粉防己碱对喉鳞状细胞癌抑癌基因Brg1表达的影响及相关机制的研究

发布时间：2018-03-08 16:08

  本文选题：喉癌　切入点：Brg1　出处：《吉林大学》2010年博士论文　论文类型：学位论文

【摘要】： 本研究通过分析喉鳞癌组织中Brg1蛋白的表达,粉防己碱(Tet)对喉鳞状细胞癌细胞系中Brg1及相关基因表达的影响,探讨Brg1基因与喉鳞癌发生发展的关系,以及Tet的抗肿瘤机制。采用免疫组化方法分析Brg1在喉鳞癌组织中的表达,比较其在正常喉粘膜、声带息肉、喉癌前病变及喉鳞状细胞癌中的表达情况。采用免疫荧光法检测Tet对Hep-2细胞中Brg1表达的影响,采用MTT法检测不同浓度的Tet对喉鳞癌Hep-2细胞增殖的影响,采用流式细胞仪检测不同浓度Tet对细胞周期的影响,以及应用fluo-3检测Tet对细胞内钙离子浓度的影响,表明在喉鳞状细胞癌中Brg1表达下降,在喉癌发展过程中有重要作用。Tet可使Hep-2细胞中Brg1表达增强,抑制肿瘤细胞生长,并可使Hep-2细胞内钙离子浓度降低。采用免疫荧光法检测Tet对Hep-2细胞中受Brg1调节的钙调蛋白AHNAK的表达,并进一步应用实时RT-PCR方法检测细胞内Brg1和AHNAK mRNA水平,结果发现Tet可使Hep-2细胞内Brg1和AHNAK转录增强。表明Tet可使受Brg1调控的钙调蛋白AHNAK的表达增强,Brg1的表达可能与细胞内钙相关。Brg1作为一种抑癌基因在喉鳞状细胞癌中表达下降,可能与喉癌的发生密切相关。Tet为一中药抗肿瘤药物,可使Hep-2细胞中Brg1表达增强,其抗肿瘤作用可能通过影响Brg1表达而实现。
[Abstract]:By analyzing the expression of Brg1 protein in laryngeal squamous cell carcinoma and the effect of Tetrandrine on the expression of Brg1 and related genes in laryngeal squamous cell carcinoma cell line, the relationship between Brg1 gene and the occurrence and development of laryngeal squamous cell carcinoma was studied. Immunohistochemical method was used to analyze the expression of Brg1 in laryngeal squamous cell carcinoma (LSCC), and to compare the expression of Brg1 in normal laryngeal mucosa and vocal cord polyp. The effect of Tet on the expression of Brg1 in Hep-2 cells was detected by immunofluorescence assay, and the effect of different concentrations of Tet on the proliferation of Hep-2 cells in laryngeal squamous cell carcinoma (LSCC) was detected by MTT assay. Flow cytometry was used to detect the effect of different concentrations of Tet on cell cycle, and fluo-3 was used to detect the effect of Tet on intracellular calcium concentration, which indicated that the expression of Brg1 decreased in laryngeal squamous cell carcinoma. Tet plays an important role in the development of laryngeal carcinoma. Tet can enhance the expression of Brg1 in Hep-2 cells and inhibit the growth of tumor cells. Immunofluorescence assay was used to detect the expression of calmodulin AHNAK regulated by Brg1 in Hep-2 cells by Tet, and the levels of Brg1 and AHNAK mRNA in Hep-2 cells were detected by real-time RT-PCR method. The results showed that Tet could enhance the expression of Brg1 and AHNAK in Hep-2 cells, suggesting that Tet could enhance the expression of Brg1 regulated calmodulin AHNAK and the expression of Brg1 might be related to the decrease of intracellular Ca2 + related. Brg1 as a tumor suppressor gene in laryngeal squamous cell carcinoma. It may be closely related to the occurrence of laryngeal carcinoma. Tet is a traditional Chinese medicine anti-tumor drug, which can enhance the expression of Brg1 in Hep-2 cells, and its anti-tumor effect may be realized by affecting the expression of Brg1.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R739.65

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