HnRNP L在口腔鳞癌中的作用及调控SRSF3的研究

发布时间：2018-04-10 16:14

本文选题：口腔鳞状细胞癌 + hnRNP　；参考：《武汉大学》2017年硕士论文

【摘要】：背景:口腔鳞状细胞癌(Oral squamous cell carcinoma,OSCC)是人类头颈部最常见的恶性肿瘤,具有高死亡率,是影响人类健康的重大疾病之一,但其发生、发展的机制并没有被完全研究清楚。核不均一核糖核蛋白(Heterogeneous nuclear ribonucleoprotein,hnRNP)是一个蛋白超家族,种类繁多,其中hnRNP L是一个具有多种功能的可变剪切调控因子,参与一系列与RNA有关的生物学过程,如染色质修饰、无内含子mRNA的出核、poly(A)位点的选择、维持mRNA的稳定等。目前越来越多的研究表明,hnRNP L与肿瘤的形成和发展都有着重要关联,然而对于hnRNP L与口腔鳞状细胞癌的关系仍未知。目的:本研究的目的是探索hnRNP L与口腔鳞状细胞癌的关系及其可能存在的调控机制。方法:1.应用免疫组织化学染色方法,检测hnRNP L在口腔鳞状细胞癌组织和正常口腔粘膜组织中的表达情况,并与SRSF3的表达水平进行相关性分析。2.应用转染技术降低或增强293细胞和CAL27细胞中hnRNP L或SRSF3的表达,通过免疫印迹实验检测hnRNP L和SRSF3的表达情况。3.应用RNAi技术降低CAL27细胞中hnRNP L的表达,通过逆转录PCR检测SRSF3的转录水平和外显子4的可变剪切。结果:1.HnRNP L在口腔鳞状细胞癌组织中的表达程度显著高于其在正常口腔粘膜组织中的表达程度,且hnRNP L与SRSF3在口腔鳞癌组织中的表达水平呈正相关的关系。2.降低或增强293细胞和CAL27细胞中hnRNP L的表达后,SRSF3的表达也降低或增强;降低293细胞和CAL27细胞中SRSF3的表达后,hnRNP L的表达也降低。3.抑制CAL27细胞中hnRNPL的表达后,SRSF3mRNA的整体转录水平降低,包含外显子4的剪切异构体的相对比例增高。结论:HnRNP L在口腔鳞状细胞癌组织中高表达,且与SRSF3在口腔鳞癌中的表达呈正相关。HnRNP L可能通过转录水平和转录后水平的共同作用来调控SRSF3的表达,hnRNP L能够调控SRSF3外显子4的可变剪切,SRSF3可能是hnRNP L的靶基因。
[Abstract]:Background: oral squamous cell carcinoma (OSCC) is the most common malignant tumor in human head and neck. It has a high mortality rate and is one of the major diseases affecting human health. However, the mechanism of its occurrence and development has not been fully studied.Nuclear heterogeneous nuclear ribonucleoprotein (RNPs) is a superfamily of proteins, in which hnRNP L is a variable shear regulator with many functions involved in a series of biological processes related to RNA, such as chromatin modification.The selection of nucleocapsid polyanus A (PPA) site in mRNA without intron, maintaining the stability of mRNA, etc.At present, more and more studies show that hnRNP RNP L is associated with tumor formation and development. However, the relationship between hnRNP L and oral squamous cell carcinoma is still unknown.Objective: to explore the relationship between hnRNP L and oral squamous cell carcinoma and its possible regulatory mechanism.Method 1: 1.Immunohistochemical staining was used to detect the expression of hnRNP L in oral squamous cell carcinoma (OSCC) and normal oral mucosa.The expression of hnRNP L or SRSF3 in 293 cells and CAL27 cells was decreased or enhanced by transfection technique. The expression of hnRNP L and SRSF3 was detected by Western blotting.RNAi technique was used to reduce the expression of hnRNP L in CAL27 cells. Transcriptional level of SRSF3 and variable shearing of exon 4 were detected by reverse transcription PCR.Results 1. The expression of HnRNP L in oral squamous cell carcinoma was significantly higher than that in normal oral mucosa, and there was a positive correlation between the expression of hnRNP L and SRSF3 in oral squamous cell carcinoma.After decreasing or enhancing the expression of hnRNP L in 293 cells and CAL27 cells, the expression of SRSF3 in 293 cells and CAL27 cells was also decreased or enhanced, and the expression of SRSF3 RNP L in 293 cells and CAL27 cells was also decreased.After inhibiting the expression of hnRNPL in CAL27 cells, the overall transcription level of SSRSF3 mRNA decreased, and the relative proportion of the shearing isomers containing exon 4 was increased.ConclusionThe high expression of% HnRNP L in oral squamous cell carcinoma is found.HnRNP L may regulate the expression of SRSF3 by the interaction of transcription level and posttranscriptional level. HnRNP L may be the target gene of hnRNP L by regulating the variable shearing of SRSF3 exon 4.
【学位授予单位】：武汉大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8

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