Penelitian ini bertujuan untuk meningkatkan zat antikanker dan uji sitotoksitasnya pada T47D dengan menggunakan metode pemupukan dan induksi pada tanaman Curcuma mangga Val. Penelitian dilaksanakan di Greenhouse milik Pusat Inovasi Agroteknologi Terpadu (PIAT) Universitas Gadjah Mada di Kalitirto, Kecamatan Berbah, Sleman, Yogyakarta. Waktu pelaksanaan awal Januari-Oktober 2019. Pelaksanaan penelitian dibagi menjadi 3 tahap. Tahap pertama penanaman dan pemupukan C. mangga Val. Pemupukan dilakukan menggunakan nitrogren (N) organik dari kompos daun-daun legume, fosfor (P) organik dari pupuk SP36, Kalium (K) dari KCl, dan kombinasi pupuk NPK. Tahap kedua adalah ekstraksi dan penetapan kandungan zat aktif menggunakan alat vaccum-rotavapor dan densitometer. Tahap ketigaadalah Uji sitotoksisitas (MTT Assay) menggunakan bahan Sel T47D, media RPMI, MTT, PBS, SDS, dan bahan-bahan disposable, serta ELISA reader. Hasil penelitian menunjukan bahwa pemupukan N organik pada C. mangga Val. mampu meningkatkan produksi minyak dalam rimpang dan sitotoksisitas minyak terhadap sel kanker payudara T47D. Penghentian pemberian air menjelang masa dorman dan suhu lingkungan yang panas (>37oC) mampu menginduksimajunya masa dorman pada tanaman C. mangga Val.INCREASING ANTICANKER SUBSTANCES AND THE Cytotoxicity TEST ON T47DUSING FERTILIZATION AND INDUCTION ON Curcuma Mango Val.This study was aimed at increasing the anticancer agent and conducting a cytotoxicity test on T47D by using fertilization and induction methods on Curcuma Val mango plant. The study was conducted at the Greenhouse of Agro-technology Innovation Center (AIC), Gadjah Mada University in Kalitirto, Berbah, Sleman, Yogyakarta. The study was conducted from January to October 2019. The research was divided into 3 stages. The fi rst stage of planting and fertilizing C. mango Val. Fertilization was carried out using organic nitrogren (N) fromcompost of legume leaves, organic phosphorus (P) from SP36 fertilizer, Potassium (K) from KCl and NPK fertilizer combination. The second step is the extraction and the determination of the active ingredient using a vacuum-rotary evaporator and a densitometer. The third stage is the cytotoxicity test (MTT Assay) using T47D cell material, RPMI media, MTT, PBS, SDS and disposable materials, and ELISA reader. The results showed that organic N fertilizationon C. mango Val. able to increase oil production in rhizomes and cytotoxicity of oil against T47D breast cancer cells. Discontinuation of water delivery prior to dormancy and hot ambient temperature (> 37oC) is able to induce the advancement of dormancy in C. mango Val plants.
Semisynthesis of D6,7-Anhydroerythromycin-A was done by biomodification technique by addition of 0.2% INH into a culture fermentation of Saccharopolyspora erythraea ATCC 11635 in medium Hutchinson. The aim of this research is to studies of fragmentation pattern from new matabolite of D6,7-Anhydroerythromycin-A by Liquid Chromatography-Mass Spectroscopy (LC-MS) and the ionization of mass spectroscopy is use by ESI (Electrospray Ionization) pattern. The FT-IR spectrometric analyzes showed a stretching vibration of C=C conjugated group at wave number 1602.7 cm-1. This C=C conjugated vibration indicated the existence of double bond between C6 and C7 (D6,7), this confirmed that isolate contained D6,7-Anhydroerythromycin-A (the possibility of D6,7 was positive). For complementation, a LC-MS (Liquid Chromatography-Mass Spectroscopy) analyzes using ESI-MS (Electrospray Ionization-Mass Spectroscopy) ionization pattern was conducted to the isolate which resulted Quassimolecular ions [M+H]+ of D7,8- and D6,7-Anhydroerythromycin-A. LC-MS spectrogram of the isolate, which gave two peaks of m/z 732.2460 and m/z 716.2522, confirmed that the m/z 732.2460 possibly was D7,8-Anhydroerythromycin-A, while the m/z 716.2502 and m/z 715.2522 possibly were D6,7-Anhydroerythromycin-A. Keywords: isoniazid, enoyl reduction, D6,7-Anhidroeritromisin-A, fragmentation, LC-MS.
Indonesia memiliki sekitar 200 jenis bajakah; 4 di antaranya yaitu bajakah tampala (Spatholobus littoralis Hassk.), kalalawit (Uncaria gambir (Hunter) Roxb.), bajakah jari lima (Schefflera petiolosa (Miq.) Harms), dan longkur (Spatholobus ferrugineus (Zoll. & Moritzi) Benth.) yang telah digunakan oleh suku Dayak untuk pengobatan kanker payudara. Studi ini menguji sitotoksisitas antikanker payudara ekstrak akar dari keempat bajakah di atas. Ekstraksi menggunakan sample grinding pestle dengan pelarut air, air panas, etanol 96%, dan n-heksan. Uji sitotoksisitas ekstrak dilakukan secara in vitro terhadap sel T47D dengan pembanding tamoksifen. Nilai IC50 dari uji sitotoksik ekstrak etanol, air panas, air, dan n-heksan berturut-turut: 1136; 1871,5; 2294,25; dan 3975,5 µg/mL (R2 = 0,962; 0,943; 0,943; dan 0,956), tamoksifen 9 µg/mL (R2 = 0,9581). Ekstrak etanol bajakah merupakan yang paling toksik dibanding lainnya. Sehingga untuk perbaikan nilai IC50 dilakukan ekstraksi reflux menggunakan etanol 96% pada keempat akar bajakah. Hasil menunjukkan nilai IC50 bajakah kalalawit, tampala, longkur, dan jari lima yang membaik yaitu berturut-turut adalah 407; 708; 881; dan 1.096 µg/mL (R2 = 0,9717; 0,952; 0,9367; 0,9369). Keempat ekstrak bajakah mengandung zat aktif antikanker payudara (mengingat nilai R2 uji sitotoksisitas > 0,93), namun kadar yang terekstraksi sangat kecil sehingga perlu dilakukan ekstraksi dengan menggunakan soxhlet.
C. mangga Val. has been used as an alternative remedy for cancer in Yogyakarta. The protein fraction of C. mangga was identified to contain Ribosome-inactivating Protein which cleave supercoiled double stranded DNA in vitro. In this experiment, the protein fraction isolated from fresh, 400C dried and freeze dried C. mangga Val. rhizome was screened against HeLa, Raji cell-lines and normal mononuklear cells for cytotoxic effects. This would enable us to describe the sensitivity of the protein extract on different cell types. The level of cytotoxicity was determined on the level of LC50 which was based on the percentage of the cell death following the 24 hours incubation with the extract. The protein isolated from C. mangga Val. was able to cleave supercoiled double stranded DNA to nick circular form. This result suggested that protein contained RIPs. The highest activity was identified in the protein isolated from fresh C. mangga Val, and this was followed respectively by freeze drying and 400C drying C. mangga Val. The comparison of the cytotoxic effect showed that protein of fresh C. mangga Val produced the largest number of death cells and the most toxic was on the HeLa cell line. Moreover, the LC50 indicated that the highest cytotoxic effect was shown by protein isolated from fresh C. mangga Val. followed respectively by freeze drying and 400C drying C. mangga Val. Based on LC50, the highest cytotoxic effect of C. mangga Val was found on HeLa cell line, while similar cytotoxic effect was appear on Raji cell line and normal mononuclear cells Keywords: C. mangga Val., RIP, cytotoxic, cancer cell lines, normal cell
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