title: TSG101 Silencing Suppresses Hepatocellular Carcinoma Cell Growth by Inducing Cell Cycle Arrest and Autophagic Cell Death.
abstract: BACKGROUND
  The tumor susceptibility gene 101 (TSG101) was originally identified as a tumor-suppressor gene that mediates many molecular and biological processes, such as ubiquitination, endosomal trafficking, cell survival, and virus budding, but its role in hepatocellular carcinoma (HCC) is currently unknown.
  
  
  METHODS
  We assessed the expression of TSG101 in HCC and paracancerous tissues using qPCR. Then, we used the TSG101-specific siRNA mix to disrupt the expression of TSG101 to investigate the subsequent effect on human hepatoma-7 (Huh7) cells. Western blot was used to detect the protein expression of TSG101 and other molecules. Cell growth assay was performed using CCK8. Transwell assay was used to investigate the migration and invasion ability of Huh7 cells after transfection with of TSG101 siRNA. Flow cytometry was used to estimate the effect of TSG101 knockdown on cell cycle and apoptosis. Confocal laser scanning microscopy was used to observe the actin filaments change and the formation of autophagy.
  
  
  RESULTS
  TSG101 was over-expressed in HCC tissues. TSG101 silence was able to suppress Huh7 cell proliferation, migration, and invasion. Furthermore, silencing of TSG101 could induce cell cycle arrest at G1 phase and inhibit the expression of cyclin A and cyclin D, while up-regulating the expression of CDK2. The mechanism might be induction of autophagic cell death and inactivation of Akt and ERK1/2.
  
  
  CONCLUSIONS
  TSG101 plays an important role in the development of HCC and may be a target for molecular therapy.
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Shao
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Weiping
authors.initials: W
authors.lastname: Ji
authors.affiliation: Department of General Surgery, Changzheng Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Anan
authors.initials: A
authors.lastname: Liu
authors.affiliation: Department of General Surgery, Suzhou Municipal Hospital, Suzhou, Jiangsu, China (mainland).
authors.forename: Ancheng
authors.initials: A
authors.lastname: Qin
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Li
authors.initials: L
authors.lastname: Shen
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Yingqi
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Xiangui
authors.initials: X
authors.lastname: Hu
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Enda
authors.initials: E
authors.lastname: Yu
authors.affiliation: Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
authors.forename: Gang
authors.initials: G
authors.lastname: Jin
mesh_terms: D017209:Apoptosis; D001343:Autophagy / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000628:therapy; D059447:Cell Cycle Checkpoints / Q000235:genetics*; D016923:Cell Death; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D019925:Cyclin A / Q000378:metabolism; D056741:Cyclin D / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics* / Q000502:physiology; D005434:Flow Cytometry; D016193:G1 Phase; D017353:Gene Deletion; D020868:Gene Silencing; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009361:Neoplasm Invasiveness; D034741:RNA, Small Interfering / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000502:physiology; D000099177:Tsg101 Protein
chemical_list: D019925:Cyclin A; D056741:Cyclin D; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D034741:RNA, Small Interfering; D014157:Transcription Factors; D000099177:Tsg101 Protein