HEK 293 cell-line had the highest growth rate amongst the three cell lines. For TB assay, growth of HEK 293>THP-1 and for MTT, HEK 293>MCF-7 while the DT was in the order of THP-1>MCF-7>HEK 293. Sophorolipid showed anti-proliferative activity comparable to doxorubicin on THP-1>MCF-7>HEK 293. THP-1 showed high sensitivity to sophorolipid with IC50 of 10.50, 25.58 and 6.78(μg/ml) after 24, 48 and 72h respectively. However, sophorolipid was cytotoxic from 24 to 72h on HEK 293 cell lines with IC50 of 21.53, 40.57 and 27.53μg/ml respectively. Although, doxorubicin showed higher anti-proliferative activity than all biosurfactants, it had poorer selectivity index for the same time durations compared to the biosurfactants. This indicates that biosurfactants were more effective for slowing the growth of the tested cancer cell lines and hence may be potential candidates for use in human cancer therapy. Physico-chemical characteristics of the biosurfactants suggest that their mechanism of action may be due to activity on the cell membrane.