Summary: This study aimed to examine the effect of long non-coding RNA (LncRNA) MEG3 on the biological behaviors of renal cell carcinoma (RCC) cells 786-0 and the possible mechanism. MEG3 expression levels were d...Summary: This study aimed to examine the effect of long non-coding RNA (LncRNA) MEG3 on the biological behaviors of renal cell carcinoma (RCC) cells 786-0 and the possible mechanism. MEG3 expression levels were detected by RT-qPCR in Rmaor tissues and adjacent non-tumor tissues from 29 RCC patients and in RCC lines 786-0 and SN12 and human embryonic kidney cell line 293T. Plasmids GV144-MEG3 (MEG3 overexpression plasmid) and GV144 (control plasmid) were stably transfected into 786-0 cells by using lipofectamine 2000. Cell viabilities were determined by MTT, cell apoptosis rates by flow cytometry following PE Annexin V and 7AAD staining, apoptosis-related protein expressions by Western blotting, and Bcl-2 mRNA by RT-qPCR in the transfected cells. The results showed that MEG3 was evidently downregulated in RCC tissues (P〈0.05) and RCC cell lines (P〈0.05). The viabilities of 786-0 cells were decreased significantly after transfection with GV144-MEG3 for over 24 h (P〈0.05). Consistently, the apoptosis rate was significantly increased in 786-0 cells transfected with GV144-MEG3 for 48 h (P〈0.05). Furthermore, overexpression of MEG3 could reduce the expression of Bcl-2 and procaspase-9 proteins, enhance the expression of cleaved caspase-9 protein, and promote the release of cytochrome c protein to cytoplasm (P〈0.05). Additionally, Bcl-2 mRNA level was declined by MEG3 overexpression (P〈0.05). It was concluded that MEG3 induces the apoptosis of RCC cells possibly by activating the mitochondrial pathway.展开更多
非编码RNA(non-coding RNA,ncRNA)为一类不编码蛋白的RNA分子,是机体重要的生物调控因子,在转录及转录后水平调控基因表达,影响糖尿病血管病变的发展过程。ncRNA按其片段大小主要分为微RNA(microRNA,miRNA)、长链非编码RNA(long non cod...非编码RNA(non-coding RNA,ncRNA)为一类不编码蛋白的RNA分子,是机体重要的生物调控因子,在转录及转录后水平调控基因表达,影响糖尿病血管病变的发展过程。ncRNA按其片段大小主要分为微RNA(microRNA,miRNA)、长链非编码RNA(long non coding RNA,lncRNA)及环状RNA(circular RNA,circRNA)。miRNA可在转录后水平调控靶基因表达,并有成为临床诊断标志物的潜能。lncRNA影响多种分子信号通路,其在糖尿病血管病变中的作用逐渐受到关注。circRNA具有显著的基因调节功能,可与miRNA竞争结合位点,参与调控糖尿病血管病变。该文回顾目前有关ncRNA与糖尿病血管病变的研究,探讨ncRNA与糖尿病微血管及大血管病变间的关系,为糖尿病血管病变的诊断和治疗提供新思路。展开更多
目的:分析长链非编码RNA(long non-coding RNA,lnc RNA)BC200在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的表达情况,并探讨其临床意义及其与上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白E-cadherin、N-...目的:分析长链非编码RNA(long non-coding RNA,lnc RNA)BC200在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的表达情况,并探讨其临床意义及其与上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白E-cadherin、N-cadherin及Snail之间的关系。方法:采用实时荧光定量PCR(QPCR)法检测2014年1月至2018年6月天津市胸科医院60例癌组织及60例对应的正常肺组织中BC200表达情况,40例癌组织及40例正常肺组织中E-cadherin、N-cadherin及Snail m RNA表达水平,统计分析NSCLC中BC200与E-cadherin、N-cadherin及Snail m RNA表达水平以及两者的相关性,分析BC200与临床病理特征(性别、年龄、TNM分期、肿瘤大小、淋巴结转移及病理类型水平)的关系,应用受试者工作特征(receiver operating characteristic curve,ROC)曲线鉴别诊断NSCLC的效能;采用免疫组织化学法检测E-cadherin、N-cadherin及Snail蛋白在40例NSCLC组织及20例正常肺组织中表达情况,分析其与BC200表达情况的关系。结果:1)肺癌组织中的BC200、N-cadherin及Snail m RNA的表达水平显著高于正常肺组织中的表达水平(P<0.05),E-cadherin m RNA的表达量低于正常肺组织(P<0.05);2)E-cadherin、N-cadherin及Snail在NSCLC组织中阳性表达率分别为40.0%(16/40)、57.5%(23/40)、57.5%(23/40),在正常肺组织中的阳性表达率为95%(19/20)、5%(1/20)、10%(2/20),表达差异具有统计学意义(P<0.05);3)肺癌组织中BC200的高表达与淋巴结转移、临床分期、E-cadherin、N-cadherin及Snail阳性率相关(P<0.05),BC200的表达与E-cadherin m RNA的表达呈负相关(r=-0.31,P<0.05),与Snail、N-cadherin m RNA的表达呈正相关(r=0.305、r=0.257,P<0.05);4)ROC曲线分析表明,组织lnc RNA表达水平对NSCLC的诊断敏感度和特异度均较满意。结论:BC200在NSCLC组织中高表达,且与淋巴结转移、临床分期、E-cadherin、N-cadherin及Snail阳性率相关,BC200的表达与E-cadherin m RNA的表达呈负相关,与Snail、N-cadherin m 展开更多
基金supported by grants from the National Natural Science Foundation of China(Nos.81001132,81172423,and 81272816)
文摘Summary: This study aimed to examine the effect of long non-coding RNA (LncRNA) MEG3 on the biological behaviors of renal cell carcinoma (RCC) cells 786-0 and the possible mechanism. MEG3 expression levels were detected by RT-qPCR in Rmaor tissues and adjacent non-tumor tissues from 29 RCC patients and in RCC lines 786-0 and SN12 and human embryonic kidney cell line 293T. Plasmids GV144-MEG3 (MEG3 overexpression plasmid) and GV144 (control plasmid) were stably transfected into 786-0 cells by using lipofectamine 2000. Cell viabilities were determined by MTT, cell apoptosis rates by flow cytometry following PE Annexin V and 7AAD staining, apoptosis-related protein expressions by Western blotting, and Bcl-2 mRNA by RT-qPCR in the transfected cells. The results showed that MEG3 was evidently downregulated in RCC tissues (P〈0.05) and RCC cell lines (P〈0.05). The viabilities of 786-0 cells were decreased significantly after transfection with GV144-MEG3 for over 24 h (P〈0.05). Consistently, the apoptosis rate was significantly increased in 786-0 cells transfected with GV144-MEG3 for 48 h (P〈0.05). Furthermore, overexpression of MEG3 could reduce the expression of Bcl-2 and procaspase-9 proteins, enhance the expression of cleaved caspase-9 protein, and promote the release of cytochrome c protein to cytoplasm (P〈0.05). Additionally, Bcl-2 mRNA level was declined by MEG3 overexpression (P〈0.05). It was concluded that MEG3 induces the apoptosis of RCC cells possibly by activating the mitochondrial pathway.
文摘目的:分析长链非编码RNA(long non-coding RNA,lnc RNA)BC200在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的表达情况,并探讨其临床意义及其与上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白E-cadherin、N-cadherin及Snail之间的关系。方法:采用实时荧光定量PCR(QPCR)法检测2014年1月至2018年6月天津市胸科医院60例癌组织及60例对应的正常肺组织中BC200表达情况,40例癌组织及40例正常肺组织中E-cadherin、N-cadherin及Snail m RNA表达水平,统计分析NSCLC中BC200与E-cadherin、N-cadherin及Snail m RNA表达水平以及两者的相关性,分析BC200与临床病理特征(性别、年龄、TNM分期、肿瘤大小、淋巴结转移及病理类型水平)的关系,应用受试者工作特征(receiver operating characteristic curve,ROC)曲线鉴别诊断NSCLC的效能;采用免疫组织化学法检测E-cadherin、N-cadherin及Snail蛋白在40例NSCLC组织及20例正常肺组织中表达情况,分析其与BC200表达情况的关系。结果:1)肺癌组织中的BC200、N-cadherin及Snail m RNA的表达水平显著高于正常肺组织中的表达水平(P<0.05),E-cadherin m RNA的表达量低于正常肺组织(P<0.05);2)E-cadherin、N-cadherin及Snail在NSCLC组织中阳性表达率分别为40.0%(16/40)、57.5%(23/40)、57.5%(23/40),在正常肺组织中的阳性表达率为95%(19/20)、5%(1/20)、10%(2/20),表达差异具有统计学意义(P<0.05);3)肺癌组织中BC200的高表达与淋巴结转移、临床分期、E-cadherin、N-cadherin及Snail阳性率相关(P<0.05),BC200的表达与E-cadherin m RNA的表达呈负相关(r=-0.31,P<0.05),与Snail、N-cadherin m RNA的表达呈正相关(r=0.305、r=0.257,P<0.05);4)ROC曲线分析表明,组织lnc RNA表达水平对NSCLC的诊断敏感度和特异度均较满意。结论:BC200在NSCLC组织中高表达,且与淋巴结转移、临床分期、E-cadherin、N-cadherin及Snail阳性率相关,BC200的表达与E-cadherin m RNA的表达呈负相关,与Snail、N-cadherin m