title: TALENs-directed knockout of the full-length transcription factor Nrf1α that represses malignant behaviour of human hepatocellular carcinoma (HepG2) cells.
abstract: The full-length Nrf1α is processed into distinct isoforms, which together regulate genes essential for maintaining cellular homeostasis and organ integrity, and liver-specific loss of Nrf1 in mice results in spontaneous hepatoma. Herein, we report that the human constitutive Nrf1α, rather than smaller Nrf1β/γ, expression is attenuated or abolished in the case of low-differentiated high-metastatic hepatocellular carcinomas. Therefore, Nrf1α is of importance in the physio-pathological origin and development, but its specific pathobiological function(s) remains elusive. To address this, TALENs-directed knockout of Nrf1α, but not Nrf1β/γ, is created in the human hepatocellular carcinoma (HepG2) cells. The resulting Nrf1α(-/-) cells are elongated, with slender spindle-shapes and enlarged gaps between cells observed under scanning electron microscope. When compared with wild-type controls, the invasive and migratory abilities of Nrf1α(-/-) cells are increased significantly, along with the cell-cycle G2-M arrest and S-phase reduction, as accompanied by suppressed apoptosis. Despite a modest increase in the soft-agar colony formation of Nrf1α(-/-) cells, its loss-of-function markedly promotes malgrowth of the subcutaneous carcinoma xenograft in nude mice with hepatic metastasis. Together with molecular expression results, we thus suppose requirement of Nrf1α (and major derivates) for gene regulatory mechanisms repressing cancer cell process (e.g. EMT) and malignant behaviour (e.g. migration).
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Ren
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Qiu
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Fenglin
authors.initials: F
authors.lastname: Lü
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Xufang
authors.initials: X
authors.lastname: Ru
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Shaojun
authors.initials: S
authors.lastname: Li
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Yuancai
authors.initials: Y
authors.lastname: Xiang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, and Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University Health Science Center, No 38 Xueyuan Rd., Haidian District, Beijing 100191, China.
authors.forename: Siwang
authors.initials: S
authors.lastname: Yu
authors.affiliation: The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China.
authors.forename: Yiguo
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002465:Cell Movement; D002471:Cell Transformation, Neoplastic; D004195:Disease Models, Animal; D055786:Gene Knockout Techniques; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D064593:Heterografts; D006801:Humans; D008819:Mice, Nude; D008855:Microscopy, Electron, Scanning; D051266:Nuclear Respiratory Factor 1 / Q000235:genetics*; D012878:Skin Neoplasms / Q000473:pathology; D000069896:Transcription Activator-Like Effector Nucleases / Q000378:metabolism*; D014410:Tumor Stem Cell Assay
chemical_list: C495633:NRF1 protein, human; D051266:Nuclear Respiratory Factor 1; D000069896:Transcription Activator-Like Effector Nucleases