Qcm 24 well fluorimetric cell invasion assay kit

MDA-MB-231 and HCC-70 infected with Lv-NC, Lv-miR-890 or Lv-miR-890 and Lv-CD147 for 72 h were trypsinized and seeded into 96-well plates at a density of 1 × 10⁴ cells per well. The cells were cultured at 37 °C in 5% CO2. Cell viability was examined using CCK-8 (Cell Counting Kit-8, Dojindo, Japan) at 24, 48, and 72-h timepoints according to the kit instructions. Apoptosis was assessed using flow cytometry (FACS Calibur, BD, USA) with the Annexin V: FITC Apoptosis Detection Kit II (Cat:556570, BD) according to the instructions. Cell invasion experiments were performed using a QCM™ 24-well Fluorimetric Cell Invasion Assay Kit (Chemicon International, MI, USA) according to the manufacturer′s instructions. 500 μl medium supplemented with 10% FBS was used as a chemoattractant. 72 h after virus infection, 1 × 10⁵ cells of each group were seeded into the upper chamber and cultured in medium supplemented with 1% FBS for 12 h under 37 °C and 5% CO2. After 12 h, cells that invaded the underside of the membrane were fixed in 4% paraformaldehyde and stained with crystal violet staining solution, and the number of cells was counted by a fluorescence assay according to the kit instructions. The grouping was the same as that in the proliferation assay.

Cell invasion experiments were carried out with the QCM 24-well Fluorimetric Cell Invasion Assay kit (ECM554; Chemicon) according to the manufacturer’s instructions. Each insert contains an 8 μm pore size polycarbonate membrane coated with a thin layer of ECMatrixTM. The ECM layer occludes the membrane pores, blocking non-invasive cells from migrating through. Invasive cells, on the other hand, migrate through the ECM layer and cling to the bottom of the polycarbonate membrane. Invaded cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer and subsequently lysed and detected by CyQuant GR@ dye.