Rescued and Recovered Ιn-Vitro Functions of Progesterone Treated Human Melanoma (BLM) Cells

Forty-eight hours of progesterone treatment of human melanoma (BLM) cells not only decreased cell growth (1), but also resulted in lot of floating cells. Addition of 2 mM of 3-methyl adenine (3-MA) partially rescued cell growth (2). Partial rescue in cell growth with 3-MA led us to check adhesion and migration functions as well because of floating cells. Partial rescue in cell growth also prompted us to check for recovery of those in-vitro functions after removing progesterone and 3-MA. Hence, the aim of the study was to compare in-vitro functions of progesterone treated, 3-MA rescued and recovered human melanoma cells. MTT proliferation assay, crystal violet adhesion assay and scratch migration assay were used to assess respectively in-vitro cell growth, adhesion and migration functions of melanoma cells.

Fifty μM of Progesterone treatment resulted in 42% cell growth compared to 100% growth in untreated control. But, 3-MA addition partially rescued cell growth to 46.7%. When both treated and rescued cells were allowed to recover initially for 48 hrs, P-50 μM treated cells showed improvement in cell growth to 50%, whereas 3-MA rescued cells showed 57.5% growth. Again, when both cells were allowed to recover for second 48 hrs, rescued cells showed a cell growth of 87%, whereas treated cells maintained at 52.9%, the previous level of growth. So rescued cells recovered cell growth better than treated cells. Partial recovery of cell growth in rescued cells prompted us to check adhesion function. P-50 μM treatment resulted in 71.8% adhesion compared to 100% in untreated control. But, 3-MA partially rescued adhesion to 76.9%. When both treated and rescued cells were allowed to recover for first 48 hrs, rescued cells showed improved adhesion (86.5%) than treated cells (71%). Again, when both cells were allowed to recover for second 48 hrs, rescued and treated cells showed almost equal recovery in adhesion 91.2% and 87.9% respectively. Following adhesion, migration function was checked. P-50 μM treatment decreased migration to 23% compared to 100% in untreated control. But 3-MA partially rescued migration to 45%. However, initial 48 hrs recovery did not improve migration function in both treated and rescued cells.

In conclusion, progesterone treatment decreased in-vitro cell growth, adhesion and migration functions of BLM cells. But 3-MA partially rescued cell growth, adhesion and migration functions. 3-MA rescued cells showed better recovery of in-vitro functions than progesterone treated cells.