Perbandingan Uji Aktivitas Sitotoksik In Vitro dan In Silico Antikanker Payudara Senyawa Ekstrak Heksan dan Minyak Atsiri Rimpang Curcuma mangga Val.

PUTRI KHAERANI C, Prof. Dr. apt. Retno Sunarminingsih S., M.Sc.; Dr. apt. Purwanto, M.Sc.

2021 | Tesis | MAGISTER ILMU FARMASI

Sitotoksisitas ekstrak dan minyak atsiri rimpang Curcuma mangga Val. telah terbukti berpotensi sebagai anti kanker payudara. Secara in silico, kandungan minyak atsiri C. mangga Val. terbukti sebagai inhibitor reseptor ER-α. Tetapi belum ada peneliti yang membandingkan sitotoksisitas ekstrak dan minyak atsirinya secara in vitro terhadap T-47D dan MCF-7; serta afinitas senyawa aktif dalam ekstrak dan minyak atsiri tersebut terhadap protein reseptor ER-α. Oleh karena itu, penelitian ini bertujuan untuk membandingkan sitotoksisitas ekstrak n-heksan dan minyak atsiri terhadap sel T-47D dan MCF-7, serta afinitas beberapa senyawa aktif yang terkandung dalam sampel terhadap reseptor ER-α secara in silico. Senyawa aktif rimpang C. mangga Val. (segar maupun kering) dimaserasi menggunakan n-heksan dan di-destilasi uap air untuk mendapatkan ekstrak n-heksan dan minyak atsiri. Selanjutnya, semua sampel diuji sitotoksisitasnya secara MTT assay terhadap sel T-47D dan MCF-7. Kandungan sampel diidentifikasi menggunakan GC-MS. Ligan molecular docking diseleksi berdasarkan similarity index (SI) > 850 dan kelimpahan relatif >1% untuk menentukan docking score terhadap protein reseptor ER-α dengan ligan antagonis 4-hidroksi tamoxifen serta ligan agonis estradiol, estriol dan estron; menggunakan metode MOE 2015.10. Uji sitotoksisitas menunjukkan ekstrak n-heksan rimpang segar C. mangga Val. memiliki potensi sitotoksik terbaik dengan IC50 36,308 µg/ml terhadap T-47D; dan 68,077 µg/ml terhadap sel MCF-7. Identifikasi menggunakan GC-MS memperlihatkan perbedaan kandungan senyawa terbesar pada ekstrak n-heksan dan minyak atsiri rimpang C. mangga Val.; yaitu dalam ekstrak adalah golongan diterpenoid dengan senyawa (E)-labda-8(17),12-diene-15,16-dial (rimpang kering 41,61%; rimpang segar 83,84%); sedangkan dalam minyak atsiri adalah golongan monoterpenoid dengan senyawa β-myrcene (rimpang kering 18,30%; rimpang segar 26,28%). Delapan belas ligan uji terseleksi dari hasil identifikasi kandungan ekstrak n-heksan dan 26 ligan uji dari minyak atsiri. Docking score senyawa ekstrak n-heksan dan minyak atsiri yang paling baik dibanding skor estradiol (-6,1074) adalah m-Camphorene (-7,3390). Isolat ekstrak n-heksan lebih sitotoksik terhadap sel kanker payudara daripada minyak atsiri rimpang C. mangga Val.

Cytotoxicity studies of the extract and essential oil Curcuma mangga Val. rhizomes showed they are potential as anti-breast cancer. In silico studies proved that the active compounds of the essential oil were inhibitor of ERα. There is no researcher has compared the cytotoxicity of the extract and essential oil towards T-47D and MCF; and also the affinity of the active compounds contained in the extract and essential oil towards ERα. So that, this study is to compare the cytotoxicity against T-47D and MCF-7 cells of the n-hexane extract and essential oil of C. mangga Val. rhizomes, as well as in silico affinity of the active compounds contained in the samples towards ER-α. Compounds of the C. mangga Val. rhizomes (either fresh or dry) were n-hexane macerated and water steam distillated to get the n-hexane extract and essential oil. All samples were then cytotoxicity tested using MTT assay towards T-47D and MCF-7. The samples compounds were identified using GC-MS. Compounds with similarity index (SI) > 850 and relative area >1% then were used for molecular docking studies using MOE 2015.10 to determine the docking score towards ERα with antagonist ligand 4-hydroxy tamoxifen and agonist ligands of estradiol, estriol dan estron. Cytotoxicity studies showed that the fresh-rhizome-extract of C. mangga Val. was the most cytotoxic with IC50 36,308 µg/ml towards T-47D and 68,077 µg/ml towards MCF-7. GC-MS identification showed different biggest contents between the n-hexane extract and its essential oil of C. mangga Val. rhizomes; which was a diterpenoid of E)-labda-8(17),12-diene-15,16-dial (from the dry-rhizomes 41,61%; fresh-rhizomes 83,84%) in the extract; while in the essential oil was a monoterpenoid of β-myrcene (from the dry-rhizomes 18,30%; fresh-rhizomes 26,28%). Eighteen substances were selected from the identification of the n-hexane extract contents, and 26 substances were selected from the essential oil to became the test ligands. The best docking score of the n-hexane extract and the essential oil substances comparing to that of estradiol (-6,1074) was m-Camphorene (-7,3390). Isolate n-hexane extract was more cytotoxic towards breast cancer cells comparing to the essential oil of the C. mangga Val. rhizome.

Kata Kunci : Curcuma mangga Val., terpenoid, T-47D, MCF-7, ER-α