Invasion Assay for Breast Cancer

Boyden Chamber assay was performed to access invasion capability of breast cancer cells according to earlier reported method with slight modifications.^{15 (link)} Breast cancer cells were treated with CoCl2 (100 µM)) and transfected with IL-6R siRNA. Other groups were treated with Dia (6 µM) or 5-Aza (5 µM). These treated cells were placed on the upper chamber of Boyden set up. In lower chamber, VEGF containing medium was added. After 24 h of incubation, upper chamber was taken and non-migrated cells were swapped with cotton plug. Upper chambers were then fixed with methanol and stained with haematoxylin and eosin followed by proper washing. Migrated cells were subjected to bright field microscopy at ×20 magnifications (Carl Zeiss).