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Fungal endophytes have remarkable potential to produce bioactive compounds with numerous pharmacological significance that are used in various disease management and human welfare. In the current study, a total of eight fungal endophytes were isolated from the leaf tissue of <i>Amoora rohituka</i>, and out of which ethyl acetate (EA) extract of <i>Penicillium oxalicum</i> was found to exhibit potential antioxidant activity against DPPH, nitric oxide, superoxide anion and hydroxyl free radicals with EC₅₀ values of 178.30 ± 1.446, 75.79 ± 0.692, 169.28 ± 0.402 and 126.12 ± 0.636 µg/mL, respectively. The significant antioxidant activity of EA extract of <i>P. oxalicum</i> is validated through highest phenolic and flavonoid content, and the presence of unique bioactive components observed through high-performance thin layer chromatography (HPTLC) fingerprinting. Moreover, EA extract of <i>P. oxalicum</i> also displayed substantial anti-proliferative activity with IC₅₀ values of 56.81 ± 0.617, 37.24 ± 1.26 and 260.627 ± 5.415 µg/mL against three cancer cells HuT-78, MDA-MB-231 and MCF-7, respectively. Furthermore, comparative HPTLC fingerprint analysis and antioxidant activity of <i>P. oxalicum</i> revealed that fungal endophyte <i>P. oxalicum</i> produces bioactive compounds in a host-dependent manner. Therefore, the present study signifies that fungal endophyte <i>P. oxalicum</i> associated with the leaf of <i>A. rohituka</i> could be a potential source of bioactive compounds with antioxidant and anticancer activity.