Matriptase表达下调对胰腺癌细胞株SW1990侵袭的影响

Effects of matriptase down regulation on invasion of human pancreatic cancer cells SW1990

目的探讨matriptase基因沉默对胰腺癌细胞株SW1990侵袭力的影响。方法采用脂质体法将不同浓度的靶向matriptase的siRNA（Ma—siRNA）转染人胰腺癌SW1990细胞株，以转染阴性对照的siRNA（NC—siRNA）作为对照组。采用荧光定量PCR法和蛋白质印迹法检测各组细胞matriptase mRNA和蛋白的表达水平，采用Transwell小室检测细胞侵袭能力，明胶酶谱法检测细胞matriptase及MMP-9的活性。结果NC-siRNA组及12．5、25、50nmoL／L Ma—siRNA组SW1990细胞matriptase mRNA相对表达量分别为1．000、0．417±0．006、0．233±0．068、0．221±0．092，蛋白表达量分别为0．736±0．066、0．498±0．036、0．341±0．118、0．239±0．050，Ma－siRNA各组细胞matriptase mRNA及蛋白表达量均显著低于NC—siRNA组，差异具有统计学意义（P值均〈0．01）；matriptase活性分别为1．501±0．165、1．211±0．265、0．645±0．165、0．620±0．003，MMP-9活性分另0为0．929±0．260、0．484±0．364、0．352±0．113、0．346±0．121，其中25、50nmol／L Ma—siRNA组细胞的matriptase及MMP-9活性较NC—siRNA组显著下降，差异有统计学意义（P值〈0．05或〈0．01）。NC-siRNA组的穿膜细胞数为（256±1）个／200倍视野，25nmol／L Ma—siRNA组为（109±3）个／200倍视野，25nmol／L Ma—siRNA组细胞侵袭力较对照组下降（57．4±5．4）％。结论抑制matriptase基因表达可降低胰腺癌SW1990细胞的侵袭能力，其机制可能与matriptase及MMP-9酶活性下降有关。

Objective To investigate the effect of the down regulation of matriptase expression on invasion of human pancreatic cancers cells SW1990. Methods Small interfering RNA targeting at matriptase （Ma-siRNA） was transfected into human pancreatic cancers SW1990 cells, and nonsense siRNA （NC-siRNA） group was used as control. Real time PCR assay and Western blot were used to detect the expression of matriptase mRNA and protein. Transwell assay was used to examine the invasion ability of cancer cells. The enzymatic activity of matriptase and MMP-9 was determined by gelatin zymography assay. Results The expression level of matriptase mRNA in NC-siRNA group, 12.5, 25, 50 nmol/L Ma-siRNA group were 1.000, 0.417± 0.006, 0.233 ± 0.068, 0.221 ± 0.092; and the protein expression of matriptase were 0.736 ±0.066, 0. 498 ± 0. 036, 0. 341 ± 0.118, 0. 239 ± 0. 050, respectively. The matriptase mRNA and protein expression in Ma-siRNA groups was significantly lower than those in NC-siRNA group, and the difference between the two groups was statistically significant （ P 〈 0.05 ）. The enzymatic activity of matriptase were 1. 501 ±0. 165, 1. 211 ±0.265, 0. 645 ±0. 165, 0. 620 ±0. 003, and the enzymatic activity of MMP-9 were 0.929 ± 0.260, 0.484 ± 0.364, 0.352 ± 0. 113, 0.346 ± 0. 121, and the enzymatic activity of matriptase and MMP-9 in 25, 50 nmol/L Ma-siRNA groups was significantly lower than that in NC-siRNA group, and the difference was statistically significant （ P 〈0.05 or 〈0.01 ）. The number of transmembrane cell was （256 ± 1 ）/per 200 power field, and it was （ 109 ± 3 ）/per 200 power field in 25 nmol/L Ma-siRNA group, and the invasion ability of the cells in 25 nmol/L Ma-siRNA group was decreased by （ 57.4 ± 5.4 ） % when compared with that of control group. Conclusions Down-regulation of matriptase inhibits invasion ability of pancreatic cancer SW1990 cells, and this result may be due to the down regulated enzymatic activity of matriptase and MMP-9.