Objective: The current study was aimed to evaluate the methanolic extract of caryopses of Echinochloa colona (ECME) for its in vitro hepatoprotective activity against ethanol in HepG2 cell lines.Methods: In this regard, the cytotoxicity studies were conducted for the extract, ECME using 3-(4,5-dimethythiazol- 2-yl)-2,5-diphenyl tetrazolium bromide assay to determine the inhibitory concentration 50% value based on which, the doses 50, 100, and 200 μg/ml were selected for the hepatoprotective studies in HepG2 cell lines. The toxicity was induced using ethanol (100 mM). The in vitro hepatoprotective activity of the extract was assessed based on the changes in the level of biochemical parameters such as aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase.Results: The extract, ECME has shown a dose-dependent cytoprotective activity with maximum protection at 200 μg/ml. The percentage cell viability of the extract, ECME at 200 μg/ml was more, i.e., 69.33% which was well comparable to that of standard drug, silymarin (100 μg/ml).Conclusion: The study revealed that the extract had shown significant hepatoprotective activity at all the test doses against ethanol-induced cytotoxity assay.