Guohua Zhu1, Haiping Dai2, Yuanxun Duan1, Zelin Yu1. 1. First Clinical College, Nanjing University of Chinese Medicine, Nanjing 210023, China. 2. Department of Hematology, First Hospital Affiliated to Suzhou University, Suzhou 215006, China.
Abstract
OBJECTIVE: To investigate the effect of silencing LPXN expression by RNA interference on the proliferation and drug sensitivity of human acute monocytic leukemia SHI-1 cells in vitro. METHODS: Small interfering RNA (siRNA) sequences targeting LPXN were designed and transiently transfected in SHI-1 cells via Lipofectamine 2000, and the most efficient siRNA sequence for LPXN silencing was identified using Western blotting. The protein expression levels of LPXN, p-JNK, p-P38 MAPK and p-ERK were in the cells transfected with the selected siRNA were detected using Western blotting, and the cell proliferation changes were assessed using CCK-8 reagent. RESULTS: LPXN silencing by siRNA transfection resulted in significant proliferation suppression in SHI-1 cells with an inhibition rate of(27.04±2.05) % (P < 0.05). Western blotting showed that treatment of the siRNA-transfected SHI-1 cells with 0-25 μmol/L curcumin or with 0-2.0 μmol/L Ara-C further increased the cell inhibition rate and obviously enhanced the expressions of p-P38 MAPK and p-JNK without significantly affecting p-ERK expression. CONCLUSIONS: Down-regulation of LPXN expression by siRNA transfection can suppress the proliferation and increase the drug sensitivity of SHI-1 cells probably by activating JNK and P38 MAPK.
OBJECTIVE: To investigate the effect of silencing LPXN expression by RNA interference on the proliferation and drug sensitivity of human acute monocytic leukemiaSHI-1 cells in vitro. METHODS: Small interfering RNA (siRNA) sequences targeting LPXN were designed and transiently transfected in SHI-1 cells via Lipofectamine 2000, and the most efficient siRNA sequence for LPXN silencing was identified using Western blotting. The protein expression levels of LPXN, p-JNK, p-P38 MAPK and p-ERK were in the cells transfected with the selected siRNA were detected using Western blotting, and the cell proliferation changes were assessed using CCK-8 reagent. RESULTS:LPXN silencing by siRNA transfection resulted in significant proliferation suppression in SHI-1 cells with an inhibition rate of(27.04±2.05) % (P < 0.05). Western blotting showed that treatment of the siRNA-transfected SHI-1 cells with 0-25 μmol/L curcumin or with 0-2.0 μmol/L Ara-C further increased the cell inhibition rate and obviously enhanced the expressions of p-P38 MAPK and p-JNK without significantly affecting p-ERK expression. CONCLUSIONS: Down-regulation of LPXN expression by siRNA transfection can suppress the proliferation and increase the drug sensitivity of SHI-1 cells probably by activating JNK and P38 MAPK.
Entities:
Keywords:
SHI-1 cells; acute monocytic leukemia; drug sensitivity; leupaxin; small interfere RNA
Authors: Silke Kaulfuss; Michal Grzmil; Bernhard Hemmerlein; Paul Thelen; Stefan Schweyer; Jürgen Neesen; Lukas Bubendorf; Andrew G Glass; Hubertus Jarry; Bernd Auber; Peter Burfeind Journal: Mol Endocrinol Date: 2008-05-01
Authors: Murugendra S Vanarotti; David B Finkelstein; Cristina D Guibao; Amanda Nourse; Darcie J Miller; Jie J Zheng Journal: Biochemistry Date: 2016-02-24 Impact factor: 3.162