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Many glioma patients develop resistance to temozolomide (TMZ) treatment, resulting in reduced efficacy and survival rates. TMZ‐resistant cell lines SHG44R and U87R, which highly express O6‐methylguanine DNA methyltransferase (MGMT) and P‐gp, were established. CN‐3, a new asterosaponin, showed cytotoxic effects on TMZ‐resistant cells in a dose‐ and time‐dependent manner via reactive oxygen species...