Knockdown and Overexpression of HCC Regulators

Human HCC cell lines (PLC-8024, Huh7, Hep3B, MHCC-97H and HepG2) and MIHA normal liver cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China) with short tandem repeat appraisal certificates. All cells were cultured as described previously.21 (link) Huh7 and MHCC-97H cells were infected with lentiviral pLKO.1 particles containing short hairpin RNA (shRNA) and were selected with 3 mg/mL puromycin (ThermoFisher, Waltham, MA, USA) for 4 days. Lentiviral pLKO.1 plasmids for shPLOD2 (Jidan, Guangzhou, China) were packaged with Lenti-Pac HIV mix (GeneCopoeia, Guangzhou, China) and Endo-Fectin Lenti (GeneCopoeia, Guangzhou, China) in 293T cells to produce lentiviral particles. For BIRC3 overexpression experiment, the BIRC3 expression plasmid (Jidan, Guangzhou, China) was packaged with Lipofectamine 2000 (ThermoFisher, Waltham, MA, USA) and then infected PLOD2 knockdown (KD) Huh7 and MHCC-97H cells.

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Li K., Niu Y., Li K., Zhong C., Qiu Z., Yuan Y., Shi Y., Lin Z., Huang Z., Zuo D., Yuan Y, & Li B. (2023). Dysregulation of PLOD2 Promotes Tumor Metastasis and Invasion in Hepatocellular Carcinoma. Journal of Clinical and Translational Hepatology, 11(5), 1094-1105.

Publication 2023

puromycin Lenti-Pac HIV mix Endo-Fectin Lenti Lipofectamine 2000

293t cells Birc3 Cell lines Cells Chinese Endo Human Lipofectamine 2000 Liver Miha Plasmid Puromycin Short tandem repeat Shrna

Corresponding Organization :

Other organizations : Sun Yat-sen University Cancer Center, Sun Yat-sen University

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Variable analysis

independent variables

Lentiviral pLKO.1 particles containing short hairpin RNA (shRNA) for PLOD2 knockdown
BIRC3 expression plasmid for overexpression

dependent variables

Not explicitly mentioned

control variables

Human HCC cell lines (PLC-8024, Huh7, Hep3B, MHCC-97H and HepG2)
MIHA normal liver cells
Culture conditions as described previously

controls

Positive control: Not specified
Negative control: Not specified

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