nan Sismindari

Mekanisme In Vitro Induksi Apoptosis dari RIP (Ribosome- Inactivating Protein) Daun Bunga Pukul Empat (Mirabilis jalapa L) In Vitro Mechanism of Apoptosis Induction of RIP (Ribosome- InactivatingProtein) Isolated from Bunga Pukul Empat (Mirabilis jalapa L

ABSTRACT Background: Ribosome-inactivating protein (RIP) isolated from the leave of Mirabilis jalapa L (MJ Protein) has shown to possess anticancer activity by triggering apoptosis in HeLa cells in vitro. This study aimed at finding out the trigger mechanisms that induces apoptosis of RIP. Design and Method: This study was conducted by giving the treatment of MJ Protein to the Hela cell culture in vitro. Then the expression of and activation of caspase-3 were observed using Immunohistochemistry and colorimetry method. Result: The data showed that the MJ protein treatment increased the caspase-3 expression and increased the activity of caspase-3 in Hela cel culture in vitro. Conclusion: This result indicates the anticancer potential of MJ protein through its effect on signal tranduction process leading to apoptosis through caspase activation with caspase-3 as apoptosis executor (Sains Medika, 2(1): 23-31). Key words: Mirabilis jalapa L , Protein MJ, apoptosis, caspase-3 ABSTRAK Pendahuluan: Ribosome-inactivating protein (RIP) dari daun Mirabilis jalapa, L (Protein MJ) telah diketahui potensinya sebagai antikanker karena mampu memacu apoptosis secara in vitro pada kultur sel HeLa. Penelitian ini dilakukan untuk mengetahui kemungkinan jalur mekanisme pemacuan apoptosis dari RIP tersebut, apakah melalui aktivasi caspase. Metode Penelitian: Penelitian ini dilakukan dengan memberikan perlakuan Protein MJ secara in vitro pada kultur sel HeLa, kemudian dilihat ekspresi dan aktivitas caspase-3 melalui pengecatan imunositokimia dan dengan metode kolorimetri. Hasil penelitian: Data penelitian menunjukkan bahwa pemberian Protein MJ mampu meningkatkan ekspresi caspase-3 dan meningkatkan aktivitas caspase-3 pada kultur sel HeLa secara in vitro. Kesimpulan: Hasil penelitian tersebut mengimplikasikan potensi antikanker dari Protein MJ melalui pengaruhnya pada proses transduksi sinyal yang memacu apoptosis, melalui aktivasi caspase, dengan caspase-3 sebagai eksekutor terjadinya apoptosis (Sains Medika, 2 (1): 23-31).

CITOTOXICITY TEST OF THE PROTEIN FRACTIONS OF CARICA PAPAYA L. LEAVES ON HeLa CELL LINE.

Previous research showed that a RIP like protein from Carica papaya L. leaves was available on 60 – 80% saturation ammonium sulfate fraction. In this research cytotoxicity assay of the protein fractions was carried out on HeLa cell lines. RIP-like protein from Carica papaya L. leaves, isolated leg ammonium sulfat saturation was prepared based on the ability of cleaving double stranded supercoiled DNA. This fraction was then tested its cytotoxicity on HeLa cell lines. The results indicated that fraction at 60% and 80% saturation possessed lower activity with the LC50 of 1,35 mg/ml and 0,936 mg/ml, respectively compared to the crude extracts (LC50 0,623 mg/ml). The higher toxicity level of the crude extract might be caused also by toxicity of other compounds containing in the crude extract. Key word : cytotoxicity, protein fraction, HeLa cell line