小鼠肝癌细胞特异性淋巴道转移与其基质金属蛋白酶分泌的关系

Zymographic analysis of matrix metalloproteinases in mouse carcinoma cell lines with different lymphatic metastasis potential

目的 探讨肿瘤细胞基质金属蛋白酶 (MMPs)分泌与特异性淋巴结转移的关系。方法 将自建高、低转移小鼠肝癌细胞系HCa F(高转移 )和HCa P(低转移 )在培养时 ,分别加入淋巴结、肝脏或脾脏匀浆 ,离心收集培养上清 ,用明胶酶谱法检测两株癌细胞在不同培养环境下分泌的MMPs酶谱 ,分析两者的差异。结果 在单纯RPMI16 40培养基中HCa F和HCa P细胞仅能分泌少量MMP 9(12 5 6± 15 7,2 6 42± 385 ) ,加入淋巴结匀浆后HCa F细胞分泌MMP 9显著增强 (12 40 3± 894) ,且分泌MMP 9的活性型和MMP 2 ;HCa P细胞也分泌MMP 9(90 6 8± 6 86 )及其活性型 (3 914± 12 5 3)和MMP 2(2 997± 1990 ) ,但量均明显低于HCa F细胞 ;加入肝脏或脾脏匀浆后 ,HCa F和HCa P细胞均不分泌MMPs。结论 HCa F细胞强于HCa P细胞的转移能力可能与HCa F细胞分泌MMPs的能力强于HCa P细胞相关 ,而HCa F和HCa P细胞仅在淋巴结成分诱导下才能分泌较多的MMPs,这可能与它们具有特异性地向淋巴结转移的功能密切相关。

Objective To investigate the correlation between lymphatic metastasis and matrix metalloproteinases (MMPs) activity. Methods Zymographic analysis was used to detect the difference of MMPs product and activities between high lymphatic metastatic cell line (HCa-F) and low lymphatic metastatic cell line (HCa-P), that were cultured in the medium including homogenate of lymph node or liver or spleen tissue. Results HCa-F and HCa-P cells produced small quantities of MMP-9 in 1640 medium. After adding lymph node homogenate to the culture medium, F cells produced a large quantity of MMP-9 as well as active MMP-9 and MMP-2. P cells also produced MMP-9、active MMP-9 and MMP-2, but were much lower than that of F cells. Neither F cells nor P cells produced MMPs in the media with homogenate of liver or spleen added. Conclusion The metastatic potential of F cells is stronger than that of P cells may be due to F cell's ability to secrete MMPs is stronger than P cell's, therefore the production of MMPs of hepatocarcinoma cells is closely correlated to their metastatic potential.