目的

探讨阿司匹林对HER-2 阳性乳腺癌AU-565 细胞增殖的影响。

方法

将AU-565细胞分为2 组,其中一组细胞用阿司匹林处理,对照组用DMSO 处理。 用MTT 法检测不同浓度阿司匹林溶液(0.05、0.01、0.25、0.50、1.00、2.50、5.00、10.00 mmol/L)对AU-565 细胞增殖能力的影响;用平板克隆形成实验检测2.50 mmol/L 阿司匹林溶液处理2 周后对AU-565 细胞克隆形成能力的影响;用Western blot 检测5.00 mmol/L 阿司匹林溶液处理AU-565 细胞72 h 后细胞增殖及凋亡指标的表达变化。 MTT 实验、平板克隆形成实验及Western blot 结果数据比较采用t 检验。

结果

MTT 实验显示,与对照组相比,5.00 mmol/L 及10.00 mmol/L 阿司匹林组的细胞增殖率均明显受到抑制[(82.9±6.5)%比(59.5±13.1)%, t=3.580, P=0.007;(86.9±14.5)% 比(17.8±3.2)%, t=10.443, P＜0.001]。 平板克隆形成实验显示,阿司匹林组克隆形成率仅有(0.5±0.1)%,低于对照组的克隆形成率(87.9±4.7)%(t=33.406,P＜0.050)。 Western blot 实验结果显示:阿司匹林组中增殖指标磷酸化腺苷酸活化蛋白激酶α、磷酸化mTOR、c-myc 的相对表达量分别为2.20±0.38、0.57±0.12、0.55±0.05,与对照组(0.98±0.22、1.13±0.32、1.32±0.31)相比,差异均具有统计学意义(t=4.890、2.803、4.285,P 均＜0.050);阿司匹林组中凋亡指标剪切的聚腺苷二磷酸核糖聚合酶和髓样细胞白血病-1 的相对表达量(1.10±0.14、1.85±0.12)均高于对照组(0.62±0.12、0.92±0.25)(t=4.539、5.680,P 均＜0.050)。

结论

阿司匹林可抑制HER-2 阳性乳腺癌AU-565 细胞的增殖能力。

Objective

To investigate the effect of aspirin on the proliferation of human HER-2 positive breast cancer AU-565 cells.

Methods

The AU-565 cells were cultured and divided into two groups, the experimental group was given aspirin at different concentrations and the control group was given DMSO as control. In MTT assay, AU-565 cells were treated with different concentrations of aspirin (0.05,0.01,0.25,0.50,1.00,2.50,5.00,10.00 mmol/L) to measure the proliferation of cells. Using colony formation assay,the colony formation ability of AU-565 cells was determined 2 weeks after treatment of 2.50 mmol/L aspirin.Cells were harvested 72 h after aspirin (5.00 mmol/L) treatment to examine the expressions of cell proliferation and apoptosis indexes with Western blot method. The results in MTT assay, colony formation assay and Western blot experiment were compared by t test.

Results

MTT assay showed that compared with control group, the proliferation rate was significantly inhibited in aspirin-treated group at 5.00 mmol/L [(82.9±6.5)% vs(59.5±13.1)%, t=3.580, P=0.007] and 10.00 mmol/L [86.9±14.5)% vs(17.8±3.2)%,t=10.443, P＜0.001]. The colony formation rate of AU-565 cells was (0.5±0.1) % in aspirin-treated group, significantly lower than (87.9±4.7)% in control group (t=33.406, P＜0.050). The relative protein levels of proliferation indexes (p-AMPKα,p-mTOR and c-myc) were 2.20±0.38,0.57±0.12,0.55±0.05 in aspirin-treated group, 0.98±0.22, 1.13±0.32, 1.32±0.31 in control group, suggesting a significant difference(t = 4.890, 2.803, 4.285; all P ＜0.050). The relative protein levels of apoptosis indexes(cleaved-PARP and Mcl-1) were 1.10±0.14, 1.85±0.12 in aspirin-treated group, significantly higher than 0.62±0.12,0.92±0.25 in control group respectively (t=4.539,5.680;both P＜0.050).

Conclusion

Aspirin inhibits the proliferation ability of HER-2 positive breast cancer AU-565 cells.