CAJ | 학술논문

目的：观察异硫氰酸苯乙酯(PEITC)对结肠癌细胞基质金属蛋白酶9(MMP-9)表达的影响及其分子机制。方法体外培养结肠癌细胞系SW480,分别用10、30、50μmol/L PEITC作用24 h,划痕愈合实验和侵袭实验分别检测PEITC对SW480细胞迁移与侵袭的影响。 RT-PCR检测MMP-9 mRNA的表达；Western blot检测MMP-9蛋白表达和蛋白激酶B( Akt)磷酸化。荧光素酶报告基因分析核转录因子κB( NF-κB)和MMP-9的活性。结果划痕愈合实验和Transwell小室实验结果显示,PEITC能在不影响细胞活性的条件下对SW480细胞侵袭和迁移具有一定抑制作用。同时,Western blot和RT-PCR结果证实PEITC能抑制MMP-9蛋白及mRNA表达。 PEITC能抑制Akt磷酸化以及NF-κB的转录活性。 PI3K抑制剂LY294002处理后,NF-κB-luc活性以及MMP-9蛋白表达显著降低。此外,不同浓度PEITC处理能显著抑制MMP-9的启动子活性。结论 PEITC是一种潜在的抗结肠癌细胞转移药物,它可能通过影响PI3K/NF-κB通路抑制MMP-9表达而发挥作用。
목적：관찰이류청산분을지(PEITC)대결장암세포기질금속단백매9(MMP-9)표체적영향급기분자궤제。방법체외배양결장암세포계SW480,분별용10、30、50μmol/L PEITC작용24 h,화흔유합실험화침습실험분별검측PEITC대SW480세포천이여침습적영향。 RT-PCR검측MMP-9 mRNA적표체；Western blot검측MMP-9단백표체화단백격매B( Akt)린산화。형광소매보고기인분석핵전록인자κB( NF-κB)화MMP-9적활성。결과화흔유합실험화Transwell소실실험결과현시,PEITC능재불영향세포활성적조건하대SW480세포침습화천이구유일정억제작용。동시,Western blot화RT-PCR결과증실PEITC능억제MMP-9단백급mRNA표체。 PEITC능억제Akt린산화이급NF-κB적전록활성。 PI3K억제제LY294002처리후,NF-κB-luc활성이급MMP-9단백표체현저강저。차외,불동농도PEITC처리능현저억제MMP-9적계동자활성。결론 PEITC시일충잠재적항결장암세포전이약물,타가능통과영향PI3K/NF-κB통로억제MMP-9표체이발휘작용。
Objective To investigate the effect and molecular mechanism of PEITC on matrix metalloproteinase-9 ( MMP-9) expression in human colon cancer SW480 cells. Methods Human colon cancer SW480 cell line was cultured in vitro,and treated with 10,30,50μmol/L PEITC for 24h. Cell viability,cell migration and invasion was analyzed by MTT assay, scratch healing assay and transwell membrane assay,respectably. Expression of MMP-9 mRNA was analyzed by RT-PCR. Pro-tein expression and phosphorylation of Akt were detected by Western blot. Activity of NF-κB and MMP-9 were analyzed by lu-ciferase reporter assay. Results PEITC reduced SW480 cells invasion and migration without affecting the viability of cells, as demonstrated by scratch healing and transwell membrane assay. Furthermore, Western blot and RT-PCR showed PEITC markedly inhibited MMP-9 activity and expression at both protein and mRNA levels. PEITC attenuated Akt phosphorylation and activity of NF-κB. Moreover,LY294002,an inhibitor of PI3K,significantly inhibited NF-κB transcriptional activity and MMP-9 expression. In addition,different concentration of PEITC could inhibit the promotion activity of MMP-9. Conclusion PEITC as a potential anti-invasive agent by inhibiting MMP-9 is involved in PI3K/NF-κB pathways.