Regulation of Chloride Intracellular Channel Protein 1 and Caspase -3 mRNA Expression by Hydroethanolic Extract of Aegle marmelos Fruit Human Breast Cancer Cell Line-MCF-7

Abstract

Introduction: Cancer is the second leading cause of death all over the world where among all types of cancer breast cancer is said to be the leading cancer followed by lung cancer. The aim of this study is to find the regulation of chloride intracellular channel protein 1 and caspase -3 mRNA expression by hydroethanolic extract of Aegle marmelos fruit human breast cancer cell line-MCF-7. Materials and methods: MCF-7 cells were collected from NCCS Pune, India. It is stored in Dubecos Modified Eagle's Medium. The Aegle marmelos fruit was collected from the herbal department and its extract was prepared. The extract of Aegle marmelosis used in treating MCF-7 cells at different dosages in in vitro.  Isolation of total RNA from MCF-7 cells. The cells will be mixed with total RNA isolation reagent, sonicated and RNA will be isolated as per the standard method. c-DNA conversion and real time polymerase chain reaction. The c-DNA will be synthesized using reverse transcription by commercially available (RT-PCR) kit. Two microlitres of c-DNA will be used for amplification of clic-1 and caspase-3 using gene specific primers by commercially available RT-PCR kit (SyBr kit) and comparative CT method will be used to see the expression of genes. Untreated MCF-7 cells were compared with MCF-7 cells treated with various concentrations of the extract (10, 20 and 40ug). The statistical data’s were collected from the SPSS software version 21. Result: The given extract inhibits the proliferation of MCF-7 cells therefore said to have antiproliferative activity. Different doses of extract were tested (200ug-500ug) out of which 400ug of extract were preferred. Conclusion: The given plant extract has anti proliferative properties and hence can be used as a drug to treat breast cancer.