EFFECTIVENESS EXTRACT AIR purwaceng (Pimpinella alpina) INDUCTION OF APOPTOSIS AND APOPTOSIS GENE EXPRESSION PROTEIN IN VARIOUS CELL LUNG CANCER A549
ABSTRACT: Background: Lung cancer is the 3rd most frequent occurring cancer and the leading cause of cancer related death in men. Conventional treatment of lung cancer consists of multidrug chemotherapy. Although chemotherapy regiments has been proven to be effective, there are many highly undesirable side effects. Thus, an alternative chemotherapy agent is needed that has similar efficacy of conventional chemotherapy with minimal side effects. Alternative chemotherapy agent is an agent that has cytotoxic activity through the mechanism of cell cycle arrest, apoptosis induction capability, or trigger the expression of proteins that are able to induce apoptosis. Purwaceng (Pimpinella alpina) has Coumarin active compound that is known to have chemotoxic effect towards cancer cells. Role of Purwaceng (Pimpinella alpina) on the cytotoxic effect and induced apoptosis in lung cancer cells remains unclear. Objective: To determine the cytotoxic effects and apoptosis induction ability of water extracts of Pimpinella alpina for A549 lung cancer cells through apoptosis protein expression of various genes. Method: The study is purely experimental that used A549 lung cancer cells that were given a Purwaceng extract dose range of 1-10.000 μg/mL with 24-48 hours of incubation time. For comparison, a positive control containing cisplatin was used with a dose range of 1- 500 μg/mL. Cytotoxicity study were stained using MTT, where as apoptosis induction study used ethidium orange-acridine bromide. Protein expression of apoptosis induction study using anti-p53, Rb, Bcl-2, and Caspase-9 antibody immunohistochemitry staining. Result: The cytotoxicity results show incubation for 24 hours resulted in an IC50 of 3.862,00 μg/mL. Apoptosis induction study shows an EC50 value of 5.271,00 μg/mL, identified by the decrease in Rb and Bcl-2 expression and increase in p53 and caspase-9 expression. Chisquare statistical analysis shows proportionate effects of IC50 towards gene expression that are statistically significant (p<0.05) in comparison to negative control. Conclusion: This study shows Pimpinella alpina extract has cytotoxic and apoptosis inductive properties towards A549 lung cancer cell, identified by the decrease in Rb and Bcl-2 expression and increase in p53 and caspase-9 expression.
ABSTRACT: Background: Lung cancer is the 3rd most frequent occurring cancer and the leading cause of cancer related death in men. Conventional treatment of lung cancer consists of multidrug chemotherapy. Although chemotherapy regiments has been proven to be effective, there are many highly undesirable side effects. Thus, an alternative chemotherapy agent is needed that has similar efficacy of conventional chemotherapy with minimal side effects. Alternative chemotherapy agent is an agent that has cytotoxic activity through the mechanism of cell cycle arrest, apoptosis induction capability, or trigger the expression of proteins that are able to induce apoptosis. Purwaceng (Pimpinella alpina) has Coumarin active compound that is known to have chemotoxic effect towards cancer cells. Role of Purwaceng (Pimpinella alpina) on the cytotoxic effect and induced apoptosis in lung cancer cells remains unclear. Objective: To determine the cytotoxic effects and apoptosis induction ability of water extracts of Pimpinella alpina for A549 lung cancer cells through apoptosis protein expression of various genes. Method: The study is purely experimental that used A549 lung cancer cells that were given a Purwaceng extract dose range of 1-10.000 μg/mL with 24-48 hours of incubation time. For comparison, a positive control containing cisplatin was used with a dose range of 1- 500 μg/mL. Cytotoxicity study were stained using MTT, where as apoptosis induction study used ethidium orange-acridine bromide. Protein expression of apoptosis induction study using anti-p53, Rb, Bcl-2, and Caspase-9 antibody immunohistochemitry staining. Result: The cytotoxicity results show incubation for 24 hours resulted in an IC50 of 3.862,00 μg/mL. Apoptosis induction study shows an EC50 value of 5.271,00 μg/mL, identified by the decrease in Rb and Bcl-2 expression and increase in p53 and caspase-9 expression. Chisquare statistical analysis shows proportionate effects of IC50 towards gene expression that are statistically significant (p<0.05) in comparison to negative control. Conclusion: This study shows Pimpinella alpina extract has cytotoxic and apoptosis inductive properties towards A549 lung cancer cell, identified by the decrease in Rb and Bcl-2 expression and increase in p53 and caspase-9 expression.
