Matriptase与子宫内膜癌细胞株侵袭迁移能力的相关性研究

The relationship between Matriptase expression and the invasion and migration of endometrial cancer cells

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,P<0.01;RL-952:0.0.73 vs 0.12,P<0.01)。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。

Objective：To explore the relationship between Matriptase expression and the invasion and migration of endometrial cancer cells. Methods：Detect three endometrial cancer cells mRNA and protein expression level of Matriptase,and HAI-1 by fluorescence quantitative PCR and Western blot. We successfully constructed Matriptase lentivirus siRNA plasmid expression vectors and apply the transient transfection technique in order to suppress the expression of Matriptase of HEC-1A and RL952 which express high level of Matriptase. Then,the ability of invasion and migration were compared before and after transfection using scratch assay and transwell chamber assay separately. Results： The mRNA expressions of Matriptase and HAI-1 were positive in HEC-1A cells（0. 2123 ± 0. 021,0. 2827 ± 0. 049） and RL-952 cells（0. 1778 ± 0. 013,0. 2420 ± 0. 032）,while they were negative in HEC-1B cells（0. 000695 ±0. 00012,0. 00263 ± 0. 00043）. A similay result was observed in the protein expression of Matriptase and HAI-1. The mRNA and protein expression level of Matriptase were significantlydecreased as much as 80% after transfection with siRNA target on Matriptase. The ration of Matriptase / HAI-1 was derecesed from 0. 75 to 0. 11 in HEC-1A cells and from 0. 73 to 0. 12 in RL-952 cells（ P〈0. 01）. The scratch assay and millicell chamber invasion assay showed that the ability of invasion and migration were both marked suppressed. Conclusion： The invasion and migration of endometrial cancer cells was positive related with the Matriptase expression,so was it correlated with the Matriptase / HAI-1 ratio.