In Vitro and In Vivo Antitumor Activities of Tenacissoside C from Marsdenia tenacissima

Abstract

Tenacissoside C, a natural bioactive compound of C₂₁ steroidal saponins, was isolated and purified from air-dried stems of Marsdenia tenacissima. In the present study, the MTT assay showed that tenacissoside C exhibited obvious cytotoxicity in K562 cells with IC₅₀ values of 31.4, 22.2, and 15.1 µM for 24, 48, and 72 h, respectively. Flow cytometry analysis indicated that the antiproliferative activity induced by tenacissoside C might be executed via G0/G1 cell cycle arrest and proapoptosis in K562 cells. Western blotting analysis elucidated that: A) Tenacissoside C induced K562 cell cycle (G0/G1) arrest via downregulating cycline D1 protein expression; and B) Tenacissoside C induced K562 cell apoptosis via the mitochondrial pathway by downregulating Bcl-2 and Bcl-xL protein expression, upregulating Bax and Bak protein expression, and activating caspase-9 and caspase-3. In vivo, significant tumor growth inhibition activity of tenacissoside C was observed in K562 cell-bearing nude mice, accompanied by a significant antiangiogenic effect in vivo against K562 cells (a marked decrease in MVD) and associated with enhanced apoptotic cell death (TUNEL staining assay in vivo), both in dose-dependent manners. The treatment with tenacissoside C did not significantly affect body mass and macroscopic examination of the organs in this mouse tumor model.

Key words

Marsdenia tenacissima - Asclepiadaceae - tenacissoside C - antitumor - cytotoxicity - K562 cell - apoptosis

• References

• 1 Banerji V, Frumm SM, Ross KN, Li LS, Schinzel AC, Hahn CK, Kakoza RM, Chow KT, Ross L, Alexe G, Tolliday N, Inguilizian H, Galinsky I, Stone RM, DeAngelo DJ, Roti G, Aster JC, Hahn WC, Kung AL, Stegmaier K. The intersection of genetic and chemical genomic screens identifies GSK-3alpha as a target in human acute myeloid leukemia. J Clin Invest 2012; 122: 935-947
• 2 Santos FP, Bueso-Ramos CE, Ravandi F. Acute erythroleukemia: diagnosis and management. Expert Rev Hematol 2010; 3: 705-718
• 3 Cailleteau C, Micallef L, Lepage C, Cardot PJ, Beneytout JL, Liagre B, Battu S. Investigating the relationship between cell cycle stage and diosgenin-induced megakaryocytic differentiation of HEL cells using sedimentation field-flow fractionation. Anal Bioanal Chem 2010; 398: 1273-1283
• 4 Leger DY, Liagre B, Beneytout JL. Role of MAPKs and NF-kappaB in diosgenin-induced megakaryocytic differentiation and subsequent apoptosis in HEL cells. Int J Oncol 2006; 28: 201-207
• 5 Cragg GM, Newman DJ. Nature: a vital source of leads for anticancer drug development. Phytochem Rev 2009; 8: 313-331
• 6 Ruan WJ, Lai MD, Zhou JG. Anticancer effects of Chinese herbal medicine, science or myth?. J Zhejiang Univ Sci B 2006; 7: 1006-1014
• 7 Cao QQ, Zhang RS. The study of chemical composition and pharmacological function of Marsdenia plants. Chin J Mod Drug Appl 2008; 2: 1-5
• 8 Li HY, Wang W, Dong FY. Advances in studies on chemical constituents in stem of Marsdenia tenacissima and their pharmacological effects. Chin Tradit Herb Drugs 2007; 38: 1101-1104
• 9 Zhang MZ, He Z, Wu GY. Effect of xiaoaiping injection on Ec-9706 cell line. Lishizhen Med Mater Med Res 2008; 19: 1182-1184
• 10 Wang SM, Zhao HP. Inhibitory effect of xiaoaiping injection combined with octreotide acetate on transplanted liver cancer in mice. Chin J Clin Oncol 2011; 38: 1135-1138
• 11 Yu SS, Chen MC, Li ZX, Ji YB, Wu B. Progress on chemical and pharmacological studies of Marsdenia tenacissima . Chin J Exp Tradit Med Formul 2011; 17: 279-283
• 12 Shobha S, Maheshwari PK, Anakshi K. Cissogenin, a pregnane genin from Marsdenia tenacissima . Phytochemistry 1980; 19: 2427-2430
• 13 Shuji M, Kimiko Y, Koji H, Koh K, Hiroshi M. Five glycosides from Chinese drug ʼTong-Guang-Sanʼ: the stems of Marsdenia tenacissima . Phytochemistry 1986; 25: 2861-2865
• 14 Xia ZH, Xing WX, Mao SL, Lao AN, Uzawa J, Yoshida S. Pregnane glycosides from the stems of Marsdenia tenacissima . J Asian Nat Prod Res 2004; 6: 79-85
• 15 Deng J, Liao ZX, Chen DF. Marsdenosides A–H, polyoxypregnane glycosides from Marsdenia tenacissima . Phytochemistry 2005; 66: 1040-1051
• 16 Wang XL, Li QF, Yu KB, Peng SL, Zhou Y, Ding LS. Four new pregnane glycosides from the stems of Marsdenia tenacissima . Helv Chim Acta 2006; 89: 2738-2744
• 17 Zhang H, Tan AM, Zhang AY, Chen R, Yang SB, Huang X. Five new C21 steroidal glycosides from the stems of Marsdenia tenacissima . Steroids 2010; 75: 176-183
• 18 Huang XD, Liu T, Wang S. Two new polyoxypregnane glycosides from Marsdenia tenacissima . Helv Chim Acta 2009; 92: 2111-2117
• 19 Koumtebaye E, Su N, Hu WF, Lin SS, Sun L, Yuan ST. Antitumor activity of Xiaoaiping injection on human gastric cancer SGC-7901 cells. Chin J Nat Med 2012; 10: 339-346
• 20 Hu Y J, Shen XL, Lu HL, Zhang YH, Huang XA, Fu LC, Fong WF. Tenacigenin B derivatives reverse p-glycoprotein-mediated multidrug resistance in HepG2/Dox cells. Nat Prod 2008; 71: 1049-1051
• 21 Sun B, Geng S, Huang X, Zhu J, Liu S, Zhang Y, Ye J, Li Y, Wang J. Coleusin factor exerts cytotoxic activity by inducing G0/G1 cell cycle arrest and apoptosis in human gastric cancer BGC-823 cells. Cancer Lett 2011; 301: 95-105
• 22 Sreelatha S, Jeyachitra A, Padma PR. Antiproliferation and induction of apoptosis by Moringa oleifera leaf extract on human cancer cells. Food Chem Toxicol 2011; 49: 1270-1275
• 23 Hsiao WL, Liu L. The role of traditional Chinese herbal medicines in cancer therapy–from TCM theory to mechanistic insights. Planta Med 2010; 76: 1118-1131
• 24 Fulda S. Modulation of apoptosis by natural products for cancer therapy. Planta Med 2010; 76: 1075-1079
• 25 Yoon JH, Gores GJ. Death receptor-mediated apoptosis and the liver. J Hepatol 2002; 37: 400-410
• 26 Sharma PR, Mondhe DM, Muthiah S, Pal HC, Shahi AK, Saxena AK, Qazi GN. Anticancer activity of an essential oil from Cymbopogon flexuosus . Chembiol Interact 2009; 179: 160-168
• 27 Mignotte B, Vayssiere JL. Mitochondria and apoptosis. Eur J Biochem 1998; 252: 1-15
• 28 Tepper CG, Seldin MF, Mudryj M. Fas-mediated apoptosis of proliferating, transiently growth-arrested, and senescent normal human fibroblasts. Exp Cell Res 2000; 260: 9-19
• 29 Zhang MC, Liu HP, Demchik LL, Zhai YF, Yang DJ. Light sensitizes IFN-gamma-mediated apoptosis of HT-29 human carcinoma cells through both death receptor and mitochondria pathways. Cell Res 2004; 14: 117-124
• 30 Hsu HF, Houng JY, Kuo CF, Tsao N, Wu YC. Glossogin, a novel phenylpropanoid from Glossogyne tenuifolia, induced apoptosis in A549 lung cancer cells. Food Chem Toxicol 2008; 46: 3785-3791
• 31 Liu ZB, Hou YF, Di GH, Wu J, Shen ZZ, Shao ZM. PA-MSHA inhibits proliferation and induces apoptosis through the up-regulation and activation of caspases in the human breast cancer cell lines. J Cell Biochem 2009; 108: 195-206
• 32 Reed JC. Regulation of apoptosis by Bcl-2 family proteins and its role in cancer and chemoresistance. Curr Opin Oncol 1995; 7: 541-546
• 33 Bruce-Keller AJ, Begley JG, Fu W, Butterfield DA, Bredesen DE, Hutchins JB, Hensley K, Mattson MP. Bcl-2 protects isolated plasma and mitochondrial membranes against lipid peroxidation induced by hydrogen peroxide and amyloid beta-peptide. J Neurochem 1998; 70: 31-39
• 34 Kim CF, Jiang JJ, Leung KN, Fung KP, Lau CB. Inhibitory effects of Agaricus blazei extracts on human myeloid leukemia cells. J Ethnopharmacol 2009; 122: 320-326
• 35 Sun CM, Huang SF, Zeng JM, Liu DB, Xiao Q, Tian WJ, Zhu XD, Huang ZG, Feng WL. Per2 inhibits K562 leukemia cell growth in vitro and in vivo through cell cycle arrest and apoptosis induction. Pathol Oncol Res 2010; 16: 403-411
• 36 Weidner N. Intratumor microvessel density as a prognostic factor in cancer. Am J Pathol 1995; 147: 9-19