奥沙利铂通过调控miRNA-7-5p/RAF-1促进胃癌SGC-7901细胞凋亡的研究

目的探讨奥沙利铂如何调控MAPK通路,抑制胃癌细胞的增殖。方法 NCBI检索文献,利用TargetScan、StarBase和miRBase数据库,进行GO分析与KEGG通路富集,找到相关miRNAs,预测靶基因。应用Real-time PCR、MTT、Hoechst33258、流式细胞术、细胞划痕实验、Western blot等方法分析人胃癌SGC-7901细胞的增殖、细胞周期、侵袭及蛋白表达情况。结果胃癌细胞中miR-7-5p显著低表达,RAF1与miR-7-5p存在互靶关系。miR-7-5p mimics与奥沙利铂均可促进SGC-7901细胞的凋亡,提高G1期细胞百分率(P<0.05),降低侵袭、迁移速度。caspase3、caspase9蛋白表达升高,Bcl-2/Bax比值降低(P<0.05)。结论过表达mi R-7-5p与奥沙利铂均可促进胃癌SGC-7901细胞的凋亡,提示奥沙利铂可能通过上调mi RNA-7-5p促进SGC-7901细胞的凋亡,降低侵袭、迁移速度。     

靶向沉默髓鞘转录因子MyT1对脑胶质瘤细胞恶性生物学行为的影响

目的探讨靶向沉默髓鞘转录因子1(MyT1)对人脑胶质瘤细胞迁移、侵袭和黏附的影响和相关分子机制。方法设计特异性靶向沉默MyT1基因的shRNA,包装慢病毒后感染人脑胶质瘤U-118MG和U-87MG细胞,qPCR和Western blot检测两种细胞中MyT1的表达水平,BrdU实验、细胞划痕实验、Transwell实验和细胞黏附实验分别检测两种细胞的迁移、侵袭和黏附能力的变化,qPCR检测细胞黏附和肿瘤转移相关基因的表达水平。结果在HEK293T细胞中包装了特异性靶向MyT1基因的shRNA慢病毒,并成功感染了U-118MG和U-87MG细胞;两种细胞中MyT1 mRNA和蛋白表达水平均显著下调(均P<0.05),细胞的迁移、侵袭和黏附能力均有一定程度的降低(均P<0.05),细胞黏附相关基因表达水平显著下降,肿瘤转移相关基因表达水平显著上升(均P<0.05)。结论靶向沉默MyT1基因对人脑胶质瘤U-118MG和U-87MG细胞的迁移、侵袭和黏附能力有抑制作用,其机制可能与调控细胞黏附和肿瘤转移相关基因的表达有关。MyT1基因的表达,可能是脑胶质瘤诊疗的一个潜在靶标...     

波形蛋白对前列腺癌细胞侵袭与转移的影响

背景与目的:波形蛋白(vimentin)是一种细胞骨架蛋白,参与调节细胞的运动和增殖。本研究通过检测波形蛋白在前列腺癌细胞系中的表达,探讨其对前列腺癌细胞侵袭与转移的影响。方法:应用二维电泳-质谱分析(two-dimensionalgel electrophoresis matrix-assisted laser desorption/time of flight mass spectrometry,2-DE MALDI TOF-MS)检测波形蛋白在前列腺癌配对细胞系中的差异表达,用基因干预技术结合体外侵袭实验探讨波形蛋白对细胞侵袭能力的影响。结果:波形蛋白在前列腺癌高转移细胞系PC-3M-1E8中的表达高于低转移细胞系PC-3M-2B4中的表达。成功构建波形蛋白反义真核表达载体和正义真核表达载体,分别转染PC-3M-1E8和PC-3M-2B4细胞,转染波形蛋白反义真核表达载体的细胞(PC-3M-1E8/vas)的穿膜细胞数为99.3±4.8,明显低于空质粒对照组细胞[PC-3M-1E8/3.1(-)]的319.4±6.5(P<0.01);转染波形蛋白正义真核表达载体的细胞(PC-3M-2B4/vs)的穿膜细胞数为330.5±5.8,明显高于空质粒对照组细胞[PC-3M-2B4/3.1( )]的98.6±7.5(P<0.01)。结论:波形蛋白高表达可促进前列腺癌细胞的侵袭与转移。     

ERCC 1在前列腺癌PC-3细胞和组织样本中的表达及其临床意义

目的 探讨切除修复交叉互补基因1(ERCC 1)在前列腺癌（PCa）PC-3细胞和前列腺样本中的表达情况及与预后的关系。方法 Western blot检测小干扰核糖核酸（siRNA）ERCC 1在转染PC-3细胞后ERCC 1蛋白的表达水平,MTT法检测细胞增殖活力,Transwell试验检测细胞迁移和侵袭能力。免疫组化（IHC）检测80例PCa组织及30例前列腺增生（BPH）组织中ERCC 1蛋白的表达水平,分析ERCC 1与PCa临床病理特征及其预后关系。结果 siRNA ERCC 1质粒转染PC-3细胞后Western blot检测证实ERCC 1表达水平明显减低。Transwell试验结果显示siRNA干扰表达ERCC 1后PC-3细胞迁移和侵袭能力下降,差异有统计学意义（P<0.05）。IHC结果提示ERCC 1在PCa样本中阳性表达率为71.3%(57/80),高表达率为23.8%(19/80,IRS≥6);在BPH样本中阳性表达率为10%(3/30),均为低表达（IRS<6）。ERCC 1表达与PCa患者术前PSA值,Gleason评分,病理分期（pT）,淋巴...     

Survivin和MMP-2在老年宫颈癌的表达及与肿瘤侵袭转移的关系

目的探讨Survivin和基质金属蛋白酶-2(MMP-2)在老年宫颈癌的表达及与肿瘤侵袭转移的关系。方法选择72例老年宫颈上皮内瘤病变(CIN)患者及58例老年宫颈癌患者为研究对象,采用免疫组织化学方法检测患者病变组织Survivin和MMP-2的表达。比较CIN患者与宫颈癌患者Survivin和MMP-2表达阳性率的差别,并探讨Survivin和MMP-2与老年宫颈癌肿瘤侵袭转移的关系。结果 CINⅠ～Ⅱ级、CINⅢ级及宫颈癌患者Survivin和MMP-2阳性率依次升高(P<0.05);Survivin和MMP-2阳性率与FIGO分期、淋巴转移、间质浸润有关(P<0.05)。结论不同级别宫颈病变Survivin和MMP-2阳性率不同,Survivin和MMP-2在老年宫颈癌患者肿瘤侵袭转移等生物学行为中起重要作用。     

侵袭性垂体腺瘤的诊治及其护理

目的 探讨侵袭性垂体腺瘤的临床特点与护理措施.方法 回顾分析62例侵袭性垂体腺瘤的临床表现、影像学表现及治疗效果,并结合同期78例非侵袭性垂体腺瘤的临床资料进行对比研究.结果 大腺瘤和巨大腺瘤的侵袭性腺瘤发生率(63.8%)明显高于微腺瘤和小腺瘤(15.9%),差异有统计学意义(P<0.01);功能性腺瘤和非功能性腺瘤中的侵袭性腺瘤发生率分别为44.8%,43.4%,差异无统计学意义(P>0.05);侵袭性腺瘤中肿瘤卒中的发生率(22.6%)明显高于非侵袭性腺瘤(5.1%);侵袭性腺瘤术后复发率(16.1%)明显高于非侵袭性腺瘤(3.8%);侵袭性腺瘤的手术全切率(46.8%)明显低于非侵袭性腺瘤(88.5%),有统计学意义(P<0.01).结论 侵袭性垂体腺瘤的诊断应结合影像学检查、术中所见和病理学检查来综合判定.手术全切率低,术后复发率高,治疗上应采取手术治疗结合放射治疗和药物治疗的综合治疗.加强对侵袭性垂体腺瘤术后的护理,可有效预防和减轻经蝶侵袭性垂体腺瘤术后并发症.     

基质金属蛋白酶9抑制剂对人舌鳞癌SCC-25细胞侵袭、迁移能力的影响及机制探讨

目的:探讨基质金属蛋白酶9(MMP-9)抑制剂对人舌鳞状细胞癌(TSCC)SCC-25细胞侵袭、迁移能力的影响及其相关机制.方法:取对数生长期SCC-25细胞,分别采用0、5、10、20μmol/L的MMP-9抑制剂BB-94干预,分别设为对照组、实验A组、实验B组和实验C组.采用RT-qPCR和Western免疫印迹...     

RING finger and WD repeat domain 3 regulates proliferation and metastasis through the Wnt/β-catenin signalling pathways in hepatocellular carcinoma

BACKGROUNDHepatocellular carcinoma (HCC) exhibits high invasiveness and mortality rates, and the molecular mechanisms of HCC have gained increasing research interest. The abnormal DNA damage response has long been recognized as one of the important factors for tumor occurrence and development. Recent studies have shown the potential of the protein RING finger and WD repeat domain 3 (RFWD3) that positively regulates p53 stability in response to DNA damage as a therapeutic target in cancers. AIMTo investigate the relationship between HCC and RFWD3 in vitro and in vivo and explored the underlying molecular signalling transduction pathways. METHODS RFWD3 gene expression was analyzed in HCC tissues and adjacent normal tissues. Lentivirus was used to stably knockdown RFWD3 expression in HCC cell lines. After verifying the silencing efficiency, Celigo/cell cycle/apoptosis and MTT assays were used to evaluate cell proliferation and apoptosis. Subsequently, cell migration and invasion were assessed by wound healing and transwell assays. In addition, transduced cells were implanted subcutaneously and injected into the tail vein of nude mice to observe tumor growth and metastasis. Next, we used lentiviral-mediated rescue of RFWD3 shRNA to verify the phenotype. Finally, the microarray, ingenuity pathway analysis, and western blot analysis were used to analyze the regulatory network underlying HCC. RESULTSCompared with adjacent tissues, RFWD3 expression levels were significantly higher in clinical HCC tissues and correlated with tumor size and TNM stage (P < 0.05), which indicated a poor prognosis state. RFWD3 silencing in BEL-7404 and HCC-LM3 cells increased apoptosis, decreased growth, and inhibited the migration in shRNAi cells compared with those in shCtrl cells (P < 0.05). Furthermore, the in vitro results were supported by the findings of the in vivo experiments with the reduction of tumor cell invasion and migration. Moreover, the rescue of RFWD3 shRNAi resulted in the resumption of invasion and metastasis in HCC cell lines. Finally, gene expression profiling and subsequent experimental verification revealed that RFWD3 might influence the proliferation and metastasis of HCC via the Wnt/β-catenin signalling pathway.CONCLUSIONWe provide evidence for the expression and function of RFWD3 in HCC. RFWD3 affects the prognosis, proliferation, invasion, and metastasis of HCC by regulating the Wnt/β-catenin signalling pathway.     

Epidermal growth factor-like domain 7 promotes cell invasion and angiogenesis in pancreatic carcinoma

Epidermal growth factor-like domain 7 (EGFL7), also known as vascular endothelial stain, was firstly identified as a modulator of smooth muscle cell migration. Though the expression of EGFL7 was reported to be up-regulated during tumorigenesis, the clinical and biological functions of EGFL7 in pancreatic carcinoma (PC) were still not fully elucidated. In this study, we found that the serum EGFL7 level in PC tissues was statistically higher than that in normal subjects (p < 0.001), and its level in non-resectable patients was also higher than that in resectable ones (p = 0.013). Among these resectable PC patients, the postoperative EGFL7 expression was significantly down-regulated when tumors were resected (p = 0.018). Using the immunohistochemistry method, our results demonstrated that the positive expression of EGFL7 was significantly associated with the TNM stage (p = 0.024), lymph node metastasis (p = 0.003) and local invasion (p = 0.022), and the EGFL7 expression closely correlated to the micro-vessel density (MVD) in PC tissues by Spearman analysis (r = 0.941, p = 0.000). In vitro, EGFL7 was silenced by the small interference RNA in PC cells, and our data indicated that down-regulation of EGFL7 did not influence the cycle progression, proliferation, colony formation and apoptosis of PC cells (p > 0.05), whereas inhibition of EGFL7 expression could decrease PaCa-2 cell invasion (p < 0.05). More interestingly, by tubular formation, Chick embryo chorioallantoic membrane (CAM) and ELISA assays, our results revealed that silencing EGFL7 expression represented a strong inhibiting effect on tubular formation of micro-vessels through down-regulating the protein levels of VEGF and Ang-2 (p < 0.05). Our results raised the possibility of using EGFL7 as a potential prognostic biomarker and therapy target of PC, and down-regulation of EGFL7 might be considered to be a potentially important molecular treatment strategy for patients with PC.     

Impact of 3′UTR variation patterns of the KRAS gene on the aggressiveness of pancreatobiliary tumors with the KRAS G13D mutation in a Turkish population

ObjectiveSeveral studies have demonstrated the importance of mutations in codons 12, 13 and 61 and variations in the 3′ untranslated region (3′UTR) of the KRAS gene, frequently observed genetic events in the progression of pancreatobiliary tumors (PBT). However, limited data exist on the clinical effect of these alterations. The aim of the current study was to clarify the frequency of relevant alterations of the 3′UTR regions of the KRAS gene and the effect of KRAS 3′UTR polymorphisms on the prognosis of patients with codon 12, 13 and 61 mutations in a Turkish population with PBT.MethodsCodons 12, 13, and 61 and 3′UTRs of the KRAS gene were screened by single-strand conformation polymorphism (SSCP) analysis and DNA sequencing in 43 patients and 10 controls. Chi-squared and independent sample T tests were used to evaluate the results of the mutation analysis and clinical features of the patients.ResultsWe defined the c.38G > A (rs112445441, p.G13D) (39.54%) mutation and two 3′UTR variations, c.*4066delA (rs560890523) (23.26%) and c.*4065_*4066delAA (rs57698689) (6.98%), in the KRAS gene of Turkish patients. There was a statistically significant relationship between the c.*4066delA (rs560890523) and c.*4065_*4066delAA (rs57698689) variations and invasion and lymph node metastasis status of the patients (p < 0.001). Compared to patients with c.38G > A (rs112445441, p.G13D), patients with c.*4066delA (rs560890523) and c.38G > A (rs112445441, p.G13D) presented more aggressive tumors with highly invasive features. The present study contributes findings regarding the clinical effects of KRAS alterations in PBT. Based on our study, further investigations are required.