Figure 6From: Homozygous deletion of the activin A receptor, type IB gene is associated with an aggressive cancer phenotype in pancreatic cancerRole of ACVR1B gene in colony formation and cellular growth. To evaluate the role of the ACVR1B gene, we used stable ACVR1B-knockdown cell lines (shACVR1B) or control (shScr). (A) Western blot analyses after activin A stimulation (10 ng/mL). In contrast to Sui66/shScr and Sui73/shScr cell lines, ACVR1B was not expressed in Sui66/shACVR1B and Sui73/shACVR1B cell lines. SMAD2 was not phosphorylated by activin A in Sui66/shACVR1B and Sui73/shACVR1B cell lines. β-actin was used as an internal control. (B) Influence of activin A on cellular growth. The cell lines were stimulated with or without activin A (10 ng/mL) for 72 hours. Cell proliferation was assayed using an MTT assay. Although activin A inhibited cellular growth in Sui66/shScr and Sui73/shScr cell lines (Sui66/shScr-1; P = 0.0014*, Sui66/shScr-2; P = 0.0032*, Sui73/shScr-1; P = 0.0085*, and Sui73/shScr-2, P = 0.0053*, respectively), it did not influence the cellular growth in Sui66/shACVR1B and Sui73/shACVR1B cell lines (Sui66/shACVR1B-1; P = 0.81, Sui66/shACVR1B-2; P = 0.73, Sui73/shACVR1B-1; P = 0.90, and Sui73/shACVR1B-2; P = 0.87, respectively). (C) Colony formation of Sui66- transfectant cell lines. The colony formations in Sui66/shACVR1B cell lines were enhanced, compared with that in the control cell lines (Scr-1, 46.11 ± 8.00 vs. ACVR1B-1, 83.00 ± 16.75, P = 0.026* and Scr-2, 52.55 ± 9.24 vs. ACVR1B-2, 103.70 ± 21.00, P = 0.017*). (D) Cellular growth of Sui73- transfectant cell lines. The cellular growth was evaluated using an MTT assay. The cellular growths in Sui73/shACVR1B cell lines were enhanced, compared with Sui73/shScr (shScr-1 vs. shACVR1B-1, 0 h, P = 081; 24 h, P = 0.88; 48 h, P = 0.063; 72 h, P = 0.013*, and shScr-2 vs. shACVR1B-2, 0 h, P = 040; 24 h, P = 0.45; 48 h, P = 0.040*; 72 h, P = 0.014*). Columns, mean of independent triplicate experiments; Lines, mean of independent triplicate experiments; Bars, SD; *P < 0.05.Back to article page