Antimelanoma activity of perphenazine and prochlorperazine in human COLO829 and C32 cell lines

Abstract

Cutaneous melanoma is least common (only about 1% of skin cancers) but is the deadliest malignant tumor. Moreover, amelanotic types of melanoma are very difficult for clinical diagnosis. The standard therapy can cause a lot of side effects, e.g., nausea, vomiting, and headaches, which means that novel and effective strategies are required. Interestingly, phenothiazine derivatives possess sedative, antiemetic, and anticancer activity. Our goal was to determine the effect of perphenazine and prochlorperazine on cell viability, motility, microphthalmia-associated transcription factor (MITF) and tyrosinase content in melanotic and amelanotic melanoma cells. The viability of C32 and COLO829 melanoma cells was evaluated by the WST-1 colorimetric assay; impact on motility of human melanoma was performed by wound-healing assay, while tyrosinase and MITF content were determined by Western blot. In the present study, we explore the anticancer effect of perphenazine and prochlorperazine in human melanotic (COLO829) and amelanotic (C32) melanoma cells concluding that prochlorperazine inhibits cell viability in a concentration-dependent manner, impairs motility, and decreases tyrosinase and MITF amounts. Moreover, the analyzed drugs decrease/increase MITF amount depending on the type of melanoma. We demonstrated that the decrease of MITF and tyrosinase protein induces motility inhibition of C32 cells, which suggests the ability of those drugs to restore cancer cell sensitivity to treatment. The ability of prochlorperazine to contain the spread of the amelanotic melanoma in vivo may be helpful in the development of a new and effective antimelanoma therapies.

In vitro anticancer activity of fluphenazine, perphenazine and prochlorperazine. A review

“…Interestingly prochlorperazine at a concentration of 3 μ m more efficiently reduced motility than perphenazine. Moreover, the analysis of MITF and tyrosinase levels showed an increase in pigmented COLO829 melanoma cells and a decrease in C32 cells, which suggests the role of melanin pigment in amelanotic melanoma therapy as amelanotic cells are more sensitive to perphenazine and prochlorperazine treatment than melanotic cells (Otręba et al, 2019).…”

“…In addition, as the significant differences in comparison with the control were observed after 6, 9, 12 and 24 hours for the same cell line, it suggests that prochlorperazine could contain the spread of amelanotic melanoma in vivo. The analysis of microphthalmia-associated transcription factor (MITF) levels, crucial growth and survival factor, in both cell lines, showed a dose-dependent increase of MITF levels in COLO829 cells and a significant decrease of MITF level only after prochlorperazine treatment in C32 cells (Otręba et al, 2019…”

“…Otręba, Pajor and Warncke (2019) measured antimelanoma activity of perphenazine and prochlorperazine towards human melanotic (COLO829) and amelanotic (C32) cell lines. The EC 50 values calculated based on viability results were 3.99 and 2.62 μ m for perphenazine as well as 3.76 and 2.90 μ m for prochlorperazine, respectively for COLO829 and C32 cells.…”

“…In addition, as the significant differences in comparison with the control were observed after 6, 9, 12 and 24 hours for the same cell line, it suggests that prochlorperazine could contain the spread of amelanotic melanoma in vivo. The analysis of microphthalmia‐associated transcription factor (MITF) levels, crucial growth and survival factor, in both cell lines, showed a dose‐dependent increase of MITF levels in COLO829 cells and a significant decrease of MITF level only after prochlorperazine treatment in C32 cells (Otręba et al, 2019). Previously, measured EC 50 values for normal human epidermal melanocytes (HEMn) that were darkly and lightly pigmented were 2.76 (Otręba, Wrześniok, Beberok, Rok, & Buszman, 2016) and 18.49 μ m (Otręba, Wrześniok, Rok, Beberok, & Buszman, 2017), respectively for perphenazine and prochlorperazine towards HEMn darkly pigmented cells, as well as 0.63 μ m (Otręba, Beberok, Wrześniok, & Buszman, 2018) for prochlorperazine‐treated HEMn lightly pigmented cells.…”

“…Interestingly prochlorperazine at a concentration of 3 μM more efficiently reduced motility than perphenazine. Moreover, the analysis of MITF and tyrosinase levels showed an increase in pigmented COLO829 melanoma cells and a decrease in C32 cells, which suggests the role of melanin pigment in amelanotic melanoma therapy as amelanotic cells are more sensitive to perphenazine and prochlorperazine treatment than melanotic cells(Otręba et al, 2019).A summary of the anticancer effect (excluding the impact on cellular viability) of FLU, perphenazine and prochlorperazine towards different human cancer cell lines is presented in Figure 1. Based on the publications analyzed in the literature it is not possible to show the precise mechanistic pathway of the analyzed drugs because more accurate experiments are needed.…”