Cytotoxic effect of γ-sitosterol from Kejibeling (Strobilanthes crispus) and its mechanism of action towards c-myc gene expression and apoptotic pathway
Background: This study aimed to analyze the cytotoxicity effect of γ-sitosterol isolated from “Kejibeling” (Strobilanthes crispus), a medicinal plant, on several cancer cell lines. The mechanisms of the effects were studied through the expression of cancer-caused gene, c-myc and apoptotic pathways.
Methods: This in vitro study was done using human colon cancer cell lines (Caco-2), liver cancer cell lines (HepG2), hormone-dependent breast cancer cell lines (MCF-7) and the normal liver cell lines (Chang Liver). The cytotoxic effect was measured through MTT assay and the potential cytotoxic value was calculated by determining the toxic concentration which may kill up to 50% of the total cell used (IC50). Meanwhile, the cytotoxic mechanism was studied by determining the effect of adding γ-sitosterol to the c-myc gene expression by reverse transciptase-polymerase chain reaction (RT-PCR). The effect of γ-sitosterol through apoptotic pathway was studied by using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.
Results: γ-sitosterol was cytotoxic against Caco-2, HepG2, and MCF-7 with IC50-values of 8.3, 21.8, and 28.8 μg/mL, respectively. There were no IC50-values obtained from this compound against Chang Liver cell line. This compound induced apotosis on Caco-2 and HepG2 cell lines and suppressed the c-myc genes expression in both cells.
Conclusion: γ-sitosterol was cytotoxic against colon and liver cancer cell lines and the effect was mediated by down-regulation of c-myc expression and induction of the apoptotic pathways.
American Cancer Society. Cancer facts and figures 2014. Atlanta : American Cancer Society; 2014.
Wahidin M, Noviani R, Hermawan S, Andriani V, Ardian A, Djarir H. Population-based cancer registration in Indonesia. Asian Pacific J Cancer Prev. 2012;13(4):1709-10. http://dx.doi.org/10.7314/APJCP.2012.13.4.1709
Sunarto PA. Materia Medika Indonesia. Jakarta: Direktorat Jenderal Pengawasan Obat dan Makanan; 1977. Indonesian.
Brummit RK, Powell CE. Authors of plant names. Britain: Royal Botanic Gardens; 1992. p.731.
Kusumoto JT, Shimada I, Kakiuchi N, Hattori M, Namba T. Supriyatna S. Inhibitory effects of Indonesian plant extracts on reverse transcriptase of an RNA tumour virus (I). Phytotherapy Research. 1992;6(5):241-4. http://dx.doi.org/10.1002/ptr.2650060504
Susi E, Asmah R, Patimah I, Taufiq-Yap YH. Comparing of the cytotoxicity properties and mechanism of Lawsonia inermis and Strobilanthes crispus extract against several cancer cell lines. J Med Sci. 2007;7(7):1098-102. http://dx.doi.org/10.3923/jms.2007.1098.1102
Ali AM, Mackeen MM, Intan-Safinar I, Hamid M, Lajis NH, El-Sharkawy SH, et al. Antitumour-promoting and antitumour activities of the crude extract from the leaves of Juniperus chinensis. Journal of Ethnopharmacology. 1996;53(3):165-9. http://dx.doi.org/10.1016/0378-8741(96)01434-1
Chi V. Dang. Myc on the path to cancer. Cell 2012;149(1):22-35. http://dx.doi.org/10.1016/j.cell.2012.03.003
Tuntiwechapikul W, Taka T, Songsomboon C, Kaewtunjai N, Imsumran A, Makonkawkeyoon L, et al. Ginger extract inhibits human telomerase reverse transcriptase and c-Myc expression in A549 lung cancer cells. J Med Food. 2010;13(6):1347-54. http://dx.doi.org/10.1089/jmf.2010.1191
Unger C, Popescu R, Giessrigl B, Rarova L, Herbacek I, Seelinger M, et al. An apolar extract of Critonia morifolia inhibits c-Myc, cyclin D1, Cdc25A, Cdc25B, Cdc25C and Akt and induces apoptosis. Int J Oncol. 2012;40(6):2131-9.
Kolfschoten GM, Hulscher TM, Schrier SM, van Houten VM, Pinedo HM, Boven E. Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin. Gynecol Oncol. 2002;84(3):404-12. http://dx.doi.org/10.1006/gyno.2001.6537
Solary E, Plenchette S, Sordet O, Rébé C, Ducoroy P, Filomenko R, et al. Modulation of apoptotic pathways triggered by cytotoxic agents. Therapie. 2001;56:511-8.
Clary A, Larrue A, Pourquier P, Robert J. Transcriptional down-regulation of c-Myc expression in an erythroleukemic cell line, K562, and its doxorubicin-resistant variant by two topoisomerase II inhibitors, doxorubicin and amsacrine. Anticancer Drugs 1998;9(3):245-54. http://dx.doi.org/10.1097/00001813-199803000-00007
Lane DP. A death in the life of p53. Nature. 1993;362:786-7. http://dx.doi.org/10.1038/362786a0
N, Triaspolitica. "Mengenal Penyakit Kanker, Jenis, Gejala, Penyebab Berikut Pengobatan Kanker." Mau Nanya Dong Dok. N.p, 20 June 2017. Web. 28 June 2017. Retrieved from: https://nanyadongdok.blogspot.com/2017/06/mengenal-penyakit-kangker-jenis-gejala.html
Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991; 139(2):271-9. http://dx.doi.org/10.1016/0022-1759(91)90198-O
Evans DL, Dive C. Effects of cisplatin on the induction of apoptosis in proliferating hepatoma cells and nonproliferating immature thymocytes. Cancer Res. 1993;53(9):2133-9.
Bortner CD, Oldenburg NB, Cidlowski JA. The role of DNA fragmentation in apoptosis. Trends Cell Biol. 1995;5(1):21-6. http://dx.doi.org/10.1016/S0962-8924(00)88932-1
Gold R, Schmied M, Giegerich G, Breitschopf H, Hartung HP, Toyka KV, et al. Differentiation between cellular apoptosis and necrosis by the combined use of in situ tailing and nick translation techniques. Lab Invest. 1994;71(2):219-25.
Copyright (c) 2015 Susi Endrini, Asmah Rahmat, Patimah Ismail, Y.H. Taufiq-Yap
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with Medical Journal of Indonesia agree to the following terms:
- Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
- Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.