CAJ | 학술논문

目的:研究小檗碱对高转移性人巨细胞肺癌PG细胞侵袭和迁移的作用并探讨其作用机制.方法:采用琼脂糖滴法观察PG细胞的运动能力,用下室加FN的Transwell小室法观察PG细胞的趋化运动能力,用铺Martrigel的Transwell小室观察PG细胞的侵袭能力.通过SABC免疫细胞化学法及图像分析软件,半定量PG细胞MMP2、TIMP2的表达;RT-PCR法检测PG细胞MMP2 mRNA和TIMP2 mRNA的表达.结果:小檗碱(10 mg/L)处理PG细胞24 h后:(1)PG细胞的迁移距离明显比对照PG短(P＜0.01),趋向FN迁移的穿膜细胞数明显少于对照组(P＜0.01);(2)PG细胞与FN、Martrigel的黏附减少,与对照组比有显著差异(P＜0.01);(3)穿过铺Martrigel的Transwell小室微孔滤膜的PG细胞数明显减少,即可抑制PG细胞的侵袭能力(P＜0.05);(4)MMP2表达明显减少(P＜0.05),而TIMP-2的表达有升高趋势,与对照比无显著差异,但MMP2/TIMP2比值降低;(5)小檗碱使PG细胞TIMP2 mRNA表达明显增加(P＜0.05),而对MMP2 mRNA表达无影响,MMP2 mRNA/TIMP2 mRNA比值降低.结论:抑制肿瘤细胞的运动能力、与细胞外基质的黏附、对细胞外基质侵袭,调节MMP2/TIMP2表达的平衡,从而维持细胞外基质的完整性,可能是小檗碱抗侵袭和转移作用的机制之一.
목적:연구소벽감대고전이성인거세포폐암PG세포침습화천이적작용병탐토기작용궤제.방법:채용경지당적법관찰PG세포적운동능력,용하실가FN적Transwell소실법관찰PG세포적추화운동능력,용포Martrigel적Transwell소실관찰PG세포적침습능력.통과SABC면역세포화학법급도상분석연건,반정량PG세포MMP2、TIMP2적표체;RT-PCR법검측PG세포MMP2 mRNA화TIMP2 mRNA적표체.결과:소벽감(10 mg/L)처리PG세포24 h후:(1)PG세포적천이거리명현비대조PG단(P＜0.01),추향FN천이적천막세포수명현소우대조조(P＜0.01);(2)PG세포여FN、Martrigel적점부감소,여대조조비유현저차이(P＜0.01);(3)천과포Martrigel적Transwell소실미공려막적PG세포수명현감소,즉가억제PG세포적침습능력(P＜0.05);(4)MMP2표체명현감소(P＜0.05),이TIMP-2적표체유승고추세,여대조비무현저차이,단MMP2/TIMP2비치강저;(5)소벽감사PG세포TIMP2 mRNA표체명현증가(P＜0.05),이대MMP2 mRNA표체무영향,MMP2 mRNA/TIMP2 mRNA비치강저.결론:억제종류세포적운동능력、여세포외기질적점부、대세포외기질침습,조절MMP2/TIMP2표체적평형,종이유지세포외기질적완정성,가능시소벽감항침습화전이작용적궤제지일.
AIM: To study the effect of berberine(Ber) on invasion and migration of PG cells from a high metastatic human giant lung carcinoma cell line and to explore its mechanism. METHODS: Agarose drop method was used to detect PG migration; transwell cabin with FN in lower chamber was adopted to detect PG chemotaxis. PG adhesion to FN and martrigel was detected by MTT; PG invasive ability was determined by transwell cabin covered with martrigel. Expression of MMP2/TIMP2 protein and mRNA were detected by quantitative immunocytochemical method and RT - PCR respectively. RESULTS: After PG was treated by Ber( 10 mg/L) for 24 h: 1 ) migration distance of Ber- treated PG cells was markedly shorter than that of control cells (P ＜0. 01 ) and the number of passed membrane cells towards FN was much fewer than that of control cells ( P ＜ 0. 01 ); 2) PG adhesion to FN and martrigel was inhibited remarkably by Ber compared with control PG; 3) the migration of PG cells through the martrigel -coated transwell was significantly inhibited by the addition of Ber; 4) MMP2 expression was reduced significantly(P ＜0. 01 ), while the TIMP2 expression showed up - regulating tendency, but had no differences compared with control group (P ＞ 0. 05). The MMP2/TIMP2 ratio was decreased; 5 )the MMP2 mRNA/TIMP2 mRNA ratio was decreased by Ber. CONCLUSION: Inhibition of cell migration, adhesion to ECM and invasion into ECM of tumor cells and regulation of homeostasis between MMPs and TIMPs to maintain ECM integrity may be the basic mechanism of inhibitive effect of Ber on invasion and metastasis of tumors.