Five Gynostemma pentaphyllum (GP) samples were investigated and compared for their chemical compositions and their antioxidant, antiproliferative, and anti-inflammatory effects. Extracts (50% acetone, 75% ethanol, and 100% ethanol) of the five GP samples (GP1-5) differed in their total phenolic, saponin, and flavonoid contents and in their rutin and quercetin concentrations. The highest level of total flavonoids was 63.5 mg of rutin equiv/g in GP4, and the greatest total phenolic content was 44.3 mg of gallic acid equiv/g in GP1 with 50% acetone as the extraction solvent. GP2 had the highest total saponin content of 132.6 mg/g with 100% ethanol as the extraction solvent. These extracts also differed in their scavenging capacity against DPPH and hydroxyl radicals, although they all showed significant radical scavenging capacity. The 100% ethanol extracts also showed dose-dependently strong inhibition on IL-6 and Ptgs2 mRNA expression and weak inhibition on TNF-α mRNA expression. In addition, GP1 had the highest antiproliferative activity at 3.2 mg equiv/mL concentration in HT-29 human colon cancer cells. The results from this study will be used to promote the application of G. pentaphyllum for improving human health.