Melanoma is one of the most aggressive types of cancer and extremely difficult to treat after metastasis. BRAFV600E-activating mutations give rise to ∼80% of melanocytic nevi, yet only one-third of melanocytic nevi result in melanoma, suggesting the involvement of other factors. Polo-like kinase 1 (PLK1), an important regulator of cell cycle progression, is overexpressed in melanoma and its expression has been shown to correlate with patient prognosis. Microphthalmia-associated transcription factor (MITF) is a melanocytic lineage-specific transcription factor that regulates a variety of genes critical for melanin synthesis as well as melanoma progression. The goal of this study was to define potential interactions between PLK1, BRAFV600E, and MITF in human melanoma. First, we employed a commercially available human tissue microarray (TMA) coupled with high-throughput, multispectral Vectra scanning and inForm analysis to study a number of clinical tissue cores (nevus, malignant and metastatic melanoma). The TMA was simultaneously immunostained for PLK1, BRAFV600E, MITF, proliferation marker Ki67, melanoma biomarker S100 and DAPI, and was subjected to Vectra scanning and inForm analyses. Using Simple Linear Regression analyses, we found significant correlations among each pair of the selected four proteins (PLK1, BRAFV600E, MITF and Ki67) with correlation co-efficient ranging 0.24-0.84. To analyze if PLK1, and BRAFV600E are contributing to cell proliferation (Ki67 expression) or affecting MITF expression, we employed a Multiple Linear Regression analysis. Our data suggested that high expressions of both BRAFV600E and PLK1 are correlated positively with the expression of Ki67. However, when fitting both PLK1 and BRAFV600E versus MITF, only high PLK1 had significant positive correlation with MITF, while BRAFV600E did not show correlation with MITF. These results suggest that PLK1 and MITF could contribute to melanoma progression independent to BRAFV600E. To further validate our findings, we analyzed The Cancer Genome Atlas (TCGA) database containing a large melanoma cohort of 432 melanoma patients with information on overall survival (OS). To visualize the survival plots, the expression level of PLK1 and MITF was sorted and equally separated to two groups using the median value as a cutoff. Kaplan-Meier analyses showed that high mRNA expression of both PLK1 and MITF were individually associated with significant reductions in OS. Interestingly, when we sorted the data for both high PLK1 and high MITF in the same patient, the OS was shorter than that of patients with low PLK1 and low MITF. Overall, our study suggests an association between PLK1 and MITF pathways during melanoma progression, which may affect overall survival in melanoma patients. Thus, concomitant targeting of PLK1 and MITF could provide an advantage over monotherapy towards melanoma management. However, in-depth studies are required to validate our findings. Citation Format: Gagan Chhabra, Shengqin Su, Chandra K. Singh, Mary A. Ndiaye, Nihal Ahmad. Potential correlations between PLK1, BRAF and MITF in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2157.