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Journal of ZheJiang University(Medical Science)  2013, Vol. 42 Issue (5): 538-542    DOI: 10.3785/j.issn.1008-9292.2013.05.010
    
PP2 enhances intercellular communication of gap junction in breast cancer Hs578T cells
DONG Shu-Ying, ZHENG Chao, JIANG Guo-Jun, HAN Xi, TONG Xu-Hui
Department of Pharmacology,Faculty of Pharmacy,Bengbu Medical College,Bengbu 233000,China
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Abstract  

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/β-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: PP2 enhances the gap junction function in breast cancer Hs578T cells,which is probably related to the inhibition of Src kinase.



Key wordsBreast neoplasms      src-Family kinases/antagonists &      inhibitors      Connexins      Tumor cells, Cultured     
Received: 17 December 2012      Published: 25 September 2013
CLC:  R 74  
Cite this article:

DONG Shu-Ying, ZHENG Chao, JIANG Guo-Jun, HAN Xi, TONG Xu-Hui. PP2 enhances intercellular communication of gap junction in breast cancer Hs578T cells. Journal of ZheJiang University(Medical Science), 2013, 42(5): 538-542.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2013.05.010     OR     http://www.zjujournals.com/med/Y2013/V42/I5/538


PP2可增强乳腺癌Hs578T细胞缝隙连接功能

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;Western blot法检测PP2对乳腺癌细胞Src激酶表达的影响。
结果:1~8 μmol/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,1~8 μmol/L的PP2能显著增强细胞荧光传递功能(P<0.01),8 μmol/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),8 μmol/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。


关键词: 乳腺肿瘤,  src族激酶类/拮抗剂和抑制剂,  连接蛋白类,  肿瘤细胞, 培养的 
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