THE POTENCY OF CRYPTOBRACHYTONE C IN CRYPTOCARYA PULCHRINERVIA LEAVES AS ANTICANCER IN MCF-7 AND T47D BREAST CANCER CELL LINES

• Main Author: Ratnasari, Jujun
• Format: Dissertations
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/80069
• Institution: Institut Teknologi Bandung

Indonesia is one of the countries in the world’s that has highest biodiversity. Many indigenous plants of Indonesia have properties as medicinal plants that have been used traditionally. One of those plants is Cryptocarya, known by the local name as Medang or Huru. Several Cryptocarya species have been reported to have bioactive compounds, and one of these is Cryptocarya pulchrinervia, an indigenous species to the Sumatera, Kalimantan, and Papua regions. One bioactive compound isolated from the leaves of Cryptocarya pulchrinervia is cryptobrachytone C, which showed cytotoxic activity against the murine leukemia P-388 cell line with an IC50 of 3.03 mg/mL (12.71 ?M). However, the accumulation of this compound in the leaves of C. pulchrinervia and the potency of its compound as an anti-cancer agent in breast cancer cell lines, particularly against MCF-7 and T47D breast cancer, have not been reported yet. Therefore, the objective of this research was to investigate the accumulation of C. pulchrinervia in the leaves and the potency of cryptobrachytone C as an anti-cancer in breast cancer MCF-7 and T47D cell lines. The research was conducted in two-parts. The first part is the analysis of the accumulation profile of cryptobrachytone C in leaves of C. pulchrinervia using MALDI-MSI (Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry Imaging) and GC-MS (Gas Chromatography-Mass Spectrometry). The second part is cryptobrachytone C isolation, analysis of the antioxidant using the DPPH (2,2- diphenyl-1-picrylhydrazyl) test and analyzes the cytotoxic activity of the compound against MCF-7 and T47D cell lines using MTT (3-(4,5- dimetilthiazol-2-yl)-2,5- diphenyl-tetrazolium bromide), then analyze the potency of cryptobrachytone C as anti-cancer, using BrdU-ELISA (5-Bromo-2’-deoxyuridine Enzyme-Linked Immunosorbent Assay) for cell proliferation, annexin/PI staining with flow cytometry for the apoptosis test, Boyden Chamber method for migration test and clonogenic test for analyzing the cell capability to make a colony. The results showed that cryptobrachytone C was accumulated with the highest relative intensity in young leaves (in 2nd leaves) compared to the other leaves, based on physiological age. The highest quantitative of cryptobrachytone C was also in young leaves, amounting to 87.07 ± 47.21 mg. Besides that, the antioxidant test showed that cryptobrachytone C has mild antioxidant activity with an AAI index of 0.57. Meanwhile, the MTT test displayed that its cytotoxicity against the MCF-7 cell line with IC50 of 12.94 ± 0.32 ?M but not against the T47D cell line with IC50 of 65.33 ± 2.33 ?M, and it was non-toxic to the normal cell line MRC-5 with IC50 of 122.57 ± 19.84 ?M. The proliferation test indicated that the IC50 of cryptobrachytone C in MCF-7 cell line can inhibit cell proliferation by an amount of 29.33 ± 21.20% after 12-hour treatment, but its IC50 in T47D cell line can inhibit cell proliferation by an amount of 51.42 ± 3.31%. The result of the apoptosis test revealed that the IC50 of cryptobrachytone C in MCF-7 cell line induced early apoptotis 14.75 ± 1.39%, and its IC50 in T47D cell line induced early apoptotis 14.17 ± 2.64%. The migration test revealed that the IC50 of cryptobrachytone C in MCF-7 cell line can inhibit cell migration by 72.41 ± 9.27%, while the IC50 of cryptobrachytone C in T47D cell line can inhibit cell migration 57.69 ± 4.16%. According to clonogenic test results, the IC50 of cryptobrachytone C in MCF-7 cell line can inhibit colony formation with a surviving fraction (SF) of 0.32 ± 0.04 while its IC50 in T47D cell line can inhibit colony formation with SF of 0.55 ± 0.12. From all of the results of the tests, it can be concluded that the accumulation profile of cryptobrachytone C in young leaves of C. pulchrinervia was important information that can be used to gain maximum isolation of cryptobrachytone C from the leaves. This information can also be used in advance research to develop cryptobrachytone C in vitro. Moreover, the cytotoxic of cryptobrachytone C against the MCF-7 cell lines suggests that this compound has potency as an anticancer drug in the MCF-7 cell lines.