LPS对骨肉瘤细胞迁移、侵袭及TLR4/HOTAIR通路的影响

目的探讨脂多糖(LPS)对骨肉瘤细胞迁移和侵袭的影响及其潜在的作用机制。方法将人MG-63骨肉瘤细胞随机分为2组:对照组和LPS组。LPS组细胞用10 g/ml的LPS干预24 h,对照组用生理盐水干预。ELISA检测干预后培养基中促炎因子的水平,Transwell实验检测细胞迁移和侵袭能力,Western Blot检测相关蛋白的表达。结果与对照组相比,LPS组培养基中促炎因子TNF-α、IL-1和IL-6的释放水平均显著增高(P<0.05),LPS组迁移细胞数和侵袭细胞数均显著增高(P<0.05),LPS组中E-cadherin的表达显著降低(P<0.05),而N-cadherin、α-SMA、波形蛋白、TLR4和HOTAIR的表达均显著增高(P<0.05)。结论LPS诱导的肿瘤微环境可促进骨肉瘤细胞的迁移和侵袭,其机制与TLR4/HOTAIR途径介导的EMT过程的发生有密切关系。     

Effect of Diacerein on HOTAIR/IL-6/STAT3, Wnt/β-Catenin and TLR-4/NF-κB/TNF-α axes in colon carcinogenesis

Colorectal cancer (CRC) is a common malignancy with high mortality and poor prognosis. Diacerein (DIA) is an anti-inflammatory used for treatment of osteoarthritis. We delineated some underlying molecular mechanisms of DIA’s anti-carcinogenic effect in CRC using in vivo and in vitro models. Human Caco-2 cells were treated with DIA followed by MTT and Annexin V assays and CRC was experimentally induced using 1,2-dimethylhydrazine. DIA (50 mg/kg/day, orally) was administrated for 8 weeks. The MTT assay confirmed cytotoxic effect of DIA in vitro and Annexin V confirmed its apoptotic effect. DIA resulted in regression of tumour lesions with reduced colonic TLR4, NF-κB and TNF-α protein levels and down-regulated VEGF expression, confirming anti-angiogenic impact. DIA triggered caspase-3 expression and regulated Wnt/β-Catenin pathway, by apparently interrupting the IL-6/STAT3/ lncRNA HOTAIR axis. In conclusion, DIA disrupted IL-6/STAT3/ lncRNA HOTAIR axis which could offer an effective therapeutic strategy for the management of CRC.     

Knockdown of long non-coding RNA HOTAIR inhibits malignant biological behaviors of human glioma cells via modulation of miR-326

Glioma is the most common and aggressive primary adult brain tumor. Long non-coding RNAs (lncRNAs) have important roles in a variety of biological properties of cancers. Here, we elucidated the function and the possible molecular mechanisms of lncRNA HOTAIR in human glioma U87 and U251 cell lines. Quantitative RT-PCR demonstrated that HOTAIR expression was up-regulated in glioma tissues and cell lines. Knockdown of HOTAIR exerted tumor-suppressive function in glioma cells. Further, HOTAIR was confirmed to be the target of miR-326 and miR-326 mediated the tumor-suppressive effects of HOTAIR knockdown on glioma cell lines. Moreover, over-expressed miR-326 reduced the FGF1 expression which played an oncogenic role in glioma by activating PI3K/AKT and MEK 1/2 pathways. In addition, the in vivo studies also supported the above findings. Taken together, knockdown of HOTAIR up-regulated miR-326 expression, and further inducing the decreased expression of FGF1, these results provided a comprehensive analysis of HOTAIR-miR-326-FGF1 axis in human glioma and provided a new potential therapeutic strategy for glioma treatment.     

类风湿关节炎患者血清长链非编码RNA HOTAIR、XIST和H19表达研究

目的探讨3种长链非编码RNA(lncRNA) HOTAIR、XIST和H19在类风湿关节炎(RA)、系统性红斑狼疮(SLE)和健康人对照组血清中的差异表达。方法收取50例RA疾病组、50例SLE疾病对照组和60例健康人对照组血清,共160例血清标本,提取血清总RNA,利用RT-qPCR方法分析HOTAIR、XIST和H19 3种lncRNA表达情况。用Spearman相关性分析探讨RA患者疾病活动指数(DAS28)与HOTAIR表达量的相关性。结果 RA组与健康人对照组和SLE疾病对照组相比,血清HOTAIR表达量显著增加(P0.01),XIST在RA和对照组之间表达量差异无统计学意义; RA患者血清中HOTAIR表达值与DAS28评分呈正相关(R~2=0.27,P=0.002),且DAS28评分≥5.1的疾病高度活动组的HOTAIR表达值显著高于3.2DAS28评分5.1的疾病活动组(P0.01);血清HOTAIR表达量与血清C反应蛋白(CRP)存在正相关关系(R~2=0.192,P=0.002)。结论LncRNA HOTAIR在RA患者血清中的表达量显著性升高且与DAS28评分呈显著正相关关系,血清中HOTAIR有可能成为诊断RA的潜在生物标志物。     

Combined identification of long non-coding RNA CCAT1 and HOTAIR in serum as an effective screening for colorectal carcinoma

Long non-coding RNAs (lncRNAs) CCAT1 and HOTAIR have been shown to play an important regulatory role in cancer biology, and CCAT1 and HOTAIR are upregulated in several cancers, however, its value in the diagnosis of colorectal cancer (CRC) is unclear. Therefore, the aim of this study is to evaluate the clinical significance of plasma CCAT1 and HOTAIR as a biomarker in the screening of CRC. In our study, we found that the levels of HOTAIR (P < 0.05) and CCAT1 (P < 0.05) were significantly higher in plasma of CRC patients than that of the healthy control. Moreover, the levels of lincRNA-p21 (P < 0.05) were obviously decreased in plasma of CRC patients as compared to those of healthy control. There was highly correlated for CCAT1 (R = 0.752, mean differences = -0.06 ± 1.20), HOTAIR (R = 0.739, mean differences = -0.26 ± 0.76) and lincRNA-p21 (R = 0.848, mean differences = -0.41 ± 0.89) in plasma and serum. By receiver operating characteristic curve (ROC) analysis, plasma CCAT1 provided the higher diagnostic performance for detection of CRC (the area under the ROC curve (AUC), 0.836; P < 0.001; sensitivity, 75.7%; specificity, 85.3%). Moreover, CCAT1 combining with HOTAIR could provide a more effective diagnosis performance (AUC, 0.954, P < 0.001, sensitivity, 84.3%; specificity, 80.2%). Most importantly, this combination was effective to detect CRC at an early stage (85%). In conclusion, our results demonstrated that increased plasma HOTAIR and CCAT1 could be used as a predictive biomarker for CRC screening, and that combination of HOTAIR and CCAT1 had a higher positive diagnostic rate of CRC than HOTAIR or CCAT1 alone.     

长链非编码RNA HOTAIR在膀胱癌中的表达及临床意义

目的: 探讨长链非编码RNA HOTAIR在膀胱癌组织与癌旁正常组织的表达差异,了解HOTAIR与膀胱癌恶性程度、进展及预后的相关性。方法: 采用qRT-PCR方法检测88例膀胱癌组织与癌旁正常组织中HOTAIR RNA的差异表达,分析HOTAIR表达水平与患者临床病理特征的关系,采用Kapaln-Meier生存分析及COX 比例风险回归模型分析HOTAIR表达水平与膀胱患者预后的相关性。结果: 膀胱癌组织中HOTAIR RNA 表达明显高于癌旁正常组织,HOTAIR RNA高表达与肿瘤高分级及临床高分期有关（P<0.05）,HOTAIR高表达组膀胱癌患者5年生存率明显低于HOTAIR低表达组（P<0.05）,COX 回归分析提示HOTAIR RNA高表达可能为膀胱癌不良预后的独立危险因子之一。结论: 长链非编码RNA HOTAIR与膀胱癌发生发展及其恶性程度相关,HOTAIR可以作为预测膀胱癌患者生存率的分子标志物。     

Clinical significance of HOTAIR expression in colon cancer

AIM: To detect the expression of the long noncoding RNA HOTAIR in colon cancer and analyze its relationship with clinicopathological parameters of colon cancer. METHODS: Total RNA was extracted from 80 colon cancer tissues and matched tumor-adjacent normal colon tissues and reverse transcribed. Quantitative polymerase chain reaction was used to detect the expression of HOTAIR. The relationship between the expression of HOTAIR and clinicopathological parameters of colon cancer was analyzed. RESULTS: The expression of HOTAIR was significantly higher in colon cancer tissues than in matched tumoradjacent normal colon tissues(P 0.05). HOTAIR expression was significantly higher in cases with lymph node metastasis than in those without metastasis; in lowly differentiated and undifferentiated cases than in highly and moderately differentiated cases; and in stages Ⅲ + Ⅳ cases than in stages?Ⅰ?+ Ⅱ cases(P 0.05).CONCLUSION: HOTAIR expression is upregulated in colon cancer, suggesting that HOTAIR plays an important role in the tumorigenesis, development and metastasis of colon cancer. HOTAIR may act as an oncogene and represents a new molecular target for the treatment of colon cancer.     

肾癌细胞系中HOTAIR表达的测定及稳定高表达HOTAIR肾癌细胞株的建立

目的:检测常用肾癌细胞系中HOTAIR的表达情况及建立稳定高表达HOTAIR肾癌细胞株?方法:①q-PCR检测正常肾细胞系(HK-2)和4种肾癌细胞系(os-rc-2,kevt-3,achn和769-p)中HOTAIR的表达;②利用携带HOTAIR基因的慢病毒(pLenti GFP Puro-HOTAIR)感染肾癌细胞系(769-p)后,并采用嘌呤霉素(Puro)抗性筛选方法建立高表达HOTAIR肾癌细胞株?结果:①5种细胞系HOTAIR表达的高低依次是kevt-3?os-rc-2?achn?769-p?HK-2;②慢病毒(pLenti GFP Puro-HOTAIR)转染肾癌细胞系(769-p)后,HOTAIR表达明显升高?结论:①高侵袭性肾癌细胞株(kevt-3?os-rc-2)中HOTAIR的表达明显高于低侵袭性肾癌细胞株(achn?769-p)及正常肾细胞株(HK-2);②成功建立稳定表达的HOTAIR的769-p-HOTAIR细胞系?