Abstract
Objective To explore the effects of extracellular signal-regulated kinase1 (ERK1) small interfering RNA on the biological characteristics of rat hepatoma cells and its possible mechanism.Methods The expressions of ERK1 in hepatoma cell lines,primary rat hepatocyte,and 12 human hepatocellular carcinoma (HCC) specimens and their paired non-cancerous liver tissues were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry staining.Hepatoma cell lines RH-35 and H4IIE cells were infected with extracellular signal-regulated kinase 1 by adenvirus mediated small interfering RNA (AdshERK1) or a scrambled small interfering RNA used as a negative control which was mediated by adenvirus (AdshNC).After the expression of ERK1 in hepatoma cells downregulated,the effects on its biological characteristics and the changes of its downstream oncogene expression were detected.The data were analyzed by t test.Results Among 12 human HCC specimens,the expressions of ERK1 in 11 specimens were higher than those of corresponding adjacent tissues.Compared with primary hepatocytes,the expressions of ERK1 in rat hepatoma cell lines RH 35 and H4IIE upregulated (0.016±0.003 vs 0.075±0.010,0.458±0.052;t=13.806 and 7.715,both P＜0.01).After RH-35 and H4IIE cells transfected with AdshERK1,the ability of cell proliferation and colony formation were suppressed.Compared with AdshNC,AdshERK1 transfection could significantly decrease the percentage of S phase cells(18.55％ ±4.80％ vs 4.39 ％ ± 0.85 ％,t =-6.826,P＜0.01),which caused G0/G1 phase of cell cycle arrest(76.77 ％ ±0.93％ vs 89.57％ ± 1.26％,t =11.676,P＜0.01) ; AdshERK1 significantly inhibited the tumor formation ability of H4IIE cells in nude mice [(0.603± 0.284) mg vs (0.051 ± 0.074) mg,t=-5.340,P＜ 0.01].AdshERK1 downregulated the expressions of ERK1 in RH-35 and H4IIE cells and suppressed the expressions of downstream oncogene c-myc.Conclusions AdshERK1 has the role of inhibiting cell proliferation,colony and tumor formation ability of RH-35 and H4IIE cells.This role may be closely related with inhibiting the expression of its downstream oncogene c-myc. Key words: Carcinoma, hepatocellular; Mitogen-activated protein kinase 3; RNA, small interfering; Genes, myc; Tumor cells, cultured
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