lncRNA/microRNA-29家族调控瘢痕疙瘩增殖侵袭及纤维化的作用机制

The performance characteristics of keloid is known as excessive invasion，proliferation, fibrosis, and high recurrence rate after surgery alone therapy. Keloid is hard to be diagnosed from hypertrophic scar in early stages. Nowadays, the specific mechanism is not clear. And there are no safe and effective clinical treatment strategies as well. Studies have shown that, lncRNA and miR-29 family have played an important role in the inhibition of invasion, proliferation and tissue fibrosis diseases, and lots of researches been focused on this valuable area in recent years. Our previous studies have suggested that miR-29c-5p (one of the membermiR-29 family) have a function of regulation of keloid fibroblast proliferation and fibrosis. Based on our preliminary studies and current situation of the research, this project intends to systematic study the expression characteristics of lncRNAs and miR-29 family in keloids and hypertrophic scars through bioinformatics, looking for the key target genes of regulation of keloid and the identification biomarker between keloid and hypertrophic scar, investigating the relationship between lncRNA/miR-29 family and the signaling pathways relative to invasion, proliferation and fibrosis, verifying the mechanism of keloid proliferation, invasion and fibrosis regulated by lncRNA/miR-29 family, finding out the effective clinical treatment intervention targets, further more providing the theory foundation for effective treatment of keloid.

瘢痕疙瘩表现为过度增殖侵袭和纤维化的特性，早期与增生性瘢痕极难鉴别，单纯手术等治疗后复发率极高，现机制未明，临床尚无安全有效的治疗手段。研究表明，lncRNA与miRNAs相互调控在多种增殖侵袭及纤维化疾病的发生发展中起重要作用，是极具研究价值的热点。我们前期研究提示，miR-29家族中miR-29c-5p可调控瘢痕疙瘩组织增殖及纤维化作用。结合前期基础及研究现状，本项目拟结合生物信息学，对瘢痕疙瘩及增生性瘢痕中lncRNA/miR-29家族的表达特点进行系统研究，寻找瘢痕疙瘩与增生性瘢痕鉴别的分子标志调控瘢痕疙瘩的核心靶基因，并通过调控关键基因的表达后，探索lncRNA/miR-29家族与瘢痕疙瘩增殖侵袭和纤维化相关的信号通路系统的关系，探明lncRNA/miR-29家族调控瘢痕疙瘩增殖侵袭及纤维化的作用机制，为瘢痕疙瘩的临床治疗寻找到有效的干预靶点进而为有效治疗瘢痕疙瘩奠定基础。

瘢痕疙瘩表现为过度增殖侵袭和纤维化的特性，早期与增生性瘢痕极难鉴别，单纯手术等治疗后复发率极高，现机制未明，临床尚无安全有效的治疗手段。研究表明，lncRNA与miRNAs 相互调控在多种增殖侵袭及纤维化疾病的发生发展中起重要作用，是极具研究价值的热点。.课题组前期研究提示，miR-29家族中miR-29c-5p可调控瘢痕疙瘩组织增殖及纤维化作用。通过结合前期基础及研究现状，通过提取标本库中冻存的组织，提取RNA行RT-PCR检测miR-29家族在瘢痕疙瘩、增生性瘢痕及正常皮肤组织中的差异性表达情况。同时，课题组提取新鲜组织标本送全转录组测序及miRNA芯片测序，在组织水平检测瘢痕疙瘩、增生性瘢痕、正常皮肤mRNA、lncRNA、miRNA表达谱的差异性表达情况，并行生物信息学分析，探索病理性瘢痕发生发展过程中miRNA-29家族与lncRNA的相互作用模式。. 通过研究，在组织水平上验证了miR-29家族在瘢痕疙瘩、增生性瘢痕、正常皮肤中的差异性表达情况。通过全转录组测序、miRNA芯片测序，在分子层面探索了病理性瘢痕发生发展的机制。进一步通过生物信息学分析探索了miRNA-29家族与lncRNA的互作模式。发现瘢痕疙瘩组与增生性瘢痕组间筛选出3个差异表达miRNA，包括1个上调miRNA和2个下调miRNA；瘢痕疙瘩组与正常皮肤组间筛选出139个差异表达miRNA，包括63个上调miRNA和76个下调miRNA；增生性瘢痕组与正常皮肤组间筛选出150个差异表达miRNA，包括67个上调miRNA，83个下调miRNA。其中miR-188-5p在增生性瘢痕的表达约为瘢痕疙瘩中的10倍，是增生性瘢痕和瘢痕疙瘩的重要差异表达miRNA。. 此外，本研究还在原有的计划上进行了拓展，分别深入挖掘了miRNA芯片数据及lncRNA测序数据，发现了在病理性瘢痕发生发展过程中起重要作用的miRNA（miRNA-188-5p）及lncRNA（MIR99AHG:3，LNC-DHX32-3:1，LNC-PRKACB-2:1，LNC-STARD4-D:1，LNC-DLK-35:125，LNC-ADPRM）。. 因此，本研究从多个维度（mRNA、lncRNA、miRNA）对病理性瘢痕的发生发展机制进行了深入的挖掘，为将来的病理性瘢痕的研究奠定了夯实的基础