Research Article | Volume: 8, Issue: 11, November, 2018

Cytotoxic effects and anti-proliferative cancer activity of coelomic fluid from Lumbricus rubellus promotes apoptosis and reduces G2/M phase progression in HT-29 cells

Sofy Permana Pearlindah Zurniatus Sholihah Agustin Iskandar Hani Susanti Agustina Tri Endharti   

Open Access   

Published:  Nov 30, 2018

DOI: 10.7324/JAPS.2018.81104

This study has investigated the effect of chemotherapy drugs combined Coelomic Fluid (CF) derived from Lumbricus rubellus in HT-29 cells. HT-29 cells were treated with a combination of 5-fluorouracil (5-FU) (5 μg/ml) and Coelomic Fluid Lumbricus (CFL) (5 μg/ml, 10 μg/ml, or 20 μg/ml), respectively. The cells were incubated for 24 hours and then analyzed by flow cytometry to assess apoptosis, cell cycle progression, and proliferation. The cytotoxicity effect was measured by MTT assay that causes 50% decrease in cell viability (IC50 value). Results showed that the percentage of apoptosis in combined-therapy groups significantly higher than 5-FU single therapy group (p < 0.001). Additionally, the percentage of proliferation and cell growth significantly inhibited of 5-FU and CFL combination groups (p < 0.001). However, cell cycle slightly reduced the percentage of G2/M phase. Our results showed that CFL inhibited cell proliferation that is strictly dependent on colon cancer. These findings suggest that CFL have cytotoxicity effect to inhibit cell growth, anti-proliferative activity and increased apoptosis against human colon cancer cells.

Keyword:     Apoptosis cell cycle cell proliferation cytotoxic effect coelomic fluid 5-FU.


Permana S, Pearlindah, Sholihah Z, Iskandar A, Susanti H, Endharti AT. Cytotoxic effects and anti-proliferative cancer activity of coelomic fluid from Lumbricus rubellus promotes apoptosis and reduces G2/M phase progression in HT-29 cells. J App Pharm Sci, 2018; 8(11): 028-034.

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

HTML Full Text


Niederhuber JE, Armitage JO, Doroshow JH, Tepper JE. 2014. Abeloff's Clinical Oncology Fifth edition. Philadelphia, PA 19103-2899.

Au-Yeung KK, Ko JK. Novel herbal flavonoids promote apoptosis but differentially induce cell cycle arrest in human colon cancer cell. Investigational New Drugs, 2010; 28(1):1-13.

Chalfant CE, Rathman K, Pinkerman RL, Wood RE, Obeid LM, Ogretmen B, Hannun YA. De novo ceramide regulates the alternative splicing of caspase 9 and Bcl-x in A549 lung adenocarcinoma cells. Dependence on protein phosphatase-1. J Biol Chem, 2002; 277:12587- 12595.

Chen H, Takahashi S, Imamura M, Okutani E, Zhang Z, Chayama K, Chen BA. Earthworm fibrinolytic enzyme: anti-tumor activity on human hepatoma cells in vitro and in vivo. Chin Med J, 2007; 120:898-904.

Chua W, Kho PS, Moore MM, Charles KA, Clarke SJ. Clinical, laboratory and molecular factors predicting chemotherapy efficacy and toxicity in colorectal cancer. Crit Rev Oncol Hematol, 2011; 79(3):224-250.

Ciccolini J, Gross E, Dahan L, Lacarelle B, Mercier C. Routine dihydropyrimidine dehydrogenase testing for anticipating 5-fluorouracil-related severe toxicities hype or hope? Clin Colorectal Cancer, 2010; 9(4):24-8.

Cooper EL, Balamurugan M, Huang CY, Tsao CR, Heredia J, Tommaseo-Ponzetta M, Paoletti MG. Earthworms dilong: Ancient, inexpensive, noncontroversial models my help clarify approaches to integrated medicine emphasizing neuroimmuno systems. Evid Based Complement Alternat Med, 2012; 2012:1-11.

De Colibus L, Sonnen AFP, Morris KJ, Siebert CA, Abrusci P, Plitzko J, Hodnik V, Leippe M, Volpi E, Anderluh G, Gilbert RJ. Structures of Lysenin Reveal a Shared Evolutionary Origin for Pore-Forming Proteins and its Mode of Sphingomyelin Recognition. Structure, 2012; 20(9):1498- 507.

Dinesh MS, Sridhar S, Chandana PG, Pai V, Geetha KS, Hegdge RN. Anticancer Potentials of Peptides of Coelomic Fluid of Earthworm Eudrilus eugeniae. Biotech Res Asia, 2013; 10(2):601-606.

Endharti AT, Adisti W, Anik L, Eviana N, Sofy P. Dendrophthoe pentandra (L.) Miq Extract Effectively Inhibits Inflammation, Proliferation and Induces p53 Expression. BMC, 2016; 16(374):1-8.

Endharti AT, Baskoro AD, Norahmawati E, Therapeutic effect of soluble worm protein acting as immune regulatory on colitis. Asian Pac J Trop Biomed, 2017a; 7(1):70-77.

Endharti AT and Permana. S. Extract from mango mistletoes Dendrophthoe pentandra ameliorates TNBS induced colitis by regulating CD4+ T cells in mesenteric lymph nodes. BMC Complementary and Alternative Medicine, 2017b; 17:468.

Endharti AT, Permana S. T-Bet Is Dependent on Stat-4 Inhibiting Acute Colitis but Not Stat-1 Using L4 Somatic Antigen of Heligmosomoides polygyrus. The Scientific World Journal, 2018; 2018:1-10.

Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer, 2010; 127(12):2893-2917.

Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F. Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer, 2015; 136(5):359-386.

Ghosh N, Patel N, Jiang K, Watson JE, Cheng J, Chalfant CE, Cooper DR. Ceramide-activated protein phosphatase involvement in insulin resistance via Akt, serine/arginine-rich protein 40, and ribonucleic acid splicing in L6 skeletal muscle cells. Endocrinology, 2007; 148:1359-1366.

Horssen RV, Timo LM, Alexander MM. TNF-α in Cancer Treatment: Molecular Insights, Antitumor Effects and Clinical Utility. The Oncologist, 2006; 11:394-408.

Jaabir MMS, Shamsheerali L, Yasar MMD, Kumar SS. Evaluation of the cell-free coelomic fluid of the earthworm Eudrilus euginiae to induce apoptosis in SiHa cell line. Journal of Pharmacy Research, 2011; 4(10):3417-3420.

Kobayashi H, Ohta N, Umeda M. Biology of Lysenin, a Protein in the Coelomic Fluid of the Earthworm Eisenia foetida. Japan. International Review of Cytology, 2004; 236:45-99.

Kracikova M, Akiri G, George A, Sachidanandam R, Aaronson SA. A threshold mechanism mediates p53 cell fate decision between growth arrest and apoptosis. Cell Death Differ, 2013, 20:576-588.

Kraveka JM, Li L, Bielawski J, Obeid LM, Ogretmen B. Involvement of endogenous ceramide in the inhibition of telomerase activity and induction of morphologic differentiation in response to alltrans-retinoic acid in human neuroblastoma cells. Arch Biochem Biophys, 2003; 419:110-119.

Mancikova V. 2011. Molecular Characterization of Selected Defense Factors in Lumbricidae. Prague; Charles University. Thesis.

Medema RH, Macůrek L. Checkpoint recovery in cells: how a molecular understanding can help in the fight against cancer. F1000 Biology Reports, 2011; 3:10.

Mishra J, Dromund J, Quazi SH, Karanki SS, Shaw JJ, Chen B, Kumar N. Prospective of Colon Cancer Treatments and Scope for Combinatorial Approach to Enhanced Cancer Cell Apoptosis. Crit Rev Oncol Hematol, 2013; 86(3):232-250.

Ogretmen B, Schady D, Usta J, Wood R, Kraveka JM, Luberto C, Birbes H, Hannun YA, Obeid LM. Role of ceramide in mediating the inhibition of telomerase activity in A549 human lung adenocarcinoma cells. J Biol Chem, 2001; 276:24901-24910.

Qiu T, Xiang X, Lei W, Hao C, Zhang L, Feng M, Yu F, Li J, Xiong J. Establishment and biological characterization of 5-fluorouracil-resistant human colon cancer HT-29/5-FU cell line. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2015; 31(3):328-32.

Redondo-Blanco S, Fernández J, Gutiérrez-del-Río I, Villar CJ, Lombó F. New Insights toward Colorectal Cancer Chemotherapy Using Natural Bioactive Compounds. Front Pharmacol, 2017; 8: 109.

Vidya N, Dinesh MS, Ananda S, Radha DK. Cytotoxic Potential of Eudrilus eugeniae coelomcyte culture supernatant against tumor cells. International Journal of Scientific and Research Publications, 2016; 6(8):2250-3153.

Watanabe T, Itabashi M, Shimada Y, Tanaka S, Ito Y, Ajioka Y, Hamaguchi T, Hyodo I, Igarashi M, Ishida H, Ishihara S, Ishiguro M, Kanemitsu Y, Kokudo N, Muro K, Ochiai A, Oguchi M, Ohkura Y, Saito Y, Sakai Y, Ueno H, Yoshino T, Boku N, Fujimori T, Koinuma N, Morita T, Nishimura G, Sakata Y, Takahashi K, Tsuruta O, Yamaguchi T, Yoshida M, Yamaguchi N, Kotake K, Sugihara K; Japanese Society for Cancer of the Colon and Rectum. Japanese Society for Cancer of the Colon and Rectum (JSCCR) Guidelines 2014 for treatment of colorectal cancer. Int J Clin Oncol, 2015; 20:207-239.

Zhang H, Yan-Hong W, Hong-Wei C, Li-Jun P, Yu-Dong C. Purification of a protein from coelomic fluid of the earthworm Eisenia foetida and evaluation of its hemolytic, antibacterial, and antitumor activities. Pharmaceutical Biology, 2011; 49(3):269-275. 034

Article Metrics
583 Views 66 Downloads 649 Total



Related Search

By author names