Tetracycline

L'espressione di CK2 indotta da Tetracycline aumenta significativamente l'invasione del carcinoma polmonare non a piccole cellule (NSCLC) in cellule che esprimono BRMS1 wild-type, ma non la mutante S30A.[2]

• Linee cellulari

  H157 BRMS1 knockdown (KD) cells, H157 stable cells

• Concentrazioni

  1 µg/mL

• Tempo di incubazione

  48 h

• Metodo

  H157 BRMS1 knockdown (KD) cells were cotransfected with pcDNA3-luciferase and Tet-on CK2α'. Stable clones were selected by Geneticin (400 µg/mL), Blasticidin (5 µg/mL), and Zeocin (150 µg/mL). Myc/His(6)- CK2α' expression was confirmed by immunoblot after treatment with TCN (1 µg/mL) for 48 h. Then, the H157 BRMS1^KD/luciferase/Tet-on CK2α' cells were infected with pBabe-shRNA–resistant FLAG-BRMS1 wild-type, S30A mutant, or empty vector and selected with puromycin (1 µg/mL). Flag-BRMS1 expression was confirmed by Western blot. H157 stable cells were pretreated with or without TCN (1 µg/mL) for 48 h. the inserts were coated with type IV collagen at a concentration of 0.25 mg/ml. The bottom portion of the wells included 1% FBS-media. Cells were allowed to migrate for 6 hours or invade for 22 hours. At the specified time point, inserts were fixed and stained with crystal violet.