Characterization of N6-methyladenosine long non-coding RNAs in sporadic congenital cataract and age-related cataract

AIM:To characterize the N6-methyladenosine(m6A)modification patterns in long non-coding RNAs(lncRNAs)in sporadic congenital cataract(CC)and age-related cataract(ARC).METHODS:Anterior capsule of the lens were collected from patients with CC and ARC.Methylated RNA immunoprecipitation with next-generation sequencing and RNA sequencing were performed to identify m6A-tagged lncRNAs and lncRNAs expression.Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses and Gene Ontology annotation were used to predict potential functions of the m6A-lncRNAs.RESULTS:Large amount of m6A peaks within lncRNA were identified for both CC and ARC,while the level was much higher in ARC(49870 peaks)than that in CC(18688 peaks),yet those difference between ARC in younger age group(ARC-1)and ARC in elder age group(ARC-2)was quite slight.A total of 1305 hypermethylated and 1178 hypomethylated lncRNAs,as well as 182 differential expressed lncRNAs were exhibited in ARC compared with CC.On the other hand,5893 hypermethylated and 5213 hypomethylated lncRNAs,as well as 155 significantly altered lncRNA were identified in ARC-2 compared with ARC-1.Altered lncRNAs in ARC were mainly associated with the organization and biogenesis of intracellular organelles,as well as nucleotide excision repair.CONCLUSION:Our results for the first time present an overview of the m6A methylomes of lncRNA in CC and ARC,providing a solid basis and uncovering a new insight to reveal the potential pathogenic mechanism of CC and ARC.

Long non-coding RNA GATA6-AS1 is mediated by N6-methyladenosine methylation and inhibits the proliferation and metastasis of gastric cancer

BACKGROUND Through experimental research on the biological function of GATA6-AS1,it was confirmed that GATA6-AS1 can inhibit the proliferation,invasion,and migration of gastric cancer cells,suggesting that GATA6-AS1 plays a role as an anti-oncogene in the occurrence and development of gastric cancer.Further experi-ments confirmed that the overexpression of fat mass and obesity-associated protein(FTO)inhibited the expression of GATA6-AS1,thereby promoting the occurrence and development of gastric cancer.AIM To investigate the effects of GATA6-AS1 on the proliferation,invasion and migration of gastric cancer cells and its mechanism of action.METHODS We used bioinformatics methods to analyze the Cancer Genome Atlas(https://portal.gdc.cancer.gov/.The Cancer Genome Atlas)and download expression data for GATA6-AS1 in gastric cancer tissue and normal tissue.We also constructed a GATA6-AS1 lentivirus overexpression vector which was transfected into gastric cancer cells to investigate its effects on proliferation,migration and invasion,and thereby clarify the expression of GATA6-AS1 in gastric cancer and its biological role in the genesis and development of gastric cancer.Next,we used a database(http://starbase.sysu.edu.cn/starbase2/)to analysis GATA6-AS1 whether by m6A methylation modify regulation and predict the methyltransferases that may methylate GATA6-AS1.Furthermore,RNA immunoprecipitation experiments confirmed that GATA6-AS1 was able to bind to the m6A methylation modification enzyme.These data allowed us to clarify the ability of m6A methylase to influence the action of GATA6-AS1 and its role in the occurrence and development of gastric cancer.RESULTS Low expression levels of GATA6-AS1 were detected in gastric cancer.We also determined the effects of GATA6-AS1 overexpression on the biological function of gastric cancer cells.GATA6-AS1 had strong binding ability with the m6A demethylase FTO,which was expressed at high levels in gastric cancer and negatively correlated with the expression of GATA6-AS1.Following transfection with siRNA to knock down the expression of FTO,the expression levels of GATA6-AS1 were up-regulated.Finally,the proliferation,migration and invasion of gastric cancer cells were all inhibited following the knockdown of FTO expression.CONCLUSION During the occurrence and development of gastric cancer,the overexpression of FTO may inhibit the expression of GATA6-AS1,thus promoting the proliferation and metastasis of gastric cancer.

长链非编码RNA HOTAIR对非小细胞肺癌迁移与侵袭能力的影响

目的:探讨非小细胞肺癌组织中长链非编码RNA HOTAIR的表达情况及其对细胞侵袭与迁移水平的作用。方法:利用定量的反转录PCR技术检测50例非小细胞肺癌组织中LncRNA HOTAIR的表达,然后通过过表达技术与RNA干扰技术研究LncRNA HOTAIR的主要功能。再对其pcDNA-HOTAIR与si-HOTAIR进行转染调控LncRNA HOTAIR的表达,设置空白载体与阴性对照组,应用反转录PCR进行转染效率的定量检测。然后分别实施MTT与Transwell两种方法来判断LncRNA HOTAIR的异常表达对非小细胞肺癌细胞活性的干预作用。结果:在非小细胞肺癌组织中LncRNA HOTAIR的相对表达水平是(24.50±7.43)。LncRNA HOTAIR在SPC-A1和SK-MES-1两种细胞中的相对表达水平比在正常的支气管上皮细胞中更高,在A549细胞中的相对表达水平比在正常的支气管上皮细胞中更低。而经2d的转染siRNA后,LncRNA HOTAIR在A549与SPC-A1两种细胞中的表达水平降低;pcDNA3.1-HOTAIR后,LncRNA HOTAIR在A549细胞中的表达水平升高。利用RNA干扰技术阻碍HOTAIR的表达水平,不能抑制非小细胞肺癌细胞的增殖。而利用si-HOTAIR的转染降低LncRNA HOTAIR的表达水平能够阻碍癌细胞的迁移与侵袭作用(P<0.05);而LncRNA HOTAIR的过度表达能够明显推动癌细胞的迁移与侵袭作用(P<0.05)。结论:HOTAIR在非小细胞肺癌中的高水平异常表达,能够提高非小细胞肺癌细胞迁移与侵袭水平,可能和患者的不良预后有联系。

Construction of a prognostic signature of m7G-related lncRNAs in bladder cancer:a bioinformatics analysis

Background:N7-methylguanosine(m7G)-related plays an important role in the occurrence and development of tumors,and some recent studies have pointed out that long non-coding RNA is involved in the occurrence and development of various cancers.However,there is no literature on how m7G-related lncRNAs predict the prognosis of bladder cancer.The purpose of this study was to develop a predictive feature based on long non-coding RNA(lncRNAs)associated with m7G to predict the prognosis of patients with bladder cancer.Methods:We obtained the RNA transcriptome data and clinical data of bladder cancer patients through the cancer genome atlas database,and obtained the lncRNAs related to m7G by co-expression analysis and Cox regression analysis.Then the signature was evaluated by receiver operating characteristic curve and nomogram,and single sample gene set concentration analysis was used to study the correlation between the predictive model and tumor immune microenvironment in high-risk and low-risk groups.Results:We got a total of 5 m7G related lncRNA(MAFG-DT,AP003352.1,AC242842.1,AC024060.1,FAM111A-DT),which may be related to the prognosis of patients with bladder cancer.For predicting 1-,3-and 5-year survival rates,the area under the receiver operating characteristic curve was 0.757 and 0.722 and 0.739,respectively.Kaplan-Meier analysis showed that the prognosis of bladder cancer patients in high risk group was worse than that in low risk group.Immunoassay showed that the immune function of patients with bladder cancer in high risk group was more active.Conclusion:Prognostic markers based on m7G-related lncRNAs can be used to independently predict the prognosis of patients with bladder cancer and provide therapeutic targets for future clinical treatment.

分级加载条件下深部灰岩蠕应变特性研究

为研究深部灰岩蠕应变特性,采用GDS-VIS三轴流变仪对不同深度试样做分级加载单轴蠕变试验。基于试验结果,计算出灰岩长期强度与瞬时强度之比的均值为0.758,且深度越深比值越小;分析不同深度灰岩在各分级应力水平下轴向蠕应变增量与加载级数的变化规律,并将蠕应变增量进行一次累加,得到轴向蠕应变。结果表明:蠕应变增量随级数的增加呈现先减小后剧增的变化规律,且随深度增加,蠕应变增量达到谷底所对应的应力百分比有明显增大趋势;在分级加载过程中,随级数增加,轴向蠕应变先后呈现减速增加、等速增加和加速增加3个发展阶段;当恒载应力百分比为60%~70%时,蠕应变趋于稳定,当超过该范围后,蠕应变加速增加直至发生蠕变破坏。研究得到的这一应力百分比范围可以为识别深部灰岩是否会发生蠕变破坏提供依据。

长链非编码RNA HEIH在胃癌患者的表达及临床意义

目的探索肝细胞肝癌高表达(HEIH)长链非编码RNA(lncRNA)在胃癌患者的表达及临床意义。方法采用实时定量PCR检测50例胃癌患者的胃癌组织、癌旁组织及血浆标本lncRNA HEIH表达量,取50例健康对照者血浆标本作为对照。分析胃癌组织标本中lncRNA HEIH表达量与其临床病理特征的关系,绘制ROC曲线评估血浆lncRNA HEIH表达水平对胃癌的诊断效能。结果胃癌组织中lncRNA HEIH相对表达量高于癌旁组织(2.38±0.14 vs.1.59±0.09)(P<0.01),胃癌患者血浆中lncRNA HEIH相对表达量高于健康对照者(2.19±0.13 vs. 1.37±0.09)(P<0.01)。50例胃癌组织中,lncRNA HEIH相对表达量在肿瘤大小及有无淋巴结转移和远处转移组间比较均有统计学差异(P<0.05)。ROC曲线下面积为0.772;最佳诊断界值为1.570时,血浆lncRNA HEIH诊断胃癌的灵敏度和特异度分别为70.00%和80.00%。结论 lncRNA HEIH在胃癌患者的胃癌组织及血浆中高表达,有可能成为胃癌诊断及预后判断的潜在肿瘤标志物。