡的关系。方法:收集HCC标本54例及相应的癌旁组织,10例正常肝组织标本,采用免疫组织化学EnVision二步法显示Bmi1的表达并结合增殖与凋亡特征进行分析。结果:在肝癌组织、癌旁组织细胞中Bmi1表达定位于胞核中,阳性表达率分别为79.6%(43/54),31.2%(17/54),10例正常肝组织未见表达,3组差异有统计学意义(P<0.005)。HCC中Bmi1的高表达与年龄、性别、肿瘤大小、肿瘤数目、临床TNM分期、是否有肝硬化及是否有HBsAg感染无显著相关(P>0.05),但与组织学分级有关,高、中分化组Bmi1表达率显著高于低分化组(P<0.05)。肝癌Bmi1阳性组增殖指数(PI)(50.3±21.4)%显著高于阴性表达组(17.3±7.1)%(P<0.05),凋亡指数(AI)无明显差异(P>0.05)。结论:Bmi1在HCC中高表达,其表达增高可能与HCC的进展相关。     

Bmi1在肝癌组织中的表达及其与细胞增殖和凋亡的关系

目的：探讨Bmi1在肝细胞肝癌（HCC）中的表达及与增殖和凋亡的关系。方法：收集HCC标本54例及相应的癌旁组织,10例正常肝组织标本,采用免疫组织化学EnVision二步法显示Bmi1的表达并结合增殖与凋亡特征进行分析。结果：在肝癌组织、癌旁组织细胞中Bmi1表达定位于胞核中,阳性表达率分别为79.6%（43/54）,31.2%（17/54）,10例正常肝组织未见表达,3组差异有统计学意义（P〈0.005）。HCC中Bmi1的高表达与年龄、性别、肿瘤大小、肿瘤数目、临床TNM分期、是否有肝硬化及是否有HBsAg感染无显著相关（P〉0.05）,但与组织学分级有关,高、中分化组Bmi1表达率显著高于低分化组（P〈0.05）。肝癌Bmi1阳性组增殖指数（PI）（50.3±21.4）%显著高于阴性表达组（17.3±7.1）%（P〈0.05）,凋亡指数（AI）无明显差异（P〉0.05）。结论：Bmi1在HCC中高表达,其表达增高可能与HCC的进展相关。     

凋亡抑制蛋白XIAP在舌鳞癌组织中表达的研究

目的：研究XIAP在舌鳞癌组织中的表达情况及其与舌鳞癌临床病理特征的关系。方法：采用免疫组织化学SP法检测舌鳞癌组织（36例）以及癌旁正常舌粘膜组织（20例）中XIAP蛋白的表达,并分析其与舌鳞癌临床病理特征的关系。结果：XIAP在舌鳞癌组织中表达阳性率为86．1％,在癌旁正常粘膜组织中阳性率为35％,肿瘤组织中XIAP蛋白阳性率明显高于正常粘膜;不同病理分级及临床分期病例XIAP阳性表达差异无显著性。结论：XIAP蛋白的高表达与舌鳞癌的发生有相关性,可能在舌鳞癌的发生、发展过程中有重要作用。     

乳粘素与半胱氨酸蛋白酶3抑制剂检测舌鳞癌细胞CAL-27凋亡中差异性的比较研究

目的：比较荧光标记的乳粘素（Lactadherin）和半胱氨酸天冬肽酶3（caspase-3）在检测由三氧化二砷（As2O3）诱导舌鳞癌细胞CAL-27凋亡中的敏感性,以期对两种方法在检测结果方面的不同意义有进-步的了解。方法：将配置好终浓度的As20。分别作用舌鳞癌细胞CAL．270h、3h、6h、12h,用荧光标记的caspase-3抑制剂（FAM200—1）标记原位激活的caspase-3,荧光标记的Lactadherin647检测细胞凋亡,荧光倒置显微镜分析其形态学变化,流式细胞仪定量检测并分析其活性变化。结果：As20,诱导细胞3h后．98．5％细胞被FAM200．1标记,只有1．3％细胞被Lactadherin647标记。6h、12h后,被Lactadherin647标记的细胞分别增至2．5％、6．1％。结论：As203作用细胞3h后细胞发生凋亡。FAM200—1先于乳粘素检测出早期凋亡的细胞,caspase-3的激活早于PS翻转。     

Downregulation of miR-503 Promotes ESCC Cell Proliferation,Migration, and Invasion by Targeting Cyclin D1

Esophageal squamous cell carcinoma (ESCC) is one of the most aggressive cancers in China, but the underlying molecular mechanism of ESCC is still unclear. Involvement of microRNAs has been demonstrated in cancer initiation and progression. Despite the reported function of miR-503 in several human cancers, its detailed anti-oncogenic role and clinical significance in ESCC remain undefined. In this study, we examined miR-503 expression by qPCR and found the downregulation of miR-503 expression in ESCC tissue relative to adjacent normal tissues. Further investigation in the effect of miR-503 on ESCC cell proliferation, migration, and invasion showed that enhanced expression of miR-503 inhibited ESCC aggressive phenotype and overexpression of CCND1 reversed the effect of miR-503-mediated ESCC cell aggressive phenotype. Our study further identified CCND1 as the target gene of miR-503. Thus, miR-503 functions as a tumor suppressor and has an important role in ESCC by targeting CCND1.     

沉默DEPDC1抑制鼻咽癌细胞HNE-1和CNE-1侵袭迁移

DEPDC1(DEP domain containing 1)是一个新的肿瘤相关基因,在多种恶性肿瘤的发生发展进程中起着重要作用。我们前期工作中在鼻咽癌细胞内沉默了DEPDC1的表达,发现抑制细胞增殖并诱发细胞凋亡。本研究旨在探讨沉默DEPDC1表达后,对鼻咽癌细胞HNE-1和CNE-1侵袭迁移能力的影响及其分子机制。结果显示,siRNA介导DEPDC1表达沉默后,细胞侧向运动能力、侵袭及迁移能力显著降低。qRT-PCR及Western印迹检测发现DEPDC1沉默导致EMT上游关键转录因子Twist1及间质细胞标志分子Vimentin表达显著下调。这些研究表明,鼻咽癌细胞中DEPDC1通过调节Twist1等EMT关键分子的表达在细胞侵袭转移过程中起关键作用。推测DEPDC1在鼻咽癌中高表达可能对于促进其侵袭转移具有重要作用,进而促进肿瘤发生发展,但具体分子机制仍有待更深入研究。     

宫颈鳞状细胞癌组织LncRNA DCST1-AS1、LncRNA MCM3AP-AS1表达与增殖侵袭基因和预后的关系

摘要 目的：探讨宫颈鳞状细胞癌（CSCC）组织长链非编码核糖核酸（LncRNA）DC-STAMP结构域1-反义1（DCST1-AS1）、LncRNA MCM3AP反义RNA 1（MCM3AP-AS1）表达与增殖侵袭基因和预后的关系。方法：选择2018年1月至2020年1月苏州大学第二附属医院收治的124例CSCC患者,取CSCC组织及其癌旁正常组织,实时荧光定量聚合酶链反应（qRT-PCR）检测LncRNA DCST1-AS1、LncRNA MCM3AP-AS1以及增殖基因（YAP1、Piwil2、EZH2、PKCε）和侵袭基因（Rab11、TUG1、Gli1、FoxM1）mRNA表达。患者出院后随访3年。Pearson相关性分析Lnc RNA DCST1-AS1、Lnc RNA MCM3AP-AS1表达与增殖基因、侵袭基因的相关性。分析Lnc RNA DCST1-AS1、Lnc RNA MCM3AP-AS1表达与临床病理特征的关系。绘制Kaplan-Meier曲线分析预后情况,多因素COX回归分析影响CSCC患者预后的因素。结果：CSCC癌组织中Lnc RNA DCST1-AS1表达高于癌旁组织（P＜0.05）,Lnc RNA MCM3AP-AS1表达低于癌旁组织（P＜0.05）。YAP1、Piwil2、EZH2、PKCε、Rab11、TUG1、Gli1、FoxM1 mRNA水平与CSCC癌组织中Lnc RNA DCST1-AS1表达呈正相关（P＜0.05）,与Lnc RNA MCM3AP-AS1呈负相关（P＜0.05）。低中度分化、FIGO分期Ⅲa期、盆腔淋巴结转移CSCC患者癌组织中Lnc RNA DCST1-AS1表达高于高度分化、FIGO分期Ⅰ～Ⅱ期、无盆腔淋巴结转移宫颈癌患者（P＜0.05）,Lnc RNA MCM3AP-AS1表达低于高度分化、FIGO分期Ⅰ～Ⅱ期、无盆腔淋巴结转移宫颈癌患者（P＜0.05）。Lnc RNA DCST1-AS1高表达、Lnc RNA MCM3AP-AS1低表达CSCC患者3年总生存率低于Lnc RNA DCST1-AS1低表达、Lnc RNA MCM3AP-AS1高表达患者（P＜0.05）。多因素COX回归分析显示盆腔淋巴结转移、Lnc RNA DCST1-AS1高表达是CSCC患者预后不良的危险因素（P＜0.05）,Lnc RNA MCM3AP-AS1高表达是保护因素（P＜0.05）。结论：CSCC组织中LncRNA DCST1-AS1表达上调,LncRNA MCM3AP-AS1表达下调,且与CSCC增殖侵袭和预后不良有关。     

DAPK1、hTERT在口腔鳞癌中的表达及其临床意义研究

目的:探讨凋亡相关蛋白激酶1(DAPK1)、端粒酶催化亚单位(hTERT)在口腔鳞状细胞癌(OSCC)中的表达,并探讨其临床意义。方法:应用免疫组织化学方法检测DAPK1、hTERT在93例OSCC组织及10例癌旁正常组织中的表达,并分析其与OSCC临床病理参数之间的关系及其在OSCC浸润前沿中的作用。结果:癌旁正常组织中DAPK1的表达显著高于OSCC组织中DAPK1的表达,且高、中、低分化OSCC组织中DAPK1的表达比较均有显著差异(P0.05),OSCC浸润前沿组织中DAPK1的表达低于非前沿部分(P0.05),而DAPK1的表达与OSCC浸润前沿的IFG总分无显著相关性(P0.05)。癌旁正常组织中hTERT的表达显著低于OSCC组织中hTERT的表达(P0.05),且高、中、低分化OSCC组织中hTERT的表达比较均有显著差异(P0.05);OSCC浸润前沿组织中hTERT的表达高于非前沿部分(P0.05),且与OSCC浸润前沿的IFG总分有关(P0.05)。DAPK1、hTERT的表达均与OSCC患者的性别、年龄、淋巴结转移无显著相关性(P0.05)。结论:口腔鳞状细胞癌组织,特别是其前沿组织中DAPK1的表达显著下调和hTERT的表达明显上调,可能通过阻碍口腔鳞状细胞癌细胞凋亡,共同促进口腔鳞状细胞癌的生长、分化和浸润。     

Monocarboxylate Transporter 4 Facilitates Cell Proliferation and Migration and Is Associated with Poor Prognosis in Oral Squamous Cell Carcinoma Patients

Monocarboxylate transporter 4 (MCT4) is a cell membrane transporter of lactate. Recent studies have shown that MCT4 is over-expressed in various cancers; however, its role in cancer maintenance and aggressiveness has not been fully demonstrated. This study investigated the role of MCT4 in oral squamous cell carcinoma (OSCC), and found that it is highly expressed in OSCC patients by using immunohistochemistry. Moreover, this over-expression of MCT4 was closely associated with tumor size, TNM classification, lymphatic metastasis, distant metastasis and tumor recurrence, and also poor prognosis. To further study mechanisms of MCT4 in vitro, we used small-interfering RNA to silence its expression in OSCC cell lines. The results showed that knock-down of MCT4 decreased cell proliferation, migration, and invasion. The inhibition of proliferation was associated with down-regulation of p-AKT and p-ERK1/2, while decreased cell migration and invasion may be caused by down-regulation of integrin β4-SRC-FAK and MEK-ERK signaling. Together, these findings provide new insight into the critical role of MCT4 in cell proliferation and metastasis in OSCC.     

Upregulation of the Non-Coding RNA OTUB1-isoform 2 Contributes to Gastric Cancer Cell Proliferation and Invasion and Predicts Poor Gastric Cancer Prognosis

Background: The deubiquitinase OTUB1 plays critical oncogenic roles and facilitates tumor progression in cancer. However, less is known regarding the aberrant expression, clinical significance and biological functions of the non-coding RNA OTUB1-isoform 2. We aimed to evaluate the OTUB1-isoform 2 levels in gastric cancer and their possible correlation with clinicopathologic features and patient survival to reveal its biological effects in gastric cancer progression.Methods: Total RNA extraction was performed on 156 gastric cancer case samples, and RT-qPCR was conducted. Chi-square test analysis was used to calculate the correlation between pathological parameters and the OTUB1-isoform 2 mRNA levels. Kaplan-Meier and Cox proportional hazards analyses were used to analyze the overall survival (OS) and disease-free survival (DFS) rates. Nuclear and cytoplasmic RNAs were isolated to detect the subcellular localization of OTUB1-isoform 2. We also assessed whether overexpression of OTUB1-isoform 2 influenced in vitro cell proliferation, cell cycle progression, tumor cell invasion and migration, as well as in vivo nude mouse xenograft and metastasis models.Results: The OTUB1-isoform 2 expression levels were higher in the gastric cancer samples than in the paratumorous gland samples. OTUB1-isoform 2 expression levels tightly correlated with tumor size, lymph node metastasis and TNM staging. Higher OTUB1-isoform 2 expression levels led to significantly poorer OS and DFS rates, and a multivariate analysis revealed that OTUB1-isoform 2 was an independent risk factor for DFS. OTUB1-isoform 2 was predominantly localized in the cell nucleus. Ectopic overexpression of OTUB1-isoform 2 in gastric cancer cells stimulated proliferation by inducing G₁-S transition, suppression of cell apoptosis and promotion of tumor cell invasion and migration. Finally, OTUB1-isoform 2 overexpression promoted tumor growth and tumor metastasis in nude mice models.Conclusions: Our study suggests that OTUB1-isoform 2 independently predicts poor prognosis and promotes tumor progression in gastric cancer. The non-coding RNA OTUB1-isoform 2 should be targeted in future molecular therapies.     

High Level of COP1 Expression is Associated with Poor Prognosis in Primary Gastric Cancer

COP1 (constitutive photomorphogenic 1, also known as RFWD2) is a p53-targeting E3 ubiquitin ligase containing RING-finger, coiled-coil, and WD40-repeat domains. Recent studies have identified that COP1 is overexpressed in several cancer types and that increased COP1 expression promotes cell proliferation, cell transformation, and tumor progression. In the present study, we investigated the expression and prognostic value of COP1 in primary gastric cancer. To investigate the role of the COP1 gene in primary gastric cancer pathogenesis, real-time quantitative PCR and western blotting were performed to examine COP1 expression in paired cancerous and matched adjacent noncancerous gastric tissues. The results revealed high COP1 mRNA (P=0.030) and protein (P=0.008) expression in most tumor-bearing tissues compared with the matched adjacent non-tumor tissues. The correlated protein expression analysis revealed a negative correlation between COP1 and p53 in gastric cancer samples (P=0.005, r=-0.572). Immunohistochemical staining of gastric cancer tissues from the same patient showed a high COP1 expression and a low p53 expression. To further investigate the clinicopathological and prognostic roles of COP1 expression, we performed immunohistochemical analysis of 401 paraffin-embedded gastric cancer tissue blocks. The data revealed that high COP1 expression was significantly correlated with T stage (P=0.030), M stage (P=0.048) and TNM stage (P=0.022). Consistent with these results, we found that high expression of COP1 was significantly correlated with poor survival in gastric cancer patients (P<0.001). Cox regression analyses showed that COP1 expression was an independent predictor of overall survival (P<0.001). Our data suggest that COP1 could play an important role in gastric cancer and might serve as a valuable prognostic marker and potential target for gene therapy in the treatment of gastric cancer.     

血清应答因子在食管鳞癌细胞侵袭转移中的作用

目的:研究血清应答因子(serum response factor,SRF)在人食管鳞癌细胞体外侵袭转移中的意义。方法:选用EC9706-H、EC9706-L和EC109-H、EC109-L两对高低转移细胞系,采用细胞划痕实验验证食管鳞癌高低转移细胞系体外侵袭转移能力的差异;Western blot检测SRF在两对食管鳞癌高低转移细胞系中的差异表达;在EC9706-H、EC109-H细胞中加入CCG(SRF抑制剂)抑制SRF的表达后,检测其侵袭转移能力的变化。结果:细胞划痕实验验证了两对食管鳞癌高低转移细胞系侵袭转移能力的差异;Western blot结果提示SRF在EC9706-H、EC109-H细胞中的表达水平显著高于EC9706-L、EC109-L细胞;在EC9706-H、EC109-H细胞中加入CCG抑制SRF的表达后,其侵袭转移能力明显减退。结论:SRF在高转移性食管鳞癌细胞系中呈现高表达,在低转移性食管鳞癌细胞系中呈现低表达,抑制高转移性食管鳞癌细胞系中SRF的表达后,其侵袭转移能力下降,提示SRF和食管鳞癌的侵袭转移能力呈正相关。     

miR-101 Inhibiting Cell Proliferation,Migration and Invasion in Hepatocellular Carcinoma through Downregulating Girdin

miR-101 is considered to play an important role in hepato-cellular carcinoma (HCC), but the underlying molecular mechanism remains to be elucidated. Here, we aimed to confirm whether Girdin is a target gene of miR-101 and determine the tumor suppressor of miR-101 through Girdin pathway. In our previous studies, we firstly found Girdin protein was overexpressed in HCC tissues, and it closely correlated to tumor size, T stage, TNM stage and Edmondson-Steiner stage of HCC patients. After specific small interfering RNA of Girdin was transfected into HepG2 and Huh7.5.1 cells, the proliferation and invasion ability of tumor cells were significantly inhibited. In this study, we further explored the detailed molecular mechanism of Girdin in HCC. Interestingly, we found that miR-101 significantly low-expressed in HCC tissues compared with that in matched normal tissues while Girdin had a relative higher expression, and miR-101 was inversely correlated with Girdin expression. In addition, after miR-101 transfection, the proliferation, migration and invasion abilities of HepG2 cells were weakened. Furthermore, we confirmed that Girdin is a direct target gene of miR-101. Finally we confirmed Talen-mediated Girdin knockout markedly suppressed cell proliferation, migration and invasion in HCC while down-regulation of miR-101 significantly restored the inhibitory effect. Our findings suggested that miR-101/Girdin axis could be a potential application of HCC treatment.     

H2-Calponin 在食管鳞癌中的表达及其与食管磷癌预后的关系

目的:探讨H_2-Calponin在食管磷癌组织中的表达及其与食管磷癌患者临床病理特征及预后的相关性。方法:利用免疫组织化学染色技术检测73例食管磷癌及相应癌旁组织中H_2-Calponin的表达情况,并分析其与食管磷癌患者临床病理参数及预后的关系。结果:H_2-calponin在食管磷癌组织中的阳性表达率为95.8%(70/73),显著高于癌旁组织(35.8%,24/67),差异具有统计学意义(p0.0001)。H_2-calponin的表达与食管癌患者的性别、年龄、肿瘤大小、浸润深度及淋巴结转移均无显著相关性,但与AJCC分期显著相关(P0.05)。H_2-calponin的表达水平影响食管癌患者预后及生存期。结论:H_2-Calponin在食管磷癌组织中呈高表达并可预示食管磷癌预后不良。     

LMP1羧基端活化区3对鼻咽癌干细胞SP18迁移与侵袭的影响

为了观察潜伏性膜蛋白1(LMP1)羧基端活性区3(CTAR3)对鼻咽癌干细胞SP18迁移与侵袭的影响,本研究通过建立稳定表达LMP1及CTAR3突变型LMP1(LMP1△252-351)的SP18细胞系(即SP18-LMP1和SP18-LMP1△252-351),观察LMP1-CTAR3缺失突变后对SP18细胞增殖、迁移与侵袭的影响.采用基因芯片分析SP18-LMP1和SP18-LMP1△252-351间的差异表达基因,并验证基因的表达,用生物信息学分析差异表达基因间的相互关系.结果显示:a.SP-LMP1△252-351细胞生长速度较SP-LMP1细胞明显变缓,克隆形成和迁移与侵袭能力降低(n=3,P0.05);b.鉴定出LMP1羧基端CTAR3影响SP18细胞迁移与侵袭的18个基因(其中表达上调基因13个,下调基因5个),经荧光定量PCR验证与基因芯片检测结果基本一致.c.13个差异基因间相互联系,网络节点联系最多的基因是FN1、MMP14、THBS1、ITGA2、IL1B和IL6基因.结果提示,LMP1羧基端CTAR3可能通过调节FN1、MMP14、THBS1、ITGA2、IL1B和IL6基因的表达,发挥其促鼻咽癌干细胞SP18细胞迁移与侵袭的功能.     

SOD2 is a C-myc target gene that promotes the migration and invasion of tongue squamous cell carcinoma involving cancer stem-like cells

Our previous studies revealed that manganese superoxide dismutase (SOD2) contributes to the migration and invasion of tongue squamous cell carcinoma (TSCC). The purpose of the current study was to further clarify the mechanisms of SOD2 in the migration and invasion of TSCC. Side population (SP) cells were used as cancer stem-like cells and further assessed by sphere and colony formation assays, and the expression of stem cell markers (Bmi1, Nanog and ABCG2). We found that UM1 cells (TSCC cells with increased SOD2 expression, migration and invasion abilities) possessed a higher proportion of SP cells, sphere and colony formation, and expressed a higher level of stem cell markers compared to UM2 cells (reduced SOD2 expression, migration and invasion abilities). SOD2 expression as well as migration and invasion abilities were enhanced in SP cells compared to non-SP cells. Knockdown of SOD2 in UM1 cells or SP cells inhibited the migration and invasion abilities, reduced sphere and colony formation, and the expression of stem cell markers. Direct binding of the C-myc protein to the SOD2 promoter was demonstrated by chromatin immunoprecipitation and luciferase assays. Knockdown of C-myc in UM1 cells inhibited SOD2 expression as well as migration and invasion abilities. Our results indicate that cancer stem-like cells play an important role in the migration and invasion of TSCC. SOD2 is a direct target gene of C-myc and C-myc-SOD2-mediated migration and invasion of TSCC involve cancer stem-like cells.     

长链非编码RNA BANCR在食管鳞癌中的表达及对细胞增殖和侵袭能力的影响

目的:探索长链非编码RNA BANCR与食管鳞癌(esophageal squamous cell carcinoma ESCC)临床病理特征以及预后的关系,以及对于ESCC细胞增殖,迁移和侵袭能力的影响。方法:使用实时荧光定量PCR(q RT-PCR)技术检测ESCC组织及多个细胞系中BANCR的表达水平,分析其与临床病理特征及预后的关联,用小干扰RNA(si RNA)干扰BANCR后用CCK8法检测其对ESCC细胞生长的影响,使用transwell法检测对细胞侵袭和转移能力的影响。结果:相对于癌旁组织,有86%(123/142)的癌组织中BANCR表达量升高,BANCR在癌组织中的相对表达水平与肿瘤的组织学分级、TNM分期和淋巴结转移数量相关(P均0.05)。BANCR在本文涉及的八株ESCC细胞中的表达量均高于正常食管上皮细胞(Het1A)。在TE10和KYSE30细胞中敲降BANCR后可明显降低细胞生长速率,并抑制细胞的侵袭和迁移能力(P0.01)。结论:BANCR在ESCC组织和细胞中表达显著上调。并能增强ESCC细胞的增殖和侵袭能力,有希望成为一种新的辅助ESCC早期诊断和预后判断的肿瘤分子标志物。     

Enhanced Expression of ANO1 in Head and Neck Squamous Cell Carcinoma Causes Cell Migration and Correlates with Poor Prognosis

Head and neck squamous cell carcinoma (HNSCC) has the potential for early metastasis and is associated with poor survival. Ano1 (Dog1) is an established and sensitive marker for the diagnosis of gastrointestinal stromal tumors (GIST) and has recently been identified as a Ca(2+) activated Cl(-) channel. Although the ANO1 gene is located on the 11q13 locus, a region which is known to be amplified in different types of human carcinomas, a detailed analysis of Ano1 amplification and expression in HNSCC has not been performed. It is thus still unclear how Ano1 contributes to malignancy in HNSCC. We analyzed genomic amplification of the 11q13 locus and Ano1 together with Ano1-protein expression in a large collection of HNSCC samples. We detected a highly significant correlation between amplification and expression of Ano1 and showed that HNSCC patients with Ano1 protein expression have a poor overall survival. We further analyzed the expression of the Ano1 protein in more than 4'000 human samples from 80 different tumor types and 76 normal tissue types and detected that besides HNSCC and GISTs, Ano1 was rarely expressed in other tumor samples or healthy human tissues. In HNSCC cell lines, expression of Ano1 caused Ca(2+) activated Cl(-) currents, which induced cell motility and cell migration in wound healing and in real time migration assays, respectively. In contrast, knockdown of Ano1 did not affect intracellular Ca(2+) signaling and surprisingly did not reduce cell proliferation in BHY cells. Further, expression and activity of Ano1 strongly correlated with the ability of HNSCC cells to regulate their volume. Thus, poor survival in HNSCC patients is correlated with the presence of Ano1. Our results further suggest that Ano1 facilitates regulation of the cell volume and causes cell migration, which both can contribute to metastatic progression in HNSCC.     

苯胺嘧啶衍生物X-9通过下调整合素β1表达抑制肝细胞癌迁移侵袭

整合素在许多肿瘤细胞中高表达,并且参与肿瘤细胞的侵袭转移。在肝细胞癌中,整合素β1被报导高表达,并促进肿瘤细胞的侵袭。目前,对于整合素的表达调控癌细胞机制以及干预其表达进而抑制肿瘤细胞转移的研究较少。本研究探讨利用小分子化合物抑制整合素表达来抑制肿瘤细胞迁移和侵袭的可能。首先,对临床肝癌细胞患者癌组织和癌旁组织中的整合素β1的表达进行检测,发现其在癌组织中的表达显著高于癌旁组织（P<0.05）。对TCGA肿瘤数据库的生物信息学分析结果同样显示,整合素β1的高表达与肝癌的分期（P=0.019）和预后（P=0.013）相关。通过筛选发现,苯胺嘧啶衍生物X09可以抑制肝癌细胞中整合素β1的mRNA和蛋白质的表达（P<0.01）。细胞划痕愈合实验和细胞穿孔实验结果显示,苯胺嘧啶衍生物X-9能够抑制肝癌细胞的迁移和侵袭（P<0.01）。进一步的研究证实,在肝癌细胞中外源表达整合素β1可以逆转X-9对肝癌细胞迁移和侵袭的抑制;而在敲低整合素β1的细胞中,X-9对细胞的迁移和侵袭的抑制被消除。因此,鉴定出苯胺嘧啶衍生物X-9可以通过下调整合素β1表达,进而抑制肝癌细胞的迁移和侵袭。