title: mRNA Expression of Bax, Bcl-2, p53, Cathepsin B, Caspase-3 and Caspase-9 in the HepG2 Cell Line Following Induction by a Novel Monoclonal Ab Hep88 mAb: Cross-Talk for Paraptosis and Apoptosis.
abstract: Monoclonal antibodies with specific antigens have been widely used as targeted therapy for cancer. Hep88 mAb is a monoclonal antibody which shows specific binding with anti-cancer effects against the HepG2 cell line. However, its mechanisms of action are still not completely understood. We examined cell cycling and apoptosis by flow cytometry and mRNA expression of factors involved in apoptosis and paraptosis in Hep88 mAb-treated HepG2 cells by real-time PCR. The cell-cycle analysis demonstrated that growth-inhibitory activity was associated with G2/M cell cycle arrest. Hep88 mAb induced a significant increase in apoptotic cell populations in a dose- and time-dependent manner. The mRNA expression results also suggested that the process triggered by Hep88 mAb involved up-regulation of tumor suppressor p53, pro-apoptotic Bax, Cathepsin B, Caspase-3 and Caspase-9, with a decrease of anti-apoptotic Bcl-2 - thus confirming paraptosis and apoptosis programmed cell death. These findings represent new insights into the molecular mechanisms underlying the anti-cancer properties of Hep88 mAb in liver cancer cells.
authors.affiliation: Faculty of Medicine, Thammasat University (Rangsit Campus), Pathum Thani, Thailand E-mail : panadda_rojpibulstit@hotmail.com.
authors.forename: Thantip
authors.initials: T
authors.lastname: Mitupatum
authors.forename: Kalaya
authors.initials: K
authors.lastname: Aree
authors.forename: Suthathip
authors.initials: S
authors.lastname: Kittisenachai
authors.forename: Sittiruk
authors.initials: S
authors.lastname: Roytrakul
authors.forename: Songchan
authors.initials: S
authors.lastname: Puthong
authors.forename: Sasichai
authors.initials: S
authors.lastname: Kangsadalampai
authors.forename: Panadda
authors.initials: P
authors.lastname: Rojpibulstit
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D053148:Caspase 3 / Q000235:genetics*; D053453:Caspase 9 / Q000235:genetics*; D002401:Cathepsin B / Q000235:genetics*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D003429:Cross Reactions; D005434:Flow Cytometry; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D051028:bcl-2-Associated X Protein / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; C494826:BAX protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D053148:Caspase 3; D053453:Caspase 9; C533912:CTSB protein, human; D002401:Cathepsin B