Effect of cadmium on the expression levels of interleukin-1a and interleukin-10 cytokines in human lung cells

ODEWUMI, CAROLINE; LATINWO, LEKAN M.; SINCLAIR, ANDRE; BADISA, VEERA L.D.; ABDULLAH, AHKINYALA; BADISA, RAMESH B.

Preparation of cell extracts. A total of ~3.9x10 6 A549 lung cancer cells or 1.95x10 6 normal MRC-9 lung cells were plated into T-75 cm 2 flasks in complete medium. Each cell line was treated with 0, 50, 100 or 150 µM CdCl 2 in triplicate flasks for various durations (0, 6, 12, 18 or 24 h) and were incubated at 37˚C at 5% CO 2 . At the end of each treatment period, the cells were trypsinized and centrifuged at 1,350 x g for 5 min. The cells were lysed in 1 ml of 50 mM potassium phosphate (pH 7.0) lysis buffer, containing 0.1% Triton X-100, by homogenization in a vial on ice three times for 10 sec using a polytron homogenizer (Pro Scientific, Inc., Oxford, CT, USA). The homogenate was subsequently transferred to an eppendorf tube and centrifuged at 12,740 x g for 10 min at 4˚C to remove the lysed cell membrane debris. The supernatants were transferred into fresh tubes and the cell lysates were stored at -20˚C for cytokine analysis.