To optimize the effective dose of Mitomycin C (MMC), 5 fluorouracil (5FU), and combination of MMC and 5FU on basal cell carcinoma (BCC)-derived cultivated cells and making a comparison between the treated and control groups.

Fresh specimens of BCC cases were cultivated and immune characterized via immunocytochemistry for BER-EP4 and EMA. Then BCC cells and fibroblastic cells were treated with different concentrations of MMC, 5FU, and combination of MMC and 5FU. In addition to MTT assay, flowcytometry for investigating cell cycle and apoptosis, and RT-PCR for TP53, CDKN1A, and CDK6 gene expression were performed.

BCC cells were immunoreactive for BER-EP4 but not for EMA. Based on MTT results, the concentration of 0.312 mg/ml was considered optimum for both MMC and 5FU. Mean viability of treated BCC cells with combinaton of MMC and 5FU was not significantly different from controls; however, means of those treated with MMC alone and 5FU alone were significantly less than controls (P = 0.002 and P = 0.04, respectively). Flowcytometry showed higher apoptotic rates in BCC cells treated with MMC (P = 0.002) and cell cycle arrest in phase S in all treated BCC groups. Gene expression of TP53 was significantly increased in MMC-treated BCC cells (P = 0.0001). CDKN1A gene was upregulated and CDK6 gene was downregulated in BCC cells treated with either 5FU (P = 0.0001 and P = 0.01, respectively) or combination of MMC and 5FU (P = 0.007 and P = 0.001, correspondingly).

The optimum dose of MMC induces more apoptosis in treated BCC cells as compared with 5FU and the combination of MMC and 5FU, while inducing a limited effect on fibroblasts apoptosis. Our results can be promising for non-surgical treatment of eyelid BCC with minimum side effects on adjacent healthy cells.