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作 者:董淑英[1] 郑超[1] 蒋国君[1] 韩溪[1] 童旭辉[1]
机构地区:[1]蚌埠医学院药学系药理学教研室,安徽蚌埠233030
出 处:《浙江大学学报(医学版)》2013年第5期538-542,582,共6页Journal of Zhejiang University(Medical Sciences)
基 金:国家自然科学基金(81001457);安徽省教育厅省级优秀青年人才基金重点项目(2010SQRL121ZD);安徽省教育厅高校省级科学研究项目(kj2010B117)
摘 要:目的:探讨Src激酶抑制剂PP2对乳腺癌细胞Hs578T缝隙连接功能的影响.方法:MTT法检测PP2对乳腺癌细胞生长的影响;细胞接种荧光示踪法观察PP2对乳腺癌细胞之间荧光传递功能的影响;Westem blot法检测PP2对乳腺癌细胞Sre激酶表达的影响.结果:1~8μmol/L浓度的PP2对乳腺癌细胞生长几乎无影响;细胞接种荧光示踪结果显示,1~8μmol/L的PP2能显著增强细胞荧光传递功能(P<0.01),8μmol/L的PP2作用6、12和24 h后细胞荧光传递功能亦明显增强(P< 0.01);Western blot结果显示,PP2(1~8 μmoL/L)可显著降低Src激酶表达(P<0.05 ~0.01),8μmol/L PP2作用6、12和24 h后Src激酶的表达亦明显降低(P<0.01).结论:PP2能增强乳腺癌细胞Hs578T的缝隙连接功能,这种增强作用可能与其降低Src激酶表达有关.Objective:To investigate the effect of Src kinase inhibitor PP2 on intercellular communication of gap junction in breast cancer cells.Methods:Cultured breast cancer Hs578T cells were treated with various concentrations of pp2 (0,1,2,4,8,16,32 μmol/L) for 24h.Cell growth was determined by MTT assay; dye spread in Hs578T cells was measured by Parachute assay; and the expression of Src kinase in Hs578T cells was detected by Western blot.Results:MTT assay showed that the survive rate of Hs578T cells treated with PP2 (1 ~8 μmoL/L) was 98% ± 3% ~ 94% ± 4%.Parachute assay showed that compared to control group the standard normalized dye spread rates of Hs578T cells treated with 1,2,4 and 8 μmol/L PP2 were 1.60 ±0.08,2.00 ±0.05,2.20 ±0.05 and 2.70±0.09,respectively (all P <0.01).Moreover,compared to control group at the same time points,the standard normalized dye spread of Hs578T cells treated with 8 μmol/L PP2 for 6,12 and 24 h were 1.4 ± 0.05,1.7 ± 0.06,and 2.2 ± 0.07,respectively (all P < 0.01).Western blot showed that the expression ratios of Src kinase/β-actin of Hs578T cells treated with 1,2,4 and 8 μmol/L PP2 for 24 h were 0.93 ± 0.02,0.70 ± 0.09,0.66 ± 0.09 and 0.36 ± 0.10,which were significantly inhibited compared with control group (P < 0.05 or 0.01).And the expression ratio of Src kinase/β3-actin of Hs578T cells treated with 8 μmol/L PP2 for 6,12 and 24h was 0.82 ±0.03,0.66 ±0.08 and 0.59 ±0.09,which were all inhibited significantly compared to control group (P < 0.01).Conclusion:PP2enhances the gap junction function in breast cancer Hs578T cells,which is probably related to the inhibition of Src kinase.
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