Effect of RORα Overexpression on Proliferation, Migration and Invasion of Human Gastric MGC803 Cells
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摘要:目的
观察RORα高表达对MGC803细胞增殖与迁移侵袭的影响。
方法构建高表达RORα人胃癌MGC803细胞。Real-time PCR或RT-PCR和Western blot检测RORα、MMP-9与TIMP3 mRNA和蛋白表达。MTT、流式细胞术、迁移和侵袭实验检测RORα高表达对MGC803细胞增殖、细胞周期、迁移和侵袭能力的影响。
结果RORα高表达MGC803细胞RORα mRNA和蛋白表达显著上调。在48、72与96 h，RORα高表达细胞的增殖活性明显低于对照组和空载体组（P＜0.05）。RORα高表达G2/M期细胞明显高于对照组和空载体组（P＜0.05）。高表达组细胞的迁移距离较对照组与空载体组明显减少（P＜0.05）。高表达组穿膜细胞数较对照组与空载体组明显减少（P＜0.05）。RORα高表达可明显下调MMP-9和上调TIMP3 mRNA与蛋白。
结论成功构建RORα高表达MGC803细胞，RORα高表达可抑制MGC803细胞增殖与迁移侵袭，将细胞阻滞于G2/M期，其机制可能与下调MMP-9和上调TIMP3有关。
Abstract:ObjectiveTo investigate the effect of RORα overexpression on human gastric cancer MGC803 cells proliferation,migration and invasion.
MethodsThe expressions of RORα,MMP-9 and TIMP3 mRNA and protein were detected by Real-time PCR or RT-PCR and Western blot. The effect of RORα overexpression on the proliferation,cell cycle,migration and invasion of MGC803 cells were detected by MTT,flow cytometry,wound healing and Transwell assays.
ResultsThe expressions of RORα mRNA and protein were stably increased in the RORα/MGC803 cells. The proliferation activity in RORα/MGC803 cells were notably lower than in MGC803 cells and in vector/MGC803,respectively,at 48 h,72 h and 96 h(P＜0.05) . Percentage of G2/M in RORα/MGC803 cells markedly higher than that in MGC803 cells and vector/MGC803(P＜0.05) . The migration length in RORα/MGC803 cells was markedly lower than that in MGC803 and vector/MGC803 cells (P＜0.05) . The cells through the matrigel coated membrane in RORα/MGC803 significantly decreased comparing with MGC803 and vector/MGC803 cells(P＜0.05) . RORα overexpression was downregulation of MMP-9 and upregulation of TIMP3 mRNA and proteins(P＜0.05) . RORα overexpression could significantly downregulate MMP-9 and upregulate TIMP3 mRNA and proteins(P＜0.05) .
ConclusionRORα/MGC803 cells with stably overexpressed RORα are successfully constructed. The overexpression of RORα may inhibit the proliferation,migration and invasion,as well as G2/M arrest in MGC803 cells,which may be correlated with the downregulation of MMP-9 and the upregulation of TIMP3.
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Key words:
- Human gastric MGC803 cells /
- RORα /
- Eukaryotic expression vector /
- Proliferation /
- Migration /
- Invasion
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图 3 RORα高表达MGC803细胞中RORα蛋白的表达
Figure 3 Expression of RORα protein in RORα/MGC803 cells
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图 4 RORα高表达对MGC803细胞的细胞周期影响
Figure 4 Effect of RORα overexpression on MGC803 cells cycle
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图 5 RORα高表达对MGC803细胞迁移的影响
Figure 5 Effect of RORα overexpression on migration of MGC803 cells
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图 6 RORα高表达对胃癌MGC803细胞侵袭的影响
Figure 6 Effect of RORα overexpression on invasion of MGC803 cells
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图 7 RORα高表达对MGC803细胞MMP-9 mRNA表达的影响
Figure 7 Effect of RORα overexpression on MMP-9 mRNA expression in MGC803 cells
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图 8 RORα高表达对MGC803细胞MMP-9蛋白表达的影响
Figure 8 Effect of RORα overexpression on MMP-9 protein expression in MGC803 cells
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图 9 RORα高表达对MGC803细胞TIMP3 mRNA表达的影响
Figure 9 Effect of RORα overexpression on TIMP3 mRNA expression in MGC803 cells
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图 10 RORα高表达对MGC803细胞TIMP3蛋白表达的影响
Figure 10 Effect of RORα overexpression on TIMP3 protein expression in MGC803 cells
表 1 Real-time PCR检测RORα高表达MGC803细胞中RORα mRNA表达
Table 1 Expression of RORα mRNA in RORα/MGC803 cells detected by Real-time PCR
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表 2 RORα高表达对MGC803细胞增殖的影响
Table 2 Effect of RORα overexpression on proliferation of MGC803 cells
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