Bhupal Bhetwal PhD
Medicine and Health Sciences
Introduction: Bitter melon extract (BME) is known to inhibit breast cancer cells (MCF-7) proliferation. The PKC and ROK play critical roles in cell division, migration, and survival. However, the roles of protein kinase-c (PKC) and rho kinase (ROK) inhibition on MCF-7 cells have not been established. Moreover, whether potential effects of BME’s effects on MCF-7 cells are mediated by PKC and ROK are unknown. Aims: We aimed to investigate if BME exerts cytotoxic effects on breast cancer cells (MCF-7 cells) and if PKC and ROCK mediate BME’s effects. We hypothesized that BME inhibits proliferation of MCF-7 cells by decreasing PKC activity and increasing ROK activity. Methods: Fresh bitter melons were purchased from an Asian grocery store and the extract (BME) was extracted, centrifuged, and filter sterilized. The MCF-7 cells were cultured in DMEM medium with different amounts of BME [0%, 0.5%, 1%, 2%, 5%, and 10% of BME (v/v)], and in the presence of absence of PKC inhibitor (GF109203x; 0.5µM) and ROK inhibitor (H-1152; 1 µM). After culturing cells for 6 days (for the dose-response study) and 2 days (for the inhibitor studies), pictures of cultures were taken, cell viability was determined using Trypan blue dye, and change in protein expression was determined using western blotting. Repeated t-test was used to determine statistical significance. Results & Discussion: BME dose-dependently inhibited viability of MCF-cells (N=3) and MYPT1 (Myosin phosphatase targeting subunit-1) expression (N=1, duplicate). The PKC inhibitor alone did not have significant effect on cell viability (N=4) but the ROK inhibitor decreased cell viability (N=5) and MYPT1 expression. In the future, we will be studying phosphorylation of ROK’s target proteins to examine how ROK mediates BME’s cytotoxic effects.
Copyright 2018 all authors
Choi, Heeyun and Bhetwal, Bhupal Ph.D, "Role of Protein Kinase-C and Rho Kinase in the Cytotoxic Effects of Bitter Melon Extract on Metastatic Breast Cancer Cells" (2018). MU-COM Research Day. 102.