Very long chain polyunsaturated fatty acids (VLCPUFAs) are widely used as nutraceutical supplements for human health. Recently, a high level of two new VLCPUFAs, docosadienoic acid (DDA, 22:2n-6) and docosatrienoic acid (DTA, 22:3n-3), was produced in oilseed crop Brassica carinata using a biotechnology approach. This study investigated the functional properties of these two VLCPUFAs in human cells. Compared to docosahexaenoic acid (DHA), the golden standard in evaluating the health-promoting activities of VLCPUFAs, both DDA and DTA exhibited comparable or even better antitumor and antioxidant effects against human breast cancer SK-BR-3 and MDA-MB-231 cells. Especially, DTA elicited much stronger antioxidant and pro-apoptotic effects than DHA. Furthermore, DDA and DTA showed strong anti-inflammatory effects in human macrophages differentiated from monocyte THP-1 cells through lowering the protein expression levels of pro-inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), interferon γ (IFN-γ), monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor α (TNF-α). Future In Vivo and In Vivo studies are warranted to identify the mechanism of action (MOA) for the antitumor, antioxidant and anti-inflammatory functions of DDA and DTA and explore potential applications of these two VLCPUFAs as novel nutraceutical supplements in preventing inflammatory conditions, aging and even cancer.