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中华乳腺病杂志(电子版)

中华乳腺病杂志(电子版) ›› 2012, Vol. 06 ›› Issue (06) : 640 -648. doi: 10.3877/cma. j. issn.1674-0807.2012.06.006

论著

水通道蛋白3 在表皮生长因子诱导的乳腺癌细胞迁移中的作用机制研究
杨泉涌1, 张颖2, 张伟然1, 杨毅1, 牛瑞芳1, 张斌1,†(), 张霖1,†()   
  1. 1.300060 天津,天津医科大学附属肿瘤医院中心实验室 乳腺癌防治教育部重点实验室天津市肿瘤防治重点实验室
    2.300100 天津,天津市南开医院内科
  • 收稿日期:2012-06-27 出版日期:2012-12-01
  • 通信作者: 张斌, 张霖

Critical role of aquaporin-3 in epidermal growth factor-induced migration in breast cancer cells

Quan-yong YANG1, Ying ZHANG1, Wei-ran ZHANG1, Yi YANG1, Rui-fang NIU1, Bin ZHANG1,†(), Lin ZHANG1,†()   

  1. 1.Central Laboratory, Key Laboratory of Breast Cancer Prevention and Therapy of the Ministry of Education, Cancer Institute and Hospital, Tianjin Medical University, Tianjin 300060, China
  • Received:2012-06-27 Published:2012-12-01
  • Corresponding author: Bin ZHANG, Lin ZHANG
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杨泉涌, 张颖, 张伟然, 杨毅, 牛瑞芳, 张斌, 张霖. 水通道蛋白3 在表皮生长因子诱导的乳腺癌细胞迁移中的作用机制研究[J/OL]. 中华乳腺病杂志(电子版), 2012, 06(06): 640-648.

Quan-yong YANG, Ying ZHANG, Wei-ran ZHANG, Yi YANG, Rui-fang NIU, Bin ZHANG, Lin ZHANG. Critical role of aquaporin-3 in epidermal growth factor-induced migration in breast cancer cells[J/OL]. Chinese Journal of Breast Disease(Electronic Edition), 2012, 06(06): 640-648.

目的

研究水通道蛋白3(aquaporin 3,AQP3)与表皮生长因子(EGF)诱导的乳腺癌细胞迁移之间的关系,并对其可能的机制进行初步探索。

方法

采用Western blot 法检测4 种乳腺癌细胞MDA-MB-231、T47D、MCF-7、ER-ZR-70 中的AQP3与EGFR 的表达情况,选取高表达的细胞株进行EGF 诱导,观察细胞迁移(划痕愈合实验)及AQP3 表达的变化。 施加CuSO4(AQP3 抑制剂)后观察细胞迁移及AQP3 表达的变化。 为进一步证明EGF 通过哪条通路来调节AQP3 的表达,采用LY294002(PI3K/AKT 通路抑制剂)及U1026(MAPK/ERK1/2 通路抑制剂)分别进行阻断,观察AQP3 的表达变化。

结果

Western blot 检测发现迁移性强的MDA-MB-231 细胞具有EGFR 和AQP3 的高表达。

结论

在乳腺癌细胞中存在AQP3 表达;在EGF 诱导的乳腺癌细胞迁移中,AQP3 的反应性表达起到重要的作用,而EGF 可能通过PI3K/AKT 通路来调节AQP3 的表达。

关键词: 乳腺肿瘤, 水通道蛋白3, 表皮生长因子, 细胞迁移

Objective

To explore the relationship between aquaporin 3 (AQP3)expression and epidermal growth factor (EGF)induced cell migration in breast cancer cells.

Methods

AQP3 and EGF receptor(EGFR) expressions were tested by Western blot in four breast carcinoma cell lines MDA-MB-231, T47D, MCF-7 and ER-ZR-70. Cultured MDAMB-231 cells were treated with human EGF and subjected to cell migration assay (scratch test). The expression or activation level of proteins was analyzed by Western blot. CuSO4(AQP3 inhibitor) was applied to observe the cell migration and AQP3 expression. To further demonstrate the pathway by which EGF regulates AQP3 expression, LY294002 (PI3K/AKT pathway inhibitor) and U1026 (MAPK/ERK1/2 pathway inhibitor) were used separately to observe the changes in AQP3 expression.

Results

Western blot analysis found that MDAMB-231 cells, which had a strong potential of migration, showed the high expression of EGFR and AQP3. After EGF at different concentrations was used to induce the migration, it was found that AQP3 reactive expression was correlated with the degree of wound healing(r=0.885, P＜0.01). After CuSO4 at different concentrations was added to inhibit AQP3 expression, the degree of wound healing was decreased in MDA-MB -231 cells significantly(r=0.959, P＜0.01). The expression of AQP3 was reduced after LY294002 application,but presented no statistical difference after U1026 application.

Conclusions

AQP3 is expressed in breast cancer cells. AQP3 expression plays an important role in EGF-induced breast cancer cell migration and EGF could regulate AQP3 expression via PI3K/AKT pathway.

Key words: breast neoplasms, aquaporin 3, epidermal growth factor, cell migration
图1 Western blot 检测AQP3 在4 种乳腺癌细胞中的表达 1:MDA-MB-231;2:MCF-7;3:T47D;4:ER-ZR-70
图2 细胞划痕实验检测不同浓度EGF 诱导的MDA-MB-231 细胞迁移
图3 不同浓度EGF 诱导下的MDA-MB-231 细胞迁移率 a:P＜0.01,与EGF 0 ng/ml 相比,差异有统计学意义
图4 Western blot 检测EGF 诱导的MDA-MB-231 细胞中AQP3 的表达
图5 不同浓度EGF 诱导的MDA-MB-231 细胞中AQP3 的表达比率 a:P＜0.01,与EGF 0 ng/ml 组相比,差异有统计学意义
图6 细胞划痕实验检测不同浓度的CuSO4 抑制下EGF 40 ng/ml 组的MDA-MB-231 细胞迁移
图7 不同浓度的CuSO4 抑制下EGF 40 ng/ml 组的MDA-MB-231 细胞迁移率 a:P＜0.01,与对照组相比,差异有统计学意义
图8 Western blot 检测不同浓度的CuSO4 抑制下EGF 40 ng/ml组的MDA-MB-231 细胞中AQP3 的表达
图9 不同浓度的CuSO4 抑制下EGF 40 ng/ml 组的MDA-MB-231 细胞AQP3 的表达比率 a:P＜0.01,与对照组相比,差异有统计学意义
图10 Western blot 检测AQP3 表达的细胞通路 1:没有任何处理;2:只给予EGF 刺激;3:给予EGF 及LY294002 阻断;4:给予EGF 刺激及U1026 阻断;5:给予EGF 刺激及LY294002、U1026 阻断
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