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- 2019
Large tumor suppressor kinase 2 overexpression attenuates 5-FU-resistance in colorectal cancer via activating the JNK-MIEF1-mitochondrial division pathwayDOI: 10.1186/s12935-019-0812-3 Keywords: 5-FU, Colorectal cancer, LATS2, Mitochondrial fission, MIEF1, Apoptosis Abstract: 5-FU treatment attenuates the levels of LATS2 and overexpression of LATS2 enhances 5-FU-mediated cancer cell death in vitro. a–b Different doses of 5-FU were added into the medium of SW480 cells and then proteins were isolated from cells. Western blotting was used to observe the alterations of LATS2 in response to 5-FU treatment. c RNA was isolated from SW480 cells and then the transcription of LATS2 mRNA was evaluated via qPCR. 5-FU treatment dose-dependently reduced the transcription of LATS2 in SW480 cells. d Cellular viability of SW480 cells was determined via MTT assay. e, f LATS2 adenovirus vectors (ad-LATS2) were transfected into SW480 cells and then the overexpression efficiency of LATS2 was confirmed via western blotting. g After LATS2 was overexpressed, the cell death was evaluated via LDH release assay. LATS2 overexpression could further elevate the LDH content in the medium. h Caspase-3 activity was evaluated via ELISA. LATS2 adenovirus vectors (ad-LATS2) were transfected into SW480 cells. i, j TUNEL staining was used to label the apoptotic cells. The number of TUNEL-positive cell was recorded to reflect the cell death in response to LATS2 overexpression. *p?<?0.05 vs. control (ctrl) group; #p?<?0.05 vs. 5-FU?+?ad-ctrl group; ad-ctrl: control null adenovirus vectors; ad-LATS2: LATS2 adenovirus vector
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