CLINICAL CYTOGENETIC

Pengaruh Fraksi Jintan Hitam (Nigella Sativa L .) Terhadap Proliferasi dan Apoptosis Pada Sel HeLaZiske Maritska, MD, Msi.Med.Sagita Darma SariFakultas KedokteranUniversitas SriwijayaLaporan Kemajuan Penelitian Dosen Muda SATEKs UnSriIndralaya, Juli 2015

OutlineBackgroundResearch QuestionsMethodsResults

Background

4th most common cancer in women world wide(528,000 new cases in 2012)

4th most common cause of cancer death in women(266,000 deaths in 2012 )The importance of Cervical CancerKim et al., 2011Green et al, 2015http://globocan.iarc.fr/old/FactSheets/cancers/cervix-new.asphttp://www.iarc.fr/en/media-centre/pr/2013/pdfs/pr223_E.pdf

Current treatment of cervical cancerOptions : Surgery, Radiotherapy, Chemotherapy

Radiotherapy recurrence

Chemotherapy Cisplatin (first line drug) ; various and serious adverse effectsHope for a new cureRadiotherapy recurrence ; chemotherapy cisplatinNew cure yang safe, potent dan efektif3Herbal Medicine : where the hope lies

Anti-tumor activity

Anti-oxidant

Less side effects

Less costly WhichHerbs ?Kalonji, Black Cumin, Habat-ul-Sauda, Jinten Hitam

Nigella Sativa L . Ranunculaceae family

Self-reproducing plant with fruit capsule contaning its seeds

Rich historical and religious background

Native to Southern Europe, North Africa, Southwest Asia;cultivated in many countries

Extensive use of its seeds and oil for medicinal use

Gilani, Jabeen, Khan, 2004Salem, 2005Ahmad et al., 2013Nigella Sativa L . ; miracles whithin its seeds

Salem, 2005Ahmad et al., 2013Anti-bacterialAnti-viralAnti-helminthicAnti-fungalAnti-histaminic Anti-inflammatoryAnti-oxidantImmunomodulatorAnti-cancerVarious and many active compounds has been identified

Its most important active compound and of which the pharmacological properties mostly come from thymoquinone

How thymoquinone fights cancer

El-Mahdy et al., 2005Theoretical Scheme

OutlineBackgroundResearch QuestionsMethodsResults

The Biology of Cancer Research Questions

What type of fraction of Nigella Sativa L. that has cytotoxic effect on HeLa Cell ?Does active fraction of Nigella Sativa L. have cytotoxic effect on HeLa Cell ?Does active fraction of Nigella Sativa L. have anti-proliferative effect on HeLa Cell ?Does active fraction of Nigella Sativa L. have apoptosis effect on HeLa Cell ?What active compound does fraction of Nigella Sativa L. contain ? OutlineBackgroundResearch QuestionsMethodsResults

Methods

Experimental studyon cervical cancer cell line (HeLa cell) usingcisplatin, ethanol extract, n-hexan fraction, ethyl acetate fraction & ethanol-water fraction from Nigella Sativa L.

Will be conducted inthe graduate laboratory of Sriwijaya University, Palembang And

Parasitology laboratory of Gadjah Mada University, Jogjakarta

Research Workflow

Extraction, Fractination & Thin Layer Chromatography (TLC)

Extraction, Fractination & Thin Layer Chromatography (TLC)Extraction, Fractination & Thin Layer Chromatography (TLC)Extraction, Fractination & Thin Layer Chromatography (TLC)

Cytotoxic Test, Apoptosis test,Anti-Proliferative test

Cytotoxic test usingMTT Assay :

HeLa Cells are mixed with all the active fractions being studied and cisplatin count the percentage of cells viability

Cytotoxic TestApoptosis Test Using Flow Cytometry

Anti-proliferative Test

Cells are incubated and monitored at 0 hour, 24 hrs, 48 hrs, and 72 hrs (doubling time) by counting the viable cells.

After incubated results reading by using ELISA reader

OutlineBackgroundResearch QuestionsMethodsResults

Results

Results

Results

Results

Results

Results

Financial ReportPenyusunan dan presentasi proposal penelitian Rp. 1.500.000,Ekstraksi dan FraksinasiPembelian Simplisia Jintan Hitam 1 kg Rp. 315.000,-Pembelian Reagen(Alkohol 96%, N-Hexan, dan Acetyl Acetat) Rp. 380.000,-Biaya Laboratorium Biaya Penelitian dan Peminjaman Alat GelasStandar di Program Pasca Sarjana UNSRIRp. 150.000,-Sewa Alat Bagian Parasitologi FK UGMRp. 150.000,-Biaya Kultur Sel Bagian Parasitologi FK UGMRp. 800.000,-Pembacaan Flow CytometriRP. 300.000,-Sel HeLaRp. 4.000.000,-CisplatinRp. 500.000,-Biaya Pemakaian Barang Habis Pakai (BHP)Rp. 1.650.000,-Biaya Pembelian Bahan Milik PenelitiRp. 1.895.000,-Biaya AkomodasiRp. 1.692.300,-Biaya Setor PajakRp. 500.000,-Penyusunan dan presentasi laporan kemajuanRp. 200.000,-TotalRp. 14.032.300,-

Thank YouReferences

http://globocan.iarc.fr/old/FactSheets/cancers/cervix-new.asphttp://www.iarc.fr/en/media-centre/pr/2013/pdfs/pr223_E.pdf Green et al. Concomitant chemotherapy and radiation therapy for cancer of the uterine cervix (review). 2015 The Cochrane Collaboration. Published by John Wiley & Sons, Ltd. Accessed from http://www.bibliotecacochrane.com/pdf/CD002225.pdf Yoon et al. Treatment outcomes of extended-field radiation therapy and the effect of concurrent chemotherapy on uterine cervical cancer with para-aortic lymph node metastasis. Clinical Radiation Oncology. 2015;10:18Gilani, Jabeen, Khan. A Review of Medicinal Uses and Pharmacological Activities of Nigella Sativa. Pakistan Journal of Biological Sciences. 2004;7(4): 441-451Salem. Immunomodulatory and Theurapetic Properties of Nigella Sativa L. Seeds. International Immunopharmacology. 2005;5:1749-1770Ahmad et al. A Review on Theurapetic Potential of Nigella Sativa : A miracle herb. Asian Pasific Journal of Biomedicine. 2013;3(5):337-352S. Zielinski. Henrietta Lacks Immortal Cells. 2010. Available at http://www.smithsonianmag.com/science-nature/Henrietta-Lacks-Immortal-Cells.html