PERAN FRAKSI AKTIF TANAMAN SARANG SEMUT (Myrmecodia pendens Merr & Perry) TERHADAP SEL ORAL BURKITTS LYMPHOMA (Kajian Sitotoksisitas, Sinergitas Ko-kemoterapi Cisplatin, Hambatan Signal Transduksi NF-kB, Komplek CDK-2-cyclin-E, dan Induksi Apoptosis)

ANA MEDAWATI, Drg. Supriatno, MDSc, M.Kes, PhD. ; Prof.dr. Sofia Mubarika, M.Med.Sc, PhD. ; Drh. SitarinaWidyarini, MP,PhD.

2019 | Disertasi | DOKTOR ILMU KEDOKTERAN GIGI

Latar belakang :Usaha menghambat pertumbuhan sel kanker oral Burkitts lymphoma memerlukan strategi yang efektif dan potensial salah satunya menggunakan tanaman sarang semut (Myrmecodia pendens Merr & Perry) yang merupakan tanaman obat yang potensi antitumornya perlu terus diuji secara ilmiah. Tujuan penelitian : Menguji fraksi aktif M. pendens terhadap sel oral Burkitts lymphoma: kajian sitotoksisitas dan sinergitas ko-kemoterapi dengan cisplatin, hambatan signal transduksi komplek CDK-2-cyclin-E, NF-kB, serta induksi apoptosis. Metode: Jenis penelitian yang digunakan adalah eksperimental murni laboratorik dengan rancangan post-test only control group design. Uji sitotoksisitas dilakukan dengan uji MTT (3-(4,5 dimethyl thiazol-2-yl)-2.5-diphenyltetrazolium bromide). Uji sinergitas ko-kemoterapi cisplatin dilaksanakan dengan program CompuSyn. Uji apoptosis diobservasi menggunakan FITC-Annexin V, Induksi apoptosis caspase-3,-8 dan -9 dan mekanisme hambatan pertumbuhan sel pada komplek CDK-2-cyclin-E, dan NF-kB. menggunakan uji kolorimetri dengan ELISA kit. Hasil : Fraksi etil asetat M. pendens memiliki aktivitas sitotoksik paling kuat dengan nilai IC50 sebesar 278,21 mikrogram/ml. Ko-kemoterapi cisplatin dengan fraksi etil asetat M. pendens menunjukkan aktivitas sitotoksik kuat dan sinergitas positif pada level ringan sampai sedang dengan nilai Combination Index (CI) sebesar 0,7-0,9. Fraksi etil asetat M. pendens menunjukkan hambatan pertumbuhan sel oral Burkitts lymphoma melalui mekanisme hambatan signal transduksi NF-kB dan komplek protein CDK2-cyclin-E, serta mempunyai potensi menghambat siklus sel sebagian besar pada fase G0-G1 dan sebagian kecil pada fase G2-M. Hasil uji apoptosis menggunakan uji kolorimetrik caspases dan FITC-Annexin V menunjukkan induksi apoptosis caspase-3,-8 dan -9. Kesimpulan: Ko-kemoterapi cisplatin dengan fraksi etil asetat M. pendens memiliki aktivitas sitotoksisitas dan sinergitas yang positip, potensi hambatan yang kuat terhadap sel oral Burkitts lymphoma melalui mekanisme hambatan siklus sel, penurunan regulasi signal transduksi komplek CDK-2-cyclin-E, NF-kB, dan induksi apoptosis.

Background: The effort to inhibit the growth of Burkitts lymphoma oral cancer cell needs an effective and potential strategy. One of the strategy was using Myrmecodia plant (Myrmecodia pendens Merr & Perr), a medicinal plant which its antitumor properties have been empirically and scientifically tested. Aim: The aim of this study was to test the active fractions of M. pendens towards Burkitts lymphoma oral cell. It was also to study the cytotoxicity and co- chemotherapy synergism with cisplatin, the inhibition of transduction signal of CDK- 2-cyclin-E complex and NK-kB complex, and also the induction of apoptosis. Method: The study was a true laboratory experimental research. The design of this study was post-test only control group design. The cytotoxicity assay was taken with MTT 3-(4,5 dimethyl thiazol-2-yl)-2.5-diphenyltetrazolium bromide) assay. The synergism and co-chemotherapy test with cisplatin was taken with CompuSyn program. The apoptosis was observed with FITC-Annexin V and caspase-3, -8 and -9 assay. Next, the inhibition of cells growth mechanism was detected by the expression assay of CDK-2-cyclin-E complex and NF-kB complex with ELISA kit. Result: The ethyl-acetate fraction of M. pendens had the strongest cytotoxic properties with IC50 value of 278,21 mikrogram/ml. The combination of ethyl-acetate fraction of M. pendens co-chemotherapy with cisplatin exhibited a strong cytotoxic properties and positive synergism with a low to moderate level, with Combination Index (CI) score of 0,7-0,9. The ethyl-acetate fraction of M. pendens revealed the inhibited growth of Burkitts lymphoma oral cell through the inhibition of the transduction signal of NF-KB and CDK2-cyclin-Es protein. It was also noted that it had a potential inhibition activity to cells cycle, mainly during G0-G1 phase and lesser during G2-M phase. The apoptosis test with caspase colorimetric assay and FITC-Annexin V assay showed the induction of proteolytic activity of caspase-3, -8 and -9. Conclusion: The ethyl-acetate fraction of M. pendens had a cytotoxicity activity and co- chemotherapy synergism with cisplatin. It also had a potential inhibition activity of Burkitts lymphomas oral cell, cell cycle arrest, by lowering the transduction signal of CDK-2-cyclin- E complex and NF-κB complex, and it also induced apoptosis.

Kata Kunci : Myrmecodia. pendens, Burkitts lymphoma , sitotoksisitas, ko- kemoterapi, CDK-2-cyclin-E, NF-kB, apoptosis, caspase, Myrmecodia pendens, Burkitts lymphoma, cytotoxicity, co- chemotheraphy, CDK-2-cyclin-E, NF-kB, apoptosis, caspase.