IDENTIFIKASI GOLONGAN SENYAWA POTENSIAL PENGHAMBAT PERTUMBUHAN SEL KANKER PAYUDARA (T47D) ORGAN VEGETATIF Dendrobium lasianthera J.J. Sm

Sri Handayani, Dr. L. Hartanto Nugroho, M.Agr.Sc.

2012 | Tesis | S2 Biologi

Dendrobium lasianthera J.J. Sm memiliki kedekatan kekerabatan dengan anggota marga Dendrobium yang telah diketahui memiliki efek farmakologis karena kandungan senyawa bioaktifnya. Penelitian tentang kandungan dan manfaat senyawa bioaktif pada D. lasianthera J.J. Sm belum pernah dilakukan, untuk itu dilakukan penelitian tentang Identifikasi Golongan Senyawa Potensial Penghambat Sel Kanker Payudara (T47D) Organ Vegetatif Dendrobium lasianthera J.J. Sm D. lasianthera J.J. Sm yang digunakan berasal dari Vanindita Nursery, Wedomartani, Ngemplak, Sleman. Organ vegetatif yang digunakan berupa akar, batang dan daun yang dimaserasi menggunakan tiga macam pelarut yaitu kloroform, metanol dan air. Ekstrak diujikan pada sel kanker payudara (T47D) dengan metode 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. Ekstrak potensial difraksinasi dengan metode Vacuum Liquid Chromatography (VLC), selanjutnya fraksi yang bersifat toksik difraksinasi kembali dengan Kromatografi Lapis Tipis preparatif (KLT preparatif). Golongan senyawa bioaktif diidentifikasi dengan Kromatografi Lapis Tipis (KLT) menggunakan silika gel GF254 yang disemprot dengan reagen serium (IV) sulfat, liebeman-burchard, vanillin sulfat, dragendorff dan amonium serta diamati di bawah lampu Ultra Violet (UV) λ 254 dan 366 nm. Analisis data untuk menentukan nilai Inhibitory Concentration of 50% (IC 50) menggunakan persamaan regresi linier. Nilai IC50 selanjutnya dianalisis statistik menggunakan Analysis Of Variance) ANOVA dan hasil yang berbeda secara signifikan dilanjutkan dengan Duncan’s Multiple Rate Test (DMRT). Hasil penelitian menunjukkan bahwa ekstrak kloroform batang D. lasianthera J.J. Sm memiliki nilai IC50 paling rendah, yaitu 117 µg/ml. Ekstrak potensial dipartisi dan diuji sitotoksisitasnya sehingga diperoleh fraksi gabungan toksik, yaitu fraksi yang terdiri atas senyawa kurang polar dengan nilai IC50 sebesar 122,5 µg/ml. Fraksi gabungan toksik dipisahkan sehingga dihasilkan fraksi KLT preparatif. Uji sitotoksisitas fraksi KLT preparatif menghasilkan fraksi yang bersifat paling toksik dengan nilai IC50 sebesar 37,03 µg/ml. Nilai IC50 fraksi KLT preparatif toksik lebih rendah dibandingkan dengan nilai IC50 obat kanker doxorubicine (61,67 µg/ml). Hasil identifikasi fraksi KLT preparatif toksik menunjukkan bahwa senyawa potensial penghambat pertumbuhan sel kanker payudara (T47D) adalah golongan terpenoid dan fenol.

Dendrobium lasianthera J.J. Sm is a member of genus Dendrobium that have been known having pharmacological effects due to its bioactive compounds. There is no scientific justification about the contents and the benefits of D. lasianthera J.J. Sm bioactive compounds. Therefore, the current research is aimed to identify the potential compound for breast cancer cells (T47D) growth inhibitor from vegetative organs of Dendrobium lasianthera J.J. Sm. D. lasianthera J.J. Sm were collected from Vanindita Nursery, Wedomartani, Ngemplak, Sleman. The materials used were roots, stems and leaves. The materials were macerated using three different solvents, namely chloroform, methanol and water. The extracts were then tested for their toxicity on breast cancer cells (T47D) using -(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. Potential extracts were fractionated by Vacuum Liquid Chromatography (VLC) method. Potential fraction were isolated using preparative Thin Layer Chromatography (preparative TLC) and the compound was identified by Thin Layer Chromatography (TLC) using silica gel GF254 and sprayed using cerium (IV) sulfate, liebeman-burchard, vanillin sulfate, dragendorff and ammonium, and then was observed under Ultra Violet (UV) light λ 254 and 366 nm. The determination of Inhibitory Concentration of 50% (IC 50) was calculated using linear regression equation. Then IC50 value was test with Analysis Of Variance (ANOVA) and were then test with Duncan’s Multiple Rate Test (DMRT) for their significantly different. The results showed that the chloroform extract of D. lasianthera J.J. Sm stem had the lowest IC50 value of 117 µg/ml compared to other extracts. The extract were partitioned and tested to obtain the potential cytotoxicity of the fraction. The fraction that have low polarity was the most toxic fraction with IC 50 value of 122.5 µg/ml. The low polarity fraction was then separated again. The preparative TLC fraction were tested on cytotoxicity assay in order to obtain the most toxic fraction. The most toxic preparative TLC fraction had IC 50 value of 37.03 µg/ml. The IC50 value of the most toxic preparative TLC fraction was lower than the cancer drug, doxorubicine, which had the IC50 value of 61.67 µg/ml. The identification of the most toxic preparative TLC fraction showed that the potential growth inhibitor of breast cancer cells (T47D) compounds were terpenoids and phenols.

Kata Kunci : Dendrobium lasianthera J.J. Sm, senyawa bioaktif, organ vegetatif, antikanker.