HERG suppresses the malignant phenotypes of osteosarcoma via modulating NF-κB pathway

doi:10.3760/cma.j.issn.1673422X.2016.07.007

Abstract

Abstract: ObjectiveTo detect the expression of HERG (human etheràgogorelated gene) potassium channel in human osteosarcoma, and explore the effects of silencing HERG by small interfering RNA (siRNA) on the proliferation and apoptosis of osteosarcoma cells and the mechanisms responsible for HERG regulation. MethodsThe expressions of HERG in osteosarcoma MG63 cells and tissues were detected by reverse transcription polymerase chain reaction (RTPCR), Western blotting and immunohistochemistry. Next, osteosarcoma cells were divided into three groups: HERGsiRNA group, controlsiRNA group and blank group. CCK8, colony formation, flow cytometry and Tunel assay were used to measure the proliferation and apoptosis of the osteosarcoma cells. Finally, Western blotting analysis was performed to detect the expression of nuclear factorκB (NFκB) pathway in osteosarcoma cells treated with HERG siRNA. ResultsOsteosarcoma cells and tissues were found to highly express HERG. Inhibition of HERG in the osteosarcoma cells significantly inhibited the cell proliferation and induced cell apoptosis. Compared to controlsiRNA group or blank group, HERGsiRNA could inhibit the proliferation of MG63 cells significantly ［HERGsiRNA group: (75.34±4.45)%; compared to controlsiRNA group: (100.60±5.31)%; t=3.64, P=0.007; compared to blank group: (100.00±5.66)%; t=3.43, P=0.009］. The similar results were obtained from colony formation assay (HERGsiRNA group: 134.30±11.82; compared to controlsiRNA group: 225.30±11.56; t=5.51, P=0.002; compared to blank group: 232.80±12.21; t=5.80, P=0.001). HERGsiRNA transfected MG63 cells demonstrated a significant increase of apoptotic rate compared to controlsiRNA transfected cells or untreated cells ［HERGsiRNA group: (28.10±2.21)%; compared to controlsiRNA group: (9.36±2.42)%; t=5.72, P=0.005; compared to blank group: (10.92±2.51)%; t=5.14, P=0.007］. This result was further confirmed by Tunel assay. The cells transfected with HERGsiRNA (31.57±2.08)% demonstrated extensive apoptosis, compared with the controlsiRNA group ［(10.35±1.82)%; t=7.69, P=0.002)］ or blank group ［(7.96±0.88)%; t=10.48, P=0.001］. Silencing HERG gene downregulated the cIAP1, XIAP, Bcl2, Survivin, PIκBα and NFκB p65 expression, compared to the control groups. ConclusionHERG is highly expressed in osteosarcoma. HERG silencing can suppress osteosarcoma progression through NFκB pathway and suggest that HERG may be a novel molecular target for osteosarcoma therapy and diagnosis.

Key words: Potassium channels, Osteosarcoma, Cell proliferation, Apoptosis, , NF-kappa B

WU Jin, CHEN Zhi-Da, ZENG Wen-Rong, LIN Bin, WU Xin-Yu, LIU Qing-Jun. HERG suppresses the malignant phenotypes of osteosarcoma via modulating NF-κB pathway[J]. Journal of International Oncology, 2016, 43(7): 508-514.

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