eradication of colorectal cancer stem cells through phyllanthus niruri linn extract induction: expression of tlr-4, nf-κb, and ctl in sprague-dawley rats treated with capecitabine chemotherapy

Chemoresistance in colorectal cancer (crc), driven by cancer stem cells (cscs) expressing cd133 and cd44, limits the efficacy of capecitabine through enhanced dna repair and immune evasion. phyllanthus niruri linn (pnl), known for its immunomodulatory and anticancer properties, may enhance treatment by targeting cscs and modulating immune pathways such as tlr-4, nf-κb, and cytotoxic t-lymphocytes (ctls). we hypothesized that pnl would suppress csc markers (cd133/cd44) and enhance ctl activity by inhibiting tlr-4/nf-κb signaling, thereby overcoming capecitabine resistance. a true experimental posttest-only control group design was conducted on 45 male sprague-dawley rats with 1,2-dimethylhydrazine (dmh)-induced colorectal cancer. after dropouts, 43 rats were analysed across five groups: normal control (kn), dmh-only (k-), capecitabine (k+), pnl (x1), and pnl+capecitabine (x2). the interventions were administered over 5 weeks following carcinogen induction. colonic tissue samples were collected post-euthanasia for immunohistochemical analysis of cd133, cd44, nf-κb, tlr-4, and ctl. semi-quantitative immunoreactivity scoring (irs) was used to assess marker expression. data were analysed using anova or kruskal-wallis tests. significant differences were observed in colon cecum weight ratios and body weight (p<0.001), cd133 (p=0.008), cd44 (p=0.004), and ctl (p=0.01) expression differed significantly across groups, with the lowest cd133/cd44 in the x2 group and the highest ctl in x1/x2. tlr-4 (p=0.212) and nf-κb (p=0.361) showed no significant changes. pnl, alone or combined with capecitabine, reduced csc markers and increased ctl infiltration, supporting its role in mitigating chemoresistance through immune modulation. while tlr-4/nf-κb pathways remained unaffected, pnl’s efficacy against cscs highlights its potential as an adjuvant therapy for crc.