FHL1通过阻滞细胞周期抑制舌鳞癌细胞的生长

发布时间：2018-10-22 12:49

【摘要】：研究背景: FHL（four and a half LIM domains）家族由4个半LIM结构域组成。2个半胱氨酸组成的双锌指模体（motif）是LIM结构域特有的结构，具有特异性，通过该结构与其他蛋白产生相互作用。到目前为止，FHL家族成员共发现五个成员，由FHL1-4和ACT（activator of CREM in testis）组成。FHL家族比较保守，五个成员除FHL4外，均在小鼠体内发现了相应的同源基因。FHL家族蛋白成员间的氨基酸序列具有较高的同源性，大约在40%-60%左右。FHL家族在肿瘤中扮演抑癌基因的角色，，但是FHL家族在舌癌中的功能至今没有详细的报道。目的:临床标本分析FHL1在舌癌组织及癌旁组织分布，FHL1过表达及低表达对舌癌细胞（Tca8113）生长的影响。方法:免疫组化法分析对比舌癌组织及癌旁组织中FHL1的分布，构建敲低FHL1的稳定克隆细胞株，通过流式细胞术、western blot、软琼脂实验、划痕实验、结晶紫实验、RT-PCR实验、裸鼠成瘤实验等实验观察FHL1对舌癌细胞（Tca8113）的影响。结果:①FHL1在舌癌癌旁组织中分布明显高于癌组织中。 ②体外条件下FHL1对舌癌细胞（Tca8113）不具有促进细胞迁移和侵袭的能力，能抑制舌癌细胞（Tca8113）锚着依赖性和锚着非依赖性生长。 ③体外条件下FHL1能阻滞细胞周期，并改变细胞周期相关调节蛋白CyclinA、Cyclin B、CyclinD、CyclinE蛋白的表达水平。 ④裸鼠成瘤实验结果显示FHL1具有抑制肿瘤细胞生长的作用。结论: FHL1在舌癌癌旁组织中分布明显高于癌组织中，体外条件下FHL1通过阻滞细胞周期，改变细胞周期相关调节蛋白的表达水平进而抑制细胞生长。
[Abstract]:Background the: FHL (four and a half LIM domains) family consists of four semi-LIM domains. Two cysteine dizinc finger motifs, (motif), are unique to the LIM domain and have specificity, through which they interact with other proteins. So far, five members of the FHL family have been found to be composed of FHL1-4 and ACT (activator of CREM in testis). The FHL family is conservative. The amino acid sequences among members of FHL family have high homology, about 40% to 60%. FHL family plays the role of tumor suppressor gene in tumor. But the role of the FHL family in tongue cancer has not been reported in detail. Aim: to analyze the distribution of FHL1 in tongue cancer tissues and adjacent tissues, and the effect of overexpression and low expression of FHL1 on the growth of tongue cancer cells (Tca8113). Methods: the distribution of FHL1 in tongue cancer tissues and adjacent tissues was analyzed by immunohistochemical method. Stable clone cell lines with low FHL1 knockout were constructed. Flow cytometry (FCM), western blot, soft Agar test, scratch test, crystal violet test, RT-PCR test were used. The effect of FHL1 on tongue cancer cell (Tca8113) was observed in nude mice. Results: the distribution of 1FHL1 in the adjacent tissues of tongue cancer was significantly higher than that in cancer tissues. 2 in vitro, FHL1 could not promote the migration and invasion of tongue cancer cells (Tca8113). It can inhibit anchor-dependent and anchor-independent growth of tongue cancer cells (Tca8113). 3 FHL1 can block cell cycle in vitro. The expression level of Cyclin D cyclin E protein was changed. 4 the results of nude mice tumorigenesis showed that FHL1 could inhibit the growth of tumor cells. Conclusion: the distribution of FHL1 in the adjacent tissues of tongue cancer is significantly higher than that in cancer tissues. In vitro, FHL1 inhibits cell growth by blocking cell cycle and changing the expression level of cell cycle related regulatory proteins.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.86

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