Solasodine Induces Apoptosis, Affects Autophagy, and Attenuates Metastasis in Ovarian Cancer Cells

Abstract

Solasodine, a steroidal alkaloid isolated from solanaceous species, exhibits anticancer activities on several cell lines. This study aimed to explore the antitumor potential of solasodine on ovarian cancer cells. The MTT assay, lactate dehydrogenase release assay, Hoechst 33342 staining, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine staining assay, and Annexin V/PI assay were conducted to investigate the antiproliferation and apoptosis-inducing effects of solasodine. Monodansylcadaverine staining was performed to label the acidic puncta on ovarian cancer HEY cells. A wound healing assay and Transwell assay were carried out to determine whether solasodine elicits an antimetastatic effect on HEY cells. A gelatin zymography assay was applied to detect the enzymatic activities of matrix metalloproteinases. Western blot was employed to examine relevant protein expression. Results revealed that solasodine inhibited cell viabilities in a time- and dose-dependent manner, triggered apoptotic body formation, reduced cell mitochondrial membrane potential, and interfered with autolysosome degradation in ovarian cancer cells. Solasodine also suppressed the migration and invasion of HEY cells by downregulating matrix metalloproteinase expression and activities. This study could be used as a basis for further studies on the molecular mechanisms of the antiproliferation, apoptosis-inducing, autophagy-modifying, and antimetastatic activities of solasodine.

Key words

ovarian cancer - solasodine - apoptosis - autophagy - migration - invasion

• References

• 1 Allemani C, Weir HK, Carreira H, Harewood R, Spika D, Wang XS, Bannon F, Ahn JV, Johnson CJ, Bonaventure A, Marcos-Gragera R, Stiller C, Azevedo e Silva G, Chen WQ, Ogunbiyi OJ, Rachet B, Soeberg MJ, You H, Matsuda T, Bielska-Lasota M, Storm H, Tucker TC, Coleman MP. Group CW. Global surveillance of cancer survival 1995–2009: analysis of individual data for 25, 676, 887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet 2015; 385: 977-1010
• 2 Stewart BW, Wild CP. World Cancer Report, 2014. In: Stewart BW, Wild CP. eds. International Agency for Research of Cancer. France: International Agency for Research of Cancer; 2015: 678-697
• 3 Sayal K, Gounaris I, Basu B, Freeman S, Moyle P, Hosking K, Iddawela M, Jimenez-Linan M, Abraham J, Brenton J, Hatcher H, Earl H, Parkinson C. Epirubicin, cisplatin, and capecitabine for primary platinum-resistant or platinum-refractory epithelial ovarian cancer: results of a retrospective, single-institution study. Int J Gynecol Cancer 2015; 25: 977-984
• 4 Jayson GC, Kohn EC, Kitchener HC, Ledermann JA. Ovarian cancer. Lancet 2014; 384: 1376-1388
• 5 Gaikwad SM, Thakur B, Sakpal A, Singh RK, Ray P. Differential activation of NF-kappaB signaling is associated with platinum and taxane resistance in MyD88 deficient epithelial ovarian cancer cells. Int J Biochem Cell Biol 2015; 61: 90-102
• 6 Jin Y, Feng LP, Jiang X, Wang YX, Yin J, Yang ZP, Li Y, Pan LY. Annexin A3 is a potential predictor of platinum resistance in epithelial ovarian cancer patients in a prospective cohort. J Cancer 2015; 6: 678-685
• 7 Cui CZ, Wen XS, Cui M, Gao J, Sun B, Lou HX. Synthesis of solasodine glycoside derivatives and evaluation of their cytotoxic effects on human cancer cells. Drug Discov Ther 2012; 6: 9-17
• 8 Cham BE, Daunter B. Solasodine glycosides. Selective cytotoxicity for cancer cells and inhibition of cytotoxicity by rhamnose in mice with sarcoma 180. Cancer Lett 1990; 55: 221-225
• 9 Cham BE, Chase TR. Solasodine rhamnosyl glycosides cause apoptosis in cancer cells. Do they also prime the immune system resulting in long-term protection against cancer?. Planta Med 2012; 78: 349-353
• 10 Cham BE, Daunter B, Evans RA. Topical treatment of malignant and premalignant skin lesions by very low concentrations of a standard mixture (BEC) of solasodine glycosides. Cancer Lett 1991; 59: 183-192
• 11 Punjabi S, Cook LJ, Kersey P, Marks R, Cerio R. Solasodine glycoalkaloids: a novel topical therapy for basal cell carcinoma. A double-blind, randomized, placebo-controlled, parallel group, multicenter study. Int J Dermatol 2008; 47: 78-82
• 12 Lee KR, Kozukue N, Han JS, Park JH, Chang EY, Baek EJ, Chang JS, Friedman M. Glycoalkaloids and metabolites inhibit the growth of human colon (HT29) and liver (HepG2) cancer cells. J Agric Food Chem 2004; 52: 2832-2839
• 13 Sharma T, Airao V, Panara N, Vaishnav D, Ranpariya V, Sheth N, Parmar S. Solasodine protects rat brain against ischemia/reperfusion injury through its antioxidant activity. Eur J Pharmacol 2014; 725: 40-46
• 14 Abdel-Latif L, Murray BK, Renberg RL, OʼNeill KL, Porter H, Jensen JB, Johnson FB. Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis. J Gen Virol 2006; 87: 2539-2548
• 15 Petit PX, Lecoeur H, Zorn E, Dauguet C, Mignotte B, Gougeon ML. Alterations in mitochondrial structure and function are early events of dexamethasone-induced thymocyte apoptosis. J Cell Biol 1995; 130: 157-167
• 16 Cossarizza A, Baccarani-Contri M, Kalashnikova G, Franceschi C. A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1). Biochem Biophys Res Commun 1993; 197: 40-45
• 17 Chen Z, Zhang Y, Yang J, Jin M, Wang XW, Shen ZQ, Qiu Z, Zhao G, Wang J, Li JW. Estrogen promotes benzo[a]pyrene-induced lung carcinogenesis through oxidative stress damage and cytochrome c-mediated caspase-3 activation pathways in female mice. Cancer Lett 2011; 308: 14-22
• 18 Biederbick A, Kern HF, Elsasser HP. Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuoles. Eur J Cell Biol 1995; 66: 3-14
• 19 Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, Overvatn A, Bjorkoy G, Johansen T. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 2007; 282: 24131-24145
• 20 Matsubara M, Zieske JD, Fini ME. Mechanism of basement membrane dissolution preceding corneal ulceration. Invest Ophthalmol Vis Sci 1991; 32: 3221-3237
• 21 Cham BC. Solasodine rhamnosyl glycosides specifically bind cancer cell receptors and induce apoptosis and necrosis treatment for skin cancer and hope for internal cancers. Res J Biol Sci 2007; 2: 503-514
• 22 Cham BC. Drug therapy: Solamargine and other solasodine rhamnosyl glycosides as anticancer agents. Modern Chemother 2013; 2: 33-49
• 23 Mizushima N, Komatsu M. Autophagy: renovation of cells and tissues. Cell 2011; 147: 728-741
• 24 Rebecca VW, Amaravadi RK. Emerging strategies to effectively target autophagy in cancer. Oncogene 2016; 35: 1-11
• 25 Xi G, Hu X, Wu B, Jiang H, Young CY, Pang Y, Yuan H. Autophagy inhibition promotes paclitaxel-induced apoptosis in cancer cells. Cancer Lett 2011; 307: 141-148
• 26 Zhang R, Wang R, Chen Q, Chang H. Inhibition of autophagy using 3-methyladenine increases cisplatin-induced apoptosis by increasing endoplasmic reticulum stress in U251 human glioma cells. Mol Med Rep 2015; 12: 1727-1732
• 27 Lamberti M, Porto S, Zappavigna S, Stiuso P, Tirino V, Desiderio V, Mele L, Caraglia M. Levofolene modulates apoptosis induced by 5-fluorouracil through autophagy inhibition: clinical and occupational implications. Int J Oncol 2015; 46: 1893-1900
• 28 Tang MC, Wu MY, Hwang MH, Chang YT, Huang HJ, Lin AM, Yang JC. Chloroquine enhances gefitinib cytotoxicity in gefitinib-resistant nonsmall cell lung cancer cells. PLoS One 2015; 10: e0119135
• 29 Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 2000; 19: 5720-5728
• 30 Johansen T, Lamark T. Selective autophagy mediated by autophagic adapter proteins. Autophagy 2011; 7: 279-296
• 31 Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, Overvatn A, Stenmark H, Johansen T. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 2005; 171: 603-614
• 32 Overall CM, Kleifeld O. Tumour microenvironment – opinion: validating matrix metalloproteinases as drug targets and anti-targets for cancer therapy. Nat Rev Cancer 2006; 6: 227-239
• 33 Kamat AA, Fletcher M, Gruman LM, Mueller P, Lopez A, Landen jr. CN, Han L, Gershenson DM, Sood AK. The clinical relevance of stromal matrix metalloproteinase expression in ovarian cancer. Clin Cancer Res 2006; 12: 1707-1714
• 34 Wang Y, Ye Y, Lin J, Meyer L, Wu X, Lu K, Liang D. Genetic variants in matrix metalloproteinase genes as disposition factors for ovarian cancer risk, survival, and clinical outcome. Mol Carcinog 2015; 54: 430-439
• 35 Lock R, Kenific CM, Leidal AM, Salas E, Debnath J. Autophagy-dependent production of secreted factors facilitates oncogenic RAS-driven invasion. Cancer Discov 2014; 4: 466-479
• 36 Zhan Z, Xie X, Cao H, Zhou X, Zhang XD, Fan H, Liu Z. Autophagy facilitates TLR4- and TLR3-triggered migration and invasion of lung cancer cells through the promotion of TRAF6 ubiquitination. Autophagy 2014; 10: 257-268
• 37 Galavotti S, Bartesaghi S, Faccenda D, Shaked-Rabi M, Sanzone S, McEvoy A, Dinsdale D, Condorelli F, Brandner S, Campanella M, Grose R, Jones C, Salomoni P. The autophagy-associated factors DRAM1 and p62 regulate cell migration and invasion in glioblastoma stem cells. Oncogene 2013; 32: 699-712
• 38 Catalano M, DʼAlessandro G, Lepore F, Corazzari M, Caldarola S, Valacca C, Faienza F, Esposito V, Limatola C, Cecconi F, Di Bartolomeo S. Autophagy induction impairs migration and invasion by reversing EMT in glioblastoma cells. Mol Oncol 2015; 9: 1612-1625
• 39 Xu J, Yang H, Zhou X, Wang H, Gong L, Tang C. Bisdemethoxycurcumin suppresses migration and invasion of highly metastatic 95D lung cancer cells by regulating E-cadherin and vimentin expression, and inducing autophagy. Mol Med Rep 2015; 12: 7603-7608
• 40 Wu GS, Song YL, Yin ZQ, Guo JJ, Wang SP, Zhao WW, Chen XP, Zhang QW, Lu JJ, Wang YT. Ganoderiol A-enriched extract suppresses migration and adhesion of MDA-MB-231 cells by inhibiting FAK-SRC-paxillin cascade pathway. PLoS One 2013; 8: e76620