Influence of aspirin on proliferation of HER-2 positive breast cancer AU-565 cells

doi:10.3877/cma.j.issn.1674-0807.2017.01.004

Abstract

Abstract:

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.

Key words: Breast neoplasms, Receptor, erbB-2, Aspirin, Cell proliferation, Apoptosis

Ying Wu, Zhe Wang, Jing Kong, Jian Zhang, Rui Ling. Influence of aspirin on proliferation of HER-2 positive breast cancer AU-565 cells[J]. Chinese Journal of Breast Disease(Electronic Edition), 2017, 11(01): 19-23.

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References 25

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组别	试验次数	0.05mmol/L	0.10mmol/L	0.25mmol/L	0.50mmol/L	1.00mmol/L	2.50mmol/L	5.00mmol/L	10.00mmol/L
对照组	3	91.3±12.0	109.8±21.9	83.6±9.8	72.3±15.2	68.5±4.0	70.7±8.8	82.9±6.5	86.9±14.5
阿司匹林组	3	103.3±13.7	114.2±12.5	93.7±29.2	72.8±19.6	74.6±9.3	61.7±6.1	59.5±13.1	17.8±3.2
t值		1.472	0.395	0.524	0.051	1.332	1.899	3.580	10.443
P值		0.179	0.703	0.490	0.963	0.218	0.094	0.007	＜0.001

组别	试验次数	0.05mmol/L	0.10mmol/L	0.25mmol/L	0.50mmol/L	1.00mmol/L	2.50mmol/L	5.00mmol/L	10.00mmol/L
对照组	3	91.3±12.0	109.8±21.9	83.6±9.8	72.3±15.2	68.5±4.0	70.7±8.8	82.9±6.5	86.9±14.5
阿司匹林组	3	103.3±13.7	114.2±12.5	93.7±29.2	72.8±19.6	74.6±9.3	61.7±6.1	59.5±13.1	17.8±3.2
t值		1.472	0.395	0.524	0.051	1.332	1.899	3.580	10.443
P值		0.179	0.703	0.490	0.963	0.218	0.094	0.007	＜0.001

组别	实验次数	ERK	p-ERK	c-myc	AKT
对照组	3	1.17±0.27	0.79±0.18	1.32±0.31	0.27±0.06
阿司匹林组	3	1.45±0.35	1.37±0.33	0.55±0.05	0.57±0.12
t值		1.123	2.685	4.285	4.016
P值		0.324	0.055	0.013	0.016
组别	实验次数	p-AKT	AMPKα	p-AMPKα	m-TOR
对照组	3	0.84±0.22	1.49±0.26	0.98±0.22	1.10±0.23
阿司匹林组	3	1.19±0.30	1.77±0.34	2.20±0.38	1.30±0.27
t值		1.644	1.114	4.890	0.963
P值		0.175	0.328	0.008	0.390
组别	实验次数	p-mTOR	Mcl-1	cleaved PARP	PARP
对照组	3	1.13±0.32	0.92±0.25	0.62±0.12	0.97±0.19
阿司匹林组	3	0.57±0.12	1.85±0.12	1.10±0.14	1.30±0.23
t值		2.803	5.680	4.539	1.936
P值		0.049	0.005	0.011	0.125

组别	实验次数	ERK	p-ERK	c-myc	AKT
对照组	3	1.17±0.27	0.79±0.18	1.32±0.31	0.27±0.06
阿司匹林组	3	1.45±0.35	1.37±0.33	0.55±0.05	0.57±0.12
t值		1.123	2.685	4.285	4.016
P值		0.324	0.055	0.013	0.016
组别	实验次数	p-AKT	AMPKα	p-AMPKα	m-TOR
对照组	3	0.84±0.22	1.49±0.26	0.98±0.22	1.10±0.23
阿司匹林组	3	1.19±0.30	1.77±0.34	2.20±0.38	1.30±0.27
t值		1.644	1.114	4.890	0.963
P值		0.175	0.328	0.008	0.390
组别	实验次数	p-mTOR	Mcl-1	cleaved PARP	PARP
对照组	3	1.13±0.32	0.92±0.25	0.62±0.12	0.97±0.19
阿司匹林组	3	0.57±0.12	1.85±0.12	1.10±0.14	1.30±0.23
t值		2.803	5.680	4.539	1.936
P值		0.049	0.005	0.011	0.125

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