Cissus cornifolia (Baker) Planch is traditionally used in South African traditional medicine (SATM) to treat a variety of human infections. The antimicrobial activity of extracts from C. cornifolia was investigated in vitro against a plethora of pathogenic microorganisms using the microdilution assay. The acetone extract exhibited a notable minimum inhibitory concentration (MIC) value of 0.10 mg/mL against Mycoplasma hominis and a further MIC of 0.20 mg/mL against Candida parapsilosis, Streptococcus agalactiae, Pseudomonas aeruginosa, and Enterococcus faecalis. In the antiproliferative assays, both the ethyl acetate and methanol extracts exhibited a potent inhibition of the MCF-7-21 cell line. In the anti-inflammatory assays, both the ethyl acetate and methanol extracts exhibited IC50 values of 15.59 and 15.78 µg/mL against Cyclooxygenase-2 (COX-2), respectively. Methanol extract further exhibited potent dual inhibition of both COX-2 and 15-LOX enzymes, hence, recommended to curb both related cancers, particularly breast cancer and inflammation-borne diseases. In the comparative gas chromatography time-of-flight mass spectrometry (GC/TOF-MS), the acetone, ethyl acetate, and methanol extract contained significantly prevalent amounts of compound 2-(2’,4’,4’,6’,6’,8’,8’-Heptamethyltetrasiloxan-2’-yloxy)-2,4,4,6,6,8,8,10,10-nonamethylcyclopentasiloxane with % area ranging from 15.714 to 39.225. The findings in the current work validates the use of the plant species in SATM in the treatment of cancer-like infections, opportunistic infections associated with HIV-AIDS. Furthermore, the in vivo studies and the mechanisms of action still need to be explored.