?3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2Htetrazolium (MTS) assay5 × 103
/wells MHCC97H and Hep3B cells were seeded in a 96-well plate. Cells were transfected with indicated plasmids after being cultured for 24 hours. 20 μL MTS buffer (K300-500, Amyjet Scientificlnc, China) was added to each well and incubated for 2-4 hours after cells were transfected for 24 hours. The absorbance value was measured at 450 nm. Five duplicated wells were set for each group. Cell survival rates = (optical density (OD)EXP ? ODEmpty/ODControl ? ODEmpty)%.The experiment was repeated 3 times.

Transwell assay
Cells were resuspended using the DMEM supplemented with 10 g/L bovine serum albumin (BSA) and adjusted to the final density of 1 × 104 cells/mL. The Transwell chamber was put into a 24-well plate, and the upper chamber surface of the bottom membrane of the Transwell chamber was coated with Matrigel (40111ES08, Yeasen BioTechnologies co., Ltd., Shanghai, China) at the ratio of 1:8 and air-dried overnight at 4°C. After routine digestion, cellswere resuspended in a culture medium, adjusted to a cell density of 
1 × 105 cells/mL, and 200 μL of the cell suspension was added to the top of the Transwell chamber covered with Matrigel (BD, San Jose, CA, USA). The lower chambers were supplemented with 600 μL medium containing 20% FBS. The Transwell chamber was collected following 24 hours of incubation at 37°C, where the Matrigel and cells were removed with a cotton swab, and subjected to fixation and then stained by crystal violet dissolved in methanol. Stained cells were counted in five random fields per well using an inverted 
microscope (XDS-800D; Caikon, Shanghai, China). The invasive cells were counted. Triplicates were set in each group. This experiment was repeated for at least 3 times.

感謝中國上海市同濟大學醫學院附屬第十人民醫院醫學研究中央實驗室引用文獻