Effect of pycnogenol on proliferation, migration and metastasis of human breast cancer MCF-7 cells

doi:10.3877/cma.j.issn.1674-0807.2019.03.006

Abstract

Abstract:

Objective

To investigate the inhibitory effect of the antioxidant pycnogenol on human breast cancer MCF-7 cells, and to observe the effect of pycnogenol on the metastasis and migration of MCF-7 cells.

Methods

MCF-7 cells were treated with 10, 20, 40 μg/ml pycnogenol, respectively, as experimental groups. The control group was only treated with blank DMEM medium (without fetal bovine serum). The optical density of MCF-7 cells at the wavelength of 490 nm was determined by the MTT method at 24, 48 and 72 h after treatment. The apoptosis rate of MCF-7 cells was measured by the flow cytometry. The senescence rate of MCF-7 cells was measured by β-galactosidase staining. The effect of pycnogenol at different concentrations on the migration and invasion of MCF-7 cells were detected by Transwell chamber assay. The expression of senescence-associated proteins P53, P21, P16, P27 and E2F1 in MCF-7 cells was determined by Western blot analysis. The expression of matrix metalloproteinase (MMP)-2 and MMP-9 was also determined by Western blot analysis. The number of migrated cells and metastatic cells, and the protein expression among four groups were compared by one-factor analysis of variance. The optical density was compared by repeated measurement analysis of variance and pairwise comparison was performed by LSD method.

Results

The result of MTT assay showed that at 24, 48, and 72 h after treatment, the optical density of breast cancer MCF-7 cells was significantly different among four groups (F=149.439, P<0.001) and at different time points (F=27.922, P<0.001); there was an interaction between grouping and time points (F=18.466, P<0.001). The result of flow cytometry showed that the apoptosis rate of MCF-7 cells was (2.36±0.27)%, (6.44±1.43)%, (7.52±2.09)% and (11.68±1.65)% in the blank control group, 10, 20 and 40 μg/ml pycnogenol group, respectively, indicating a significant difference (F=19.143, P<0.001). The result of β-galactosidase staining showed that the senescence rate of MCF-7 cells was (5.35±1.32)%, (20.08±2.14)%, (40.55±4.61)% and (59.26±4.10)% in the blank control group, 10, 20 and 40 μg/ml pycnogenol group, respectively, indicating a significant difference (F=150.150, P<0.001). The result of Transwell chamber assay showed that the number of migrated cells was 41±5, 27±2, 19±1 and 11±2 in the blank control group, 10, 20 and 40 μg/ml pycnogenol group, respectively, indicating a significant difference (F=59.330, P<0.001); the number of metastatic cells was 24±4, 17±1, 12±2 and 7±2, respectively in the blank control group, 10, 20 and 40 μg/ml pycnogenol group, respectively, indicating a significant difference (F=26.230, P<0.001). Western blot results showed that pycnogenol up-regulated the expression of P53, P21, P16 and P27 (F=263.905, 424.937, 217.515, 391.115, all P<0.001), and down-regulated the expression of E2F1 (F=64.003, P<0.001); the expression levels of MMP-2 and MMP-9 in MCF-7 cells were significantly decreased (F=44.104, 45.594, both P<0.001).

Conclusion

Pycnogenol may inhibit the growth of breast cancer MCF-7 cells by promoting the cellular senescence and it can also inhibit the cell migration and metastasis.

Key words: Breast neoplasms, Cellular senescence, Migration, Metastasis, Pycnogenol

Chunshu Yang, Guangyuan Qin, Xinyu Zheng. Effect of pycnogenol on proliferation, migration and metastasis of human breast cancer MCF-7 cells[J]. Chinese Journal of Breast Disease(Electronic Edition), 2019, 13(03): 165-172.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhrxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0807.2019.03.006

https://zhrxbzz.cma-cmc.com.cn/EN/Y2019/V13/I03/165

Figures/Tables 9

References 36

[1]	薛静，王浩. 乳腺癌免疫治疗的研究进展[J/CD]. 中华乳腺病杂志(电子版)，2018，12 (1):43-49.
[2]	Huynh HT, Teel RW. Selective induction of apoptosis in human mammary cancer cells (MCF-7) by pycnogenol [J]. Anticancer Res, 2000, 20(4): 2417-2420.
[3]	Ruocco N, Costantini S, Guariniello S, et al. Polysaccharides from the marine environment with pharmacological, cosmeceutical and nutraceutical potential[J]. Molecules, 2016, 21(5): E551.
[4]	梁璟慧，吴毓东，杨丽萍．三阴性乳腺癌新辅助化疗的最新进展[J/CD]．中华乳腺癌病杂志(电子版)，2016, 9(4):270-274.
[5]	马哿，王佳，夏添松，等. 淋巴细胞趋化因子对乳腺癌MCF-7细胞的表柔比星药物敏感性及侵袭转移能力的影响[J/CD]. 中华乳腺病杂志(电子版)，2017, 11(6):354-360.
[6]	Collado M, Serrano M. Senescence in tumours: evidence from mice and humans[J]. Nat Rev Cancer, 2010, 10(1): 51-57.
[7]	Nardella C, Clohessy JG, Alimonti A, et al. Pro-senescence therapy for cancer treatment[J]. Nat Rev Cancer, 2011, 11(7): 503-511.
[8]	Campisi J. Replicative senescence: an old lives’tale? [J]. Cell, 1996, 84(4): 497-500.
[9]	Campisi J, d’Adda di Fagagna F. Cellular senescence: when bad things happen to good cells[J]. Nat Rev Mol Cell Biol, 2007, 8(9): 729-740.
[10]	Campisi J. Aging, cellular senescence, and cancer[J]. Annu Rev Physiol, 2013, 75: 685-705.
[11]	Collado M, Blasco MA, Serrano M. Cellular senescence in cancer and aging[J]. Cell, 2007, 130(2): 223-233.
[12]	刘翠翠，王光学，赵倩，等. 长链非编码RNA调控乳腺癌增殖、转移、耐药性及乳腺癌干细胞[J/CD]. 中华乳腺病杂志(电子版)，2016, 10(5):310-315.
[13]	Rodier F, Campisi J. Four faces of cellular senescence[J]. J Cell Biol, 2011, 192(4): 547-556.
[14]	Coppé JP, Desprez PY, Krtolica A, et al. The senescence-associated secretory phenotype: the dark side of tumor suppression[J]. Annu Rev Pathol, 2010, 5: 99-118.
[15]	Coppé JP, Patil CK, Rodier F, et al. Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor[J]. PLoS Biol, 2008, 6(12): 2853-2868.
[16]	Rodier F, Coppé JP, Patil CK, et al. Persistent DNA damage signaling triggers senescence-associated inflammatory cytokine secretion[J]. Nat Cell Biol, 2009, 11(8): 973-979.
[17]	George BP, Abrahamse H. A review on novel breast cancer therapies: photodynamic therapy and plant derived agent induced cell death mechanisms[J]. Anticancer Agents Med Chem, 2016, 16(7):793-801.
[18]	Newman DJ, Cragg GM. Natural products as sources of new drugs over the 30 years from 1981 to 2010[J]. J Nat Prod，2012, 75 (3): 311-335.
[19]	Kaczirek K, Schindl M, Weinhäusel A, et al. Cytotoxic activity of camptothecin and paclitaxel in newly established continuous human medullary thyroid carcinoma cell lines[J]. J Clin Endocrinol Metab, 2004, 89(5):2397-2401.
[20]	Popolo A, Pinto A, Daglia M, et al. Two likely targets for the anti-cancer effect of indole derivatives from cruciferous vegetables: PI3K/Akt/mTOR signalling pathway and the aryl hydrocarbon receptor[J]. Semin Cancer Biol，2017，46:132-137.
[21]	Aiyer HS, Warri AM, Woode DR, et al. Influence of berry polyphenols on receptor signaling and cell-death pathways: implications for breast cancer prevention[J]. J Agri Food Chem, 2002, 60(23):5693-5708.
[22]	Zhang X, Chen LX, Ouyang L, et al. Plant natural compounds: targeting pathways of autophagy as anti-cancer therapeutic agents[J]. Cell Prolif, 2002, 45(5): 466-476.
[23]	Reuben SC, Gopalan A, Petit DM, et al. Modulation of angiogenesis by dietary phytoconstituents in the prevention and intervention of breast cancer[J]. Mol Nutr Food Res, 2012, 56(1): 14-29.
[24]	Horakova L, Licht A, Sandig G, et al. Standardized extracts of flavonoids increase the viability of PC12 cells treated with hydrogen peroxide: effects on oxidative injury[J]. Arch Toxicol, 2003，77(1):22-29.
[25]	Peng QL, Buz’Zard AR, Lau BH. Pycnogenol protects neurons from amyloid-beta peptide-induced apoptosis[J]. Brain Res Mol Brain Res, 2002，104(1):55-65.
[26]	Roseff SJ. Improvement in sperm quality and function with French maritime pine tree bark extract[J]. J Reprod Med, 2002, 47(10):821-824.
[27]	Rohdewald P. A review of the French maritime pine bark extract (pycnogenol), a herbal medication with a diverse clinical pharmacology[J]. Int J Clin Pharm Ther, 2002, 40(4):158-168.
[28]	Nelson AB, Lau BH, Ide N, et al. Pycnogenol inhibits macrophage oxidative burst, lipoprotein oxidation and hydroxyl radical-induced DNA damage[J]. Drug Dev Indust Pharm, 1998, 24(2): 139-144.
[29]	Wei ZH, Peng QL, Lau BH. Pycnogenol enhances endothelial cell antioxidant defenses[J]. Redox Rep，1997，3(4): 219-224.
[30]	Huang WW, Yang JS, Lin CF, et al. Pycnogenol induces differentiation and apoptosis in human promyeloid leukemia HL-60 cells[J]. Leuk Res, 2005, 29(6): 685-692.
[31]	Buz’Zard AR, Lau BH. Pycnogenol reduces talcinduced neoplastic transformation in human ovarian cell cultures[J]. Phytother Res, 2007, 21(6): 579-586.
[32]	Belcaro G, Cesarone MR, Genovesi D, et al. Pycnogenol may alleviate adverse effects in oncologic treatment[J]. Panminerva Med，2008，50(3):227-234.
[33]	Egeblad M, Werb Z. New functions for the matrix metalloproteinases in cancer progression[J]. Nat Rev Cancer, 2002, 2(3):161-174.
[34]	Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment[J]. Cell, 2010, 141(1): 52-67.
[35]	Yoon SO, Park SJ, Yun CH, et al. Roles of matrix metalloproteinases in tumor metastasis and angiogenesis[J]. J Biochem Mol Biol, 2003, 36(1):128-137.
[36]	Kim D, Rhee S. Matrix metalloproteinase-2 regulates MDA-MB-231 breast cancer cell invasion induced by active mammalian diaphanous-related formin 1[J]. Mol Med Rep, 2016, 14(1): 277-282.

组别	24 h	48 h	72 h
空白对照组	0.918±0.028	1.005±0.043	1.122±0.142
10 μg/ml碧萝芷组	0.887±0.038	0.770±0.015^a	0.650±0.013^a
20 μg/ml碧萝芷组	0.712±0.031^ab	0.652±0.020^ab	0.510±0.030^ab
40 μg/ml碧萝芷组	0.721±0.052^ab	0.556±0.032^abc	0.334±0.033^abc

组别	24 h	48 h	72 h
空白对照组	0.918±0.028	1.005±0.043	1.122±0.142
10 μg/ml碧萝芷组	0.887±0.038	0.770±0.015^a	0.650±0.013^a
20 μg/ml碧萝芷组	0.712±0.031^ab	0.652±0.020^ab	0.510±0.030^ab
40 μg/ml碧萝芷组	0.721±0.052^ab	0.556±0.032^abc	0.334±0.033^abc

组别	实验次数	细胞数目(个，±s)
组别	实验次数	迁移	侵袭
空白对照组	3	41±5	24±4
10 μg/ml碧萝芷组	3	27±2^a	17±1^a
20 μg/ml碧萝芷组	3	19±1^ab	12±2^ab
40 μg/ml碧萝芷组	3	11±2^abc	7±2^abc
F值	?	59.330	26.230
P值	?	<0.001	<0.001

组别	实验次数	细胞数目(个，±s)
组别	实验次数	迁移	侵袭
空白对照组	3	41±5	24±4
10 μg/ml碧萝芷组	3	27±2^a	17±1^a
20 μg/ml碧萝芷组	3	19±1^ab	12±2^ab
40 μg/ml碧萝芷组	3	11±2^abc	7±2^abc
F值	?	59.330	26.230
P值	?	<0.001	<0.001

组别	实验次数	P53	P21	P16	P27	E2F1
空白对照组	3	1.000±0.000	1.000±0.000	1.000±0.000	1.000±0.000	1.000±0.000
10 μg/ml碧萝芷组	3	1.252±0.045^a	1.309±0.045^a	1.244±0.024^a	1.309±0.036^a	0.821±0.051^a
20 μg/ml碧萝芷组	3	1.618±0.036^ab	1.796±0.059^ab	1.422±0.048^ab	1.825±0.042^ab	0.688±0.033^ab
40 μg/ml碧萝芷组	3	1.941±0.067^abc	2.021±0.022^abc	1.782±0.056^abc	1.954±0.055^abc	0.532±0.061^abc
F值	?	263.905	424.937	217.515	391.115	64.003
P值	?	<0.001	<0.001	<0.001	<0.001	<0.001

Please choose a citation manager

Content to export