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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。
The mechanisms of anti-cancer metastasis of laminin glycopeptides (LN GPs) have been studied in terms of proliferation, apoptosis, and secretion of matrix metalloproteinases (MMPs) from cancer cells. METHODS: The human hepatoma cell line Bel740 2 cells were incubated on the laminin matrix for a certain period of time. The total number of viable cell populations was measured by thiazolyl blue colorimetry. 3H TdR assay was used to measure the DNA synthesis of cancer cells; fluorescein-activated cell sorting (FACS) was used to determine the cell cycle; cells were counted by Giemsa and counted for mitotic index; FACS and acridine orange staining were used to detect apoptosis. . Gelatin zymography was used to detect the level of MMPs secreted by cancer cells in serum-free culture supernatant. The glycopeptide group was supplemented with LN GPs (50 mg·L-1). RESULTS: After incubation with cancer cells, the total number of viable cell populations increased, 3H TdR incorporation increased, G1 phase cells decreased, and S phase cells increased. On the contrary, the total number of living cell populations in the LN GPs group was significantly lower, 3H TdR incorporation was decreased, G1 phase cells were increased and S phase cells were decreased, similar to the results in the uncoated group. However, no obvious cancer cell apoptosis was detected by either method. In the LN GPs group, the secretion and activation of MMPs in human hepatocellular carcinoma cell line Bel740 2 cells and high metastatic potential human giant cell lung cancer cell line PG cells were significantly reduced. Conclusion: LN can promote the proliferation of cancer cells, while LN GPs can inhibit this effect and can significantly inhibit the secretion of MMPs in cancer cells, which may be an important part of its inhibition of cancer cell invasion and metastasis.