Polysaccharide from Sepia esculenta ink and cisplatin inhibit synergistically proliferation and metastasis of triple-negative breast cancer MDA-MB-231 cells

Document Type: Original Article


1 College of Chemistry and Environment, Guangdong Ocean University, Zhanjiang 524088, China

2 Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning 530001, China

3 College of Food Science & Technology, Guangdong Ocean University, Zhanjiang 524088, China

4 Institute of Food Science & Engineering Technology, Hezhou University, Hezhou 542899, China

5 Experimental Center for Sciences, Guilin Medical University, Guilin 541004, China

6 Biochemisty Center, Guangdong Ocean University, Zhanjiang 524088, China


Objective(s): This paper aims to investigate synergistic inhibition of polysaccharide from Sepia esculenta ink (SIP), a newly isolated marine polysaccharide in our laboratory, on breast cancer MDA-MB-231 cells exposed to cisplatin.
Materials and Methods: Cell viability of MDA-MB-231 cells was determined by CCK 8 assay. Median-effect concentration was analyzed using Chou-Talalay method that was also subjected to determine cell inhibition ratio and combined index, as well as interaction between SIP and cisplatin. Proliferation and migration abilities were detected with plate colony formation assay and cell wound scratch assay, respectively. Expression of MMP-2 and MMP-9 proteins was measured with Western blot assay.
Results: Data showed that SIP not only suppressed proliferation and migration of MDA-MB-231 cells, and expression of MMP-2 and MMP-9 proteins, also promoted inhibition of cisplatin on proliferation, migration and MMPs expression of MDA-MB-231 cells, which indicates synergy inhibition of drug combination of SIP and cisplatin on breast cancer cells. The median-effect concentrations of cisplatin and SIP were 4.9 and 1659.6 μg/ml, respectively. Whereas the concentration of combination drug was 158.5 μg/ml. The data indicated that drug combination can decrease dosages of the two single agents, especially the usual dosage of cisplatin.
Conclusion: This research demonstrated that SIP repressed proliferation and metastasis of MDA-MB-231 cells and promoted anticancer effect of cisplatin on the breast cancer cells. The data suggested that SIP is a potential natural drug that can be used as an auxiliary medicine alongside chemotherapy in treating breast cancer.


1.Smith L, Welham KJ, Watson MB, Drew PJ, Lind MJ, Cawkwell L. The protemic analysis of cisplatin resistance in breast cancer cells. Oncol Res 2007; 16: 497-506.

2.Martin M. Platinum compounds in the treatment of advanced breast cancer. Clin Breast Cancer 2001; 2: 190-208.

3.Vassilomanolakis M, Koumakis G, Barbounis V, Demiri M, Panopoulos C, Chrissohoou M, et al. First-line chemotherapy with docetaxel and cisplatin in metastatic breast cancer. Breast 2005; 4:136-141.

4.Garand C, Guay D, Sereduk C, Chow D, Tsofack SP, Langlois M, et al. An integrative approach to identify YB-1-interacting proteins required for cisplatin resistance in MCF-7 and MDA-MB-231 breast cancer cells. Cancer Sci 2011; 102: 1410-1417.

5.Luo P, Liu HZ. Antioxidant ability of squid ink polysaccharides as well as their protective effects on DNA damage in vitro. Afr J Pharm Pharmacocl 2013; 7: 1382-1388.

6.Le XY, Luo P, Gu YP, Tao YX, Liu HZ. Squid ink polysaccharide reduces cyclophosphamide-induced testicular damage via Nrf2/ARE activation pathway in mice. Iran J Basic Med Sci 2015; 18: 827-831.

7.Le XY, Luo P, Gu YP, Tao YX, Liu HZ. Interventional effects of squid ink polysaccharide on cyclophos-phamide-associated testicular damage in mice. Bratisl Med J 2015; 116: 334-339.

8.Liu C, Li X, Li Y, Feng Y, Zhou S, Wang F. Structural characterization and antimutagenic activity of a novel polysaccharide isolated from Sepiella maindroni ink. Food Chem 2008; 110: 807-813.

9.Chen S, Wang J, Xue C, Li H, Sun B, Xue Y, et al. Sulfation of a squid ink polysaccharide and its inhibitory effect on tumor cell metastasis. Carbohyd Polym 2010; 81:560-566.

10.Zong A, Liu Y, Zhang Y, Song X, Shi Y, Cao H, et al. Anti-tumor activity and the mechanism of SIP-S: A sulfated polysaccharide with anti-metastatic effect. Carbohyd Polym 2015; 129: 50-54.

11.Zong A, Zhao T, Zhang Y, Song X, Shi Y, Cao H, et al. Anti-metastatic and anti-angiogenic activities of sulfated polysaccharide of Sepiella Maindroni ink. Carbohyd Polym 2013; 91:403-409.

12.Zuo T, Cao L, Xue C, Tang QJ. Dietary squid ink polysaccharide induces goblet cells to protect small intestine from chemotherapy induced injury. Food Funct 2015; 6:981-986.

13.Zuo T, He X, Cao L, Xue C, Tang QJ. The dietary polysaccharide from Ommastrephes bartrami prevents chemotherapeutic mucositis by promoting the gene expression of antimicrobial peptides in Paneth cells. J Funct Food 2015; 12:530-539.

14.Zuo T, Cao L, Li X, Zhang Q, Xue C, Tang Q. The squid ink polysaccharide protect tight junctions and adherens junctions from chemotherapeutic injury in the small intestinal epithelium of mice. Nutr Cancer 2015; 67: 364-371.

15.Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 2006; 58: 621-681.

16.Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay Method. Cancer Res 2010; 70:440-446.

17.Liu HZ, Tao YX, Luo P, Deng CM, Gu YP, Yang L, et al. Preventive effects of a novel polysaccharide from Sepia esculenta ink on ovarian failure and its action mechanisms in cyclophosphamide-treated mice. J Agr Food Chem 2016; 64: 5759-5766.

18.Dawood S. Triple-negative breast cancer: Epidemiology and management options. Drugs 2010; 70:2247-2258.

19.Ressler S, Mlineritsch B, Greil R. Triple negative breast cancer. Memo 2010; 3:185-189.

20.Hatzopoulos S, Di Stefano M, Campbell KM, Falgione D, Ricci D, Rosignoli M, et al. Cisplatin ototoxicity in the Sprague Dawley rat evaluated by distortion product otoacoustic emissions. Audiology 2001; 40:253-264.

21.Liedert B, Materna V, Schadendorf D, Thomale J, Lage H. Overexpression of cMOAT (MRP2/ABCC2) is associated with decreased formation of platinum-DNA adducts and decreased G2-arrest in melanoma cells resistant to cisplatin. J Invest Dermatol 2003; 121:172-176.

22.Jia Y, Zhang C, Zhou L, Xu H, Shi Y, Tong Z. Micheliolide overcomes KLF 4-mediated cisplatin resistance in breast cancer cells by downregulating glutathione. OncoTargets Ther 2015; 8:2319-2327.

23.Radin D, Lippa A, Patel P, Leonardi D. Lifeguard inhibition of Fas-mediated apoptosis: a possible mechanism for explaining the cisplatin resistance of triple-negative breast cancer cells. Biomed Pharmacother 2016; 77:161-166.

24.Takaya Y, Uchisawa H, Narumi F, Matsue H. Illexins A, B, and C from squid ink should have a branched structure. Biochem Biophys Res Commun 1996; 226:335-338.

25.Chen SG, Xu J, Xue CH, Dong P, Sheng WJ, Yu GL, et al. Sequence determination of a non-sulfated glycosaminoglycan-like polysaccharide from melanin-free ink of the squid Ommastrephes bartrami by negative-ion electrospray tandem mass spectrometry and NMR spectroscopy. Glycoconjugate J 2008; 25:481-492.

26.Thorns V, Walter GF, Thorns C. Expression of MMP-2, MMP-7, MMP-9, MMP-10 and MMP-11 in human astrocytic and oligodendroglial gliomas. Anticancer Res 2003; 23:3937-3944.