摘要
目的探讨大蒜素对结肠癌LoVo细胞增殖、侵袭和转移能力的影响，及其抗结肠癌侵袭转移作用的机制。方法采用MTT法检测大蒜素对LoVo细胞增殖抑制能力；肿瘤细胞体外运动实验、黏附实验、迁移实验和Transwell体外侵袭实验检测大蒜素对LoVo细胞运动、黏附、侵袭转移的抑制作用；实时荧光定量RT．PCR法检测大蒜素对LoVo细胞侵袭转移相关基因基质金属蛋白酶2（MMP-2）、基质金属蛋白酶抑制剂2（TIMP-2）、CDl47、血管内皮生长因子（VEGF）、转移抑制基因（nm23-H1）、乙酰肝素酶（HPA）、尿激酶纤维蛋白溶酶原激活剂受体（uPAR）mRNA表达的影响。结果大蒜素可明显抑制人结肠癌LoVo细胞增殖，具有时问剂量依赖性；“无毒”浓度（3和6μg／m1）大蒜素能够抑制LoVo细胞的体外运动、黏附、侵袭转移能力，作用24h后对LoVo细胞趋化运动抑制率分别为24％、50％（t=4．543、12．348，P=0．010、0．001），黏附抑制率分别为19％、28％（t=6．145、6．355，P=0．004、0．003），迁徙抑制率分别为28％、46％（t=8．065、28．435，均P〈0．01），侵袭抑制率分别为44％、65％（t=21．274、26．288，均P〈0．01）；3和6μg/ml浓度大蒜素能够下调VEGF、uPAR及HPAmRNA的表达（t=7．129、6．764、8．497，P=0．002、0．002、0．001），对TIMP-2，CD147，nm23-H1mRNA表达无明显影响（t=0．341、1．889、0．914，P=0．059、0．132、0．412）；LoVo细胞不表达MMP-2mRNA。结论“无毒”浓度（3和6μg/ml）大蒜素对LoVo细胞的迁移运动、黏附、侵袭转移能力有明显抑制作用，该作用可能与抑制VEGF、uPAR及HPAmRNA的表达有关。
Objective To study the effect of allicin on invasion and metastasis of human colon cancer cell line LoVo in vitro and furthermore elucidate its antieaneer mechanisms. Methods MTT assay was used to test dynamically the effect of cell growth inhibition. The inhibitory effects of alliein on movement, adhesiveness and invasiveness of LoVo cells were evaluated by the migratory test, adhesion test and Transwell chamber experiment. Quantitative real-time reverse transcription PCR （real-time RT-PCR ） was performed to quantify the mRNA expression of MMP-2, TIMP-2, CD147, VEGF, nm23-H1, HPA and uPAR. Results Allicin had inhibitive effects on growth of LoVo cells in a dose and time-dependent manner. Alliein at non-cytotoxic concentration （3 and 6 μg/ml） could obviously suppress the movement, adhesion and invasive capability of LoVo cells. In the alliein-treated group（3 and 6 μg/ml） , after 24 hours, the inhibition rates of migratory time were 24% and 50% （ t = 4. 543, 12. 348, P = 0. 010, 0. 001 ）, the inhibition rates of adhesion were 19% and 28% （ t = 6. 145, 6. 355, P - 0. 004, 0. 003 ） , the inhibition rates of migration were 28% and 46% （ t = 8.065, 28.435, both P 〈 0.01 ）, and the inhibition rates of invasion were 44% and 65% respectively （ t = 21. 274, 26. 288, both P 〈 0. 01 ）. Allicin at non-eytotoxic concentration eould down-regulate the mRNA levels of VEGF, uPAR and HPA in a dose-dependent manner in LoVo eells （t=7. 129, 6. 764, 8. 497, P = 0. 002, 0. 002, 0. 001 ） while the mRNA levels of TIMP-2, CD147 and nm23-H1 remained basically unchanged with the same treatment （t = 0. 341, 1. 889, 0. 914, P =0. 059, 0. 132, 0. 412）. The expression of MMP-2 had not been detected in LoVo cells. Conclusion Allicin in vitro inhibits invasion and metastasis of human colon carcinoma eell LoVo at non-cytotoxic concentration through down-regulating the expression of VEGF, uPAR and HPA mRNA.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第20期1382-1386,共5页
National Medical Journal of China
关键词
结肠肿瘤
大蒜
细胞增殖
细胞黏附
肿瘤转移
Colonic neoplasms
Garlic
Cell proliferation
Cell adhesion
Neoplasm metastasis
