层粘连蛋白总糖肽抗癌细胞转移机制的研究  

Further studies on the anti metastasis mechanism of Laminin glycopeptides

作　　者：路艳艳[1] 周柔丽[1] 张莎[1] 蒋新农[1] 

机构地区：[1]北京大学基础医学院细胞生物学教研室,北京100083

出　　处：《北京医科大学学报》2000年第3期258-261,共4页Journal of Peking University(Health Sciences)

基　　金：国家自然科学基金 !(395 70 80 1)

摘　　要：从癌细胞增殖、凋亡和基质金属蛋白酶 (matrixmetalloproteinases,MMPs)的分泌等方面研究层粘连蛋白总糖肽 (laminin glycopeptides,LN GPs)抗癌细胞转移的机制。 方法 :将人肝癌细胞系Bel740 2细胞于层粘连蛋白基质上孵育一定时间后 ,噻唑蓝比色法测活细胞群体总数。3H TdR参入法测癌细胞DNA的合成 ;荧光素活化的细胞分拣术 (fluorescentactivatedcellsorting ,FACS)测定细胞周期 ;细胞经吉姆萨染色后计数有丝分裂指数 ;FACS和丫啶橙染色法检测细胞的凋亡。明胶酶谱分析法检测无血清培养上清中癌细胞分泌MMPs的水平。糖肽组加LN GPs (5 0mg·L-1)。结果 :癌细胞孵育后 ,活细胞群体总数增加 ,3H TdR的参入升高 ,G1期细胞减少而S期细胞增加。相反 ,加LN GPs组活细胞群体总数明显降低 ,3H TdR的参入下降 ,G1期细胞增加而S期细胞减少 ,与未包被组结果相近。但两种方法均未检测到明显的癌细胞凋亡。LN GPs组人肝癌细胞系Bel740 2细胞和高转移潜能的人巨细胞肺癌细胞系PG细胞的MMPs分泌和活化明显降低。结论 :LN可促进癌细胞的增殖 ,而LN GPs能抑制这一作用 ,并可明显抑制癌细胞MMPs的分泌 ,这可能是其抑制癌细胞侵袭和转移的重要环节。Objective: To further study the mechanism of anti metastasis effects of laminin glycopeptides (LN GPs) on carcinoma cell proliferation, apoptosis and secretion of matrix metalloproteinases (MMPs). Methods: Human hepatocellular carcinoma (HCC) cell line Bel7402 cells in serum free medium were incubated on laminin (LN) coated substrate for indicated time with or without LN GPs at the final concentration of 50 mg·L -1 . The total number of survival cells after incubating for 24, 48, 72 and 96 hours was assayed by MTT method. DNA synthesis of the incubated cells was analyzed by 3H TdR incorporation. Cell cycle was analyzed by FACS (fluorescent activated cell sorting, FACS). Mitotic index of Giemsa stained cells was counted. Carcinoma cell apoptosis was detected by both FACS and acridine orange (AO) staining methods. MMPs secretion of incubated cells was analyzed by gelatin zymography. Results: The total number of survival carcinoma cells incubated on LN was significantly larger than that on uncoated substrate. LN could promote HCC cell DNA synthesis as shown by 3H TdR incorporation, decrease the percentage of carcinoma cells in G 1 phase and increase the percentage of carcinoma cells in S phase as shown by FACS. In contrast, LN GPs could inhibit the effect of LN by inhibiting carcinoma cell 3H TdR incorporation, increasing the percentage of carcinoma cells in G 1 phase and decreasing the percentage of carcinoma cells in S phase. MMPs secretion of carcinoma cells on LN in the presence of LN GPs was much less compared with that of cells on LN alone. Conclusion: LN could stimulate carcinoma cell proliferation LN GPs could significantly inhibit the effect by inhibiting DNA synthesis and could inhibit the secretion of MMPs of carcinoma cells, which might contribute to their decreased invasive and metastatic phenotype.

关 键 词：层粘连蛋白 糖肽类 肿瘤转移 药物疗法 抗癌药 

分 类 号：R979.1[医药卫生—药品] R730.53[医药卫生—药学]