title: Knockdown of Golgi phosphoprotein 2 inhibits hepatocellular carcinoma cell proliferation and motility.
abstract: Golgi phosphoprotein 2 (GP73) is highly expressed in hepatocellular carcinoma (HCC) cells, where it serves as a biomarker and indicator of disease progression. We used MTS assays, anchorage-independent cell colony formation assays and a xenograft tumor model to show that GP73-specific siRNAs inhibit HCC proliferation in HepG2, SMMC-7721, and Huh7 cell lines and in vivo. Following GP73 silencing, levels of p-Rb, a factor related to metastasis, were reduced, but cell cycle progression was unaffected. Our results suggest that GP73 silencing may not directly suppress proliferation, but may instead inhibit cell motility. Results from proliferation assays suggest GP73 reduces expression of epithelial mesenchymal transition (EMT)-related factors and promotes cell motility, while transwell migration and invasion assays indicated a possible role in metastasis. Immunofluorescence co-localization microscopy and immunoblotting showed that GP73 decreases expression of N-cadherin and E-cadherin, two key factors in EMT, which may in turn decrease intracellular adhesive forces and promote cell motility. This study confirmed that GP73 expression leads to increased expression of EMT-related proteins and that GP73 silencing reduces HCC cell migration in vitro. These findings suggest that GP73 silencing through siRNA delivery may provide a novel low-toxicity therapy for the inhibition of tumor proliferation and metastasis.
authors: Medical Biotechnology Laboratory, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors: Yiming
authors: Y
authors: Liu
authors: Medical Biotechnology Laboratory, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors: Xiaodi
authors: X
authors: Zhang
authors: Department of Pathology, Zhejiang Provincial People's Hospital, Hangzhou 310014, China.
authors: Ting
authors: T
authors: Sun
authors: Department of Pathology, RunRun-Shaw Hospital Affiliated to Zhejiang University School of Medicine, Hangzhou 310016, China.
authors: Junchang
authors: J
authors: Jiang
authors: Clinical Laboratory, Children's Hospital Affiliated to Zhejiang University School of Medicine, Hangzhou 310003, China.
authors: Ying
authors: Y
authors: Li
authors: Medical Biotechnology Laboratory, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors: Mingliang
authors: M
authors: Chen
authors: Laboratory Animal Center, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors: Zhen
authors: Z
authors: Wei
authors: Cancer Center, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
authors: Weiqin
authors: W
authors: Jiang
authors: Medical Biotechnology Laboratory, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors: Linfu
authors: L
authors: Zhou
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D015151:Immunoblotting; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D018613:Microscopy, Confocal; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014183:Transplantation, Heterologous
chemical_list: D015820:Cadherins; C410824:GOLM1 protein, human; D008565:Membrane Proteins
keywords: Golgi phosphoprotein 2; biomarker; cell proliferation; hepatocellular carcinoma; siRNA