Shilajit (Mumio) Elicits Apoptosis and Suppresses Cell Migration in Oral Cancer Cells through Targeting Urokinase-type Plasminogen Activator and Its Receptor and Chemokine Signaling Pathways

Background Shilajit a natural phytomineral has a proven record of treating many human body ailments. Objectives This study aimed to explore the influence of Shilajit on cell proliferation and apoptosis induction in oral cancer cells (OCC) and to comprehend the molecular mechanism associated with OCC migration. Materials and Methods Human gingival fibroblast (hGF) and OCC (KB-1 (KERATIN-forming tumor cell line HeLa)) were exposed to Shilajit solution dilutions. Cell growth and apoptosis were measured by MTT and Annexin-V tests (control/test group). Cellular morphology using an inverted microscope, cellular apoptosis using acridine orange/ethidium bromide dual staining, reactive oxygen species production analysis using 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, and gene protein expression using real-time polymerase chain reaction test were used to measure study outcomes. The data enumerated were the average of three trial experiments. Categorical and numerical data were expressed as frequency distribution and means, respectively. Differences in groups (control/test; zero/24 h/48 h) were determined using Student’s t -test and one way analysis of variance, with probability ‘ p ’ value considered significant at ≤0.05. Results The viability of OCC exhibited a concentration and time-dependent response to Shilajit. Notably, Shilajit demonstrated selectivity against cancer cells. Through an examination of the Annexin-V apoptosis assay, it was observed that Shilajit induces apoptosis by upregulating the proapoptotic gene expression ( p ≤ 0.05) and downregulating antiapoptotic proteins ( p ≤ 0.05). Furthermore, the impact of Shilajit on cell migration decreased significantly when compared to control cells through modulating the urokinase-type plasminogen activator (uPA) and its receptor (uPAR) and chemokines gene expression. Conclusion Shilajit exhibited greater cytotoxicity, decreased OCC proliferation and migration, and initiated OCC apoptosis as equivalent to normal cells. These promising outcomes indicate that Shilajit holds potential as a robust, promising option for oral cancer treatment, underscoring the need for further research in this domain.