Grape (Vitis vinifera L.), family Vitaceae, is one of the most important plants used extensively in the cosmetics, pharmaceuticals, food sectors, and alternative medicine due to its high content of flavonoids. Leaves of three varieties namely King Ruby, Thompson, and Crimson, were assessed for their cytotoxic effect on the A-549 lung cancer cell line by the in vitro sulforhodamine B (SRB) assay. The results indicate that the tested varieties have weak cytotoxic activity based on cell viability. The King Ruby variety exerted the highest cytotoxic effect, followed by Crimson and Thompson with cell viability % values of 84.158±0.37, 87.613±0.25, and 91.994±0.72, respectively. The preliminary phytochemical screening of the leaves of the three varieties revealed the presence of carbohydrates, sterols/triterpenes, tannins, and flavonoids. Additionally, the determination of the contents of phenolics, flavonoids, and tannins showed that King Ruby was the richest in phenolics, flavonoids, and tannin contents. This motivate us to investigate the petroleum ether extract of King Ruby leaves by GC/MS to identify its phytochemical content.