Caski

The human normal cervical cell lines (Hacat) and cervical cancer cell lines (HeLa, Siha, Caski, and C33a) were purchased from American Type Culture Collection (ATCC, USA). HeLa, Siha, C33a, and Hacat cells were incubated in DMEM medium (KeyGEN, Nanjing, China), and Caski cells were cultured in RPMI1640 (KeyGEN, Nanjing, China) medium containing 10% fetal bovine serum (GIBCO-BRL, Invitrogen, Carlsbad, CA, USA) and cultured at 37°C in a humidified incubator containing 5% CO₂.

Human cervical cancer cell lines SiHa, CaSki, Hela, Me180 and human non-tumor keratinocyte line HaCaT were obtained from Nanjing KeyGen Biotech Co, Ltd (Nanjing,China). The cells were cultured in Dulbecco’s modified Eagle’s medium (GIBCO, Carlsbad, California, USA) containing 10% FBS in a humidified atmosphere of 5% CO₂ at 37 °C. Human TFF3 expression, TFF3 siRNA and CDH1 siRNA plasmid constructs have been previously described [7 (link), 17 (link)]. Luciferase assays were performed as previously described [18 (link)]. Briefly, transfections were carried out in triplicate using 1 μg of the appropriate luciferase reporter construct and empty vector per transfection along with 0.1 μg of Renilla luciferase construct as control for transfection efficiency. Luciferase activities were assayed after 24 h of transfection using the Dual Luciferase Assay System (Promega Corp, Madison, WI, USA).