Evaluation of the anticancer effects of kaempferol using in vitro assays and [99mTc]Tc-PSMA SPECT/CT imaging in a DMBA-induced breast cancer model

Background Kaempferol is a naturally occurring flavonoid with reported anticancer properties; however, its effects on breast cancer cells and tumor-associated molecular imaging require further investigation. This study evaluated the effects of kaempferol using complementary in vitro assays and a 7,12-dimethylbenz [a]anthracene (DMBA)-induced breast cancer rat model assessed by [ 99m Tc]Tc-PSMA SPECT/CT imaging. Methods MCF-7 breast cancer cells were treated with kaempferol, and cell viability was assessed using the MTT assay across a concentration range of 0–80 µM. Apoptosis, wound closure, and the expression of E-cadherin and VEGF were also evaluated. The half-maximal inhibitory concentration (IC 50 ) was estimated using nonlinear four-parameter logistic regression of replicate-level concentration–response data. In vivo , 8-week-old rats were allocated into cancer control, kaempferol pre-treatment, and post-treatment groups (n = 10/group). Mammary tumors were induced using DMBA, and animals received kaempferol (100 mg/kg/day, oral gavage) according to the experimental protocol. Tumor-associated tracer uptake was evaluated using [ 99m Tc]Tc-PSMA SPECT/CT imaging and quantified using tumor-to-background ratios (TBRs). Results Kaempferol reduced MCF-7 cell viability in a concentration-dependent manner, with nonlinear regression estimating an IC 50 of approximately 40 µM. Kaempferol treatment was associated with increased apoptosis, reduced wound closure, increased E-cadherin expression, and decreased VEGF expression. In vivo imaging demonstrated significant differences in tumor tracer uptake among the experimental groups. The pre-treatment group showed significantly lower TBR values (64.65 ± 1.69) than the cancer control (71.18 ± 1.83) and post-treatment groups (71.16 ± 1.06), whereas no significant difference was observed between the cancer control and post-treatment groups. One-way ANOVA demonstrated a significant overall group effect (F (2,27) = 58.08, p < 0.001, η 2 = 0.81). Conclusion Kaempferol demonstrated concentration-dependent cytotoxic effects and modulation of cancer-associated cellular markers in MCF-7 cells. In the DMBA-induced breast cancer model, reduced tumor-associated [ 99m Tc]Tc-PSMA uptake was observed predominantly following kaempferol pre-treatment, suggesting a potential chemopreventive effect. The findings support the complementary use of in vitro pharmacological assays and molecular imaging to investigate treatment-associated changes in tumor biology.