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Inhibition of Gastric Cancer Cells Invasion and Metastasis by Zileuton in Viro

Author: ShiJianHua
Tutor: LiuChunLun
School: Chongqing Medical University
Course: Internal Medicine
Keywords: SGC-7901 cells The cyclooxygenase -2/5- lipoxygenase Attack Shift
CLC: R735.2
Type: Master's thesis
Year: 2010
Downloads: 30
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Abstract


Objective To observe the cyclooxygenase -2/5- lipoxygenase (cycloxygenase-2/5- 1ipoxygenase, COX-2/5-LOX) dual inhibitor Darbufelone on gastric cancer SGC-7901 cell growth, heterogeneous adhesion, invasion induced angiogenesis, migration ability. Initial explore possible mechanisms Darbufelone affect SGC-7901 cell invasion and metastasis. Method 1.MTT assay SGC-7901 cell growth and adhesion ability to change. 2.Transwell Chamber membrane invasion and migration assay and invasion of SGC-7901 cells and vertical migration capabilities change. 3. The scratches experimental observation SGC-7901 cells lateral migration change. 4. Observed SGC-7901 cells induced by conditioned medium of human umbilical vein endothelial cells ECV304 formation changes in the ability of the lumen. 5.RT-PCR detection of osteopontin in SGC-7901 cells (Osteopontin, OPN) and matrix metalloproteinase -2 (Matrix metalloproteinases-2, MMP-2) mRNA expression changes. 6. Immunocytochemistry and Western blotting SGC-7901 cells in OPN and MMP-2 protein expression changes. 1. 1.5 x 10-5 mol / L of Darbufelone 72 h treatment, SGC-7901 cell growth was significantly decreased (P lt; 0.05). 2. 1.5 x 10-5 mol / L of Darbufelone after 72 h of treatment, 30,60,90,120 min SGC-7901 cell adhesion rate were (27.10 ± 3.44)%, (46.36 ± 4.52)%, (50.37 ± 7.90)% and (58.01 ± 9.22)%, control group (34.99 ± 2.12)%, (60.60 ± 4.89)%, (70.65 ± 5.20)%, (84.62 ± 7.52)% in the treatment group at each time point Compared with the control group showed a significant difference (P lt; 0.05), inhibition rates were (22.20 ± 5.58)%, (23.57 ± 1.61)%, (28.98 ± 5.97)%, (31.79 ± 4.52)%. 3.Transwell Chamber invasion and migration experiments showed that: after 1.5 x 10-5 mol / L of Darbufelone for 72 h after the invasion of SGC-7901 cells penetrating cells (50.00 ± 3.67) and migration penetrating cells (93.00 ± 6.56), compared with the control group (82.00 ± 7.14, 143.00 ± 9.82) was significantly reduced (P lt; 0.01). 4 scratch test: 1.5 x 10-5 mol / L of Darbufelone after 72 h of treatment, SGC-7901 cells 24 h, 48 h migration distance was shorter than that in the control group. Angiogenesis experiments showed that: 1.5 × 10-5 mol / L Darbufelone for 72 h subsequent formation of SGC-7901 induced human umbilical vein endothelial cell line ECV304 the lumen number of lumen length was 82.70 ± 4.58 ( 172.20 ± 6.41) μm, the control group was 142.00 ± 5.86 (214.60 ± 9.44) μm, the treatment group compared with control group significant difference (P lt; 0.01). 6.RT-PCR: 1.5 x 10-5 mol / L Darbufelone role in SGC-7901 after 72 h of OPN and MMP-2 mRNA (0.34 ± 0.03,0.33 ± 0.04) than in the control group (0.54 ± 0.01, 0.57 ± 0.09) was significantly lower (P lt; 0.05). 7. OPN and MMP-2 protein expression: 1) immunocytochemistry Show: After 1.5 × 10-5 mol / L Darbufelone for 72 h, SGC-7901 OPN and MMP-2 protein AOD values ??(0.18 ± 0.01,0.26 ± 0.02) compared with the control group (0.26 ± 0.03,0.33 ± 0.03) was significantly lower (P lt; 0.05). 2) Western blotting: after 1.5 × 10-5 mol / L Darbufelone role in SGC-7901 after 72 h OPN protein expression (0.27 ± 0.02) compared with the control group (0.44 ± 0.06) was significantly lower (P lt; 0.01); MMP-2 protein expression (0.35 ± 0.03) compared to the control group (0.48 ± 0.06) was significantly lower (P lt; 0.05). Conclusion COX-2/5-LOX dual inhibitor the Darbufelone can effectively inhibit the growth of SGC-7901 cells, invasion, migration and the ability to induce neovascularization. Darbufelone inhibition of SGC-7901 cell invasion and metastasis may be related to the inhibition of OPN and MMP-2 expression.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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