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[摘要]
目的:探讨丹参酮IIA 磺酸钠(sodium tanshinone IIA sulfonate,STS)对宫颈癌HeLa 细胞迁移的影响及其机制。方法:Transwell 迁移实验检测不同浓度STS、整合素β3 特异性抗体LM609 和转染过表达整合素β3 质粒对宫颈癌HeLa细胞迁移的影响,Western blotting 法检测不同浓度STS 和STS 刺激不同时间对HeLa 细胞整合素β3 表达的影响,CCK-8 法检测STS 对HeLa细胞活力的影响。结果:STS 以浓度依赖的方式降低HeLa 细胞的迁移(P<0.01),LM609 显著降低HeLa 细胞迁移率(P<0.01),STS刺激转染过表达整合素β3 质粒HeLa细胞的迁移率较单一STS刺激和STS刺激转染空质粒HeLa细胞的迁移率显著增加(P<0.01);STS以浓度和时间依赖的方式降低HeLa细胞整合素β3 的表达(P<0.05);不同浓度STS刺激的HeLa细胞存活数和空白对照组相比无显著差异(P>0.05)。结论:STS可减少宫颈癌HeLa细胞整合素β3 的表达,STS可显著减少HeLa细胞的迁移,整合素β3 特异性抗体LM609 也可显著降低HeLa细胞的迁移,转染过表达整合素β3 质粒则可减少STS对HeLa细胞迁移的抑制,STS可能通过减少整合素β3 的表达抑制HeLa细胞迁移,这为进一步阐明STS抗肿瘤的分子机制和STS临床应用提供实验依据。
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[Abstract]
Objective: To explore effect of sodium tanshinone IIA sulfonate (STS) on migration of the cervix carcinoma Hela cell and its possible mechanism. Methods:Effects of STS in different concentrations, specific antibody to integrin β3 (LM609), and transfection with integrin β3 over-expression plasmid on migration of the cervix carcinoma HeLa cell were detected by Transwell assay. Immunoblotting test was used to detect effects of STS in different concentrations and different times of STS stimulation on expression of integrin β3 in the HeLa cell. Effect of STS on viability of the HeLa cell was tested by CCK-8 assay. Results:STS in concentration-dependent manner reduced significantly migration of the HeLa cell (P<0.01). LM609 remarkably decreased migration rate of the HeLa cell (P<0.01). Migration rate of the HeLa cell transfected with over-expression integrin β3 plasmid and stimulated by STS was markedly more increase than those of the HeLa cell stimulated by STS only as well as the HeLa cell transfected with empty plasmid and stimulated by STS (P<0.01). STS in concentration- and timing-dependent manners reduced expression of integrin β3 in the HeLa cell (P<0.05). There was no significant difference in number of the viable cells between the HeLa cell stimulated with different concentrations of STS groups and the HeLa cells in blank control group (P>0.05). Conclusion:STS could reduce expression of integrin β3 in the cervix carcinoma HeLa cell, and significantly decrease migration of the HeLa cell,LM609 which is a specific antibody against integrin β3 could also reduced migration of the HeLa cells, and transfection with integrin β3 over- expression plasmid could reduce the inhibition of STS on migration of the HeLa cell. The results of this paper shown that STS might inhibits migration of the HeLa cell via decrease of integrin β3 expression, which might provide theoretical basis and experimental evidence for further explain molecular mechanism of STS anti-carcinoma and clinical application of STS.
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