Matriptase与子宫内膜癌细胞株侵袭迁移能力的相关性研究

发布时间：2018-09-06 17:17

【摘要】：目的:探讨Matriptase与子宫内膜癌细胞侵袭转移能力的相关性。方法:采用荧光定量PCR和Western blot法检测HEC-1A、HEC-1B和RL952细胞中Matriptase、HAI-1 mRNA和蛋白表达水平;构建Matriptase基因SiRNA慢病毒载体,采用RNA干扰技术,对Matriptase高表达的HEC-1A和RL952细胞进行瞬时转染,从而达到抑制Matriptase基因表达水平的目的。应用细胞划痕实验及穿膜小室实验观察干扰前后癌细胞体外侵袭及迁移能力的改变。结果:荧光定量PCR检测结果显示,Matriptase和HAI-1 mRNA在HEC-1A细胞(0.2123±0.021和0.2827±0.049)和RL-952细胞(0.1778±0.013和0.2420±0.032)中为均阳性表达;在HEC-1B细胞中为阴性表达(0.000695±0.00012和0.00263±0.00043)。Western blot结果显示,Matriptase和HAI-1在HEC-1A细胞和RL-952细胞中为阳性表达,在HEC-1B细胞中为阴性表达。转染siRNA后,HEC-1A细胞和RL-952细胞的Matriptase mRNA和蛋白表达水平显著下降,抑制率达80%以上,HAI-1表达水平未见明显改变,Matriptase/HAI-1比值明显降低(HEC-1A:0.75 vs 0.11,P0.01;RL-952:0.0.73 vs 0.12,P0.01)。细胞划痕实验和穿膜小室实验显示,癌细胞体外侵袭及迁移能力显著下降。结论:Matriptase表达水平与子宫内膜癌细胞侵袭迁移能力呈正相关,Matriptase/HAI-1比值降低亦可能抑制癌细胞的侵袭迁移。
[Abstract]:Objective: to investigate the relationship between Matriptase and invasion and metastasis of endometrial cancer cells. Methods: the expression levels of Matriptase,HAI-1 mRNA and protein in HEC-1A,HEC-1B and RL952 cells were detected by fluorescence quantitative PCR and Western blot, and the Matriptase gene SiRNA lentivirus vector was constructed. HEC-1A and RL952 cells with high expression of Matriptase were transfected by RNA interference technique. In order to achieve the purpose of inhibiting the expression level of Matriptase gene. Cell scratch test and transmembrane chamber test were used to observe the invasion and migration ability of cancer cells before and after interference. Results: the results of fluorescence quantitative PCR showed that both matriptase and HAI-1 mRNA were positive in HEC-1A cells (0.2123 卤0.021 and 0.2827 卤0.049) and RL-952 cells (0.1778 卤0.013 and 0.2420 卤0.032), and negative in HEC-1B cells (0.000695 卤0.00012 and 0.00263 卤0.00043). Western blot showed that matriptase and HAI-1 were positive in HEC-1A cells and RL-952 cells. Negative expression was found in HEC-1B cells. After transfection of siRNA, the expression of Matriptase mRNA and protein in HEC-1A cells and RL-952 cells decreased significantly, and the inhibition rate was more than 80%. There was no significant change in the ratio of Matriptase-1 / HAI-1 (HEC-1A:0.75 vs 0.11p0.01RL-952: 0.73 vs 0.12P0.01). Cell scratch assay and transmembrane chamber test showed that the ability of invasion and migration of cancer cells in vitro decreased significantly. Conclusion there is a positive correlation between the expression of 1: Matriptase and the ability of invasion and migration of endometrial cancer cells. The decrease of the ratio of Matriptase- / HAI-1 may also inhibit the invasion and migration of endometrial cancer cells.
【作者单位】：福建医科大学教学医院福建省妇幼保健院妇科肿瘤实验室;福建医科大学附属协和医院妇产科;
【基金】：福建省自然科学基金(No:2012J01310)
【分类号】：R737.33

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