CAJ | 학술논문

目的探讨ERK1小干扰RNA对大鼠肝癌细胞株生物学特性的影响及其可能的作用机制.方法采用实时RT-PCR及免疫组织化学等方法检测ERK1在大鼠肝癌细胞株、大鼠原代肝细胞与12份人肝癌及配对癌旁组织中的表达,应用腺病毒介导的细胞外信号调节激酶1小干扰RNA(AdshERK1)感染大鼠肝癌细胞株RH-35、H4IIE,观察下调肝癌细胞中ERK1表达后对其生物学特性的影响及下游癌基因表达的变化.采用t检验进行统计学分析.结果 12份人肝癌组织中,有11份的ERK1表达较相应癌旁组织明显增多.ERK1在大鼠肝癌细胞株RH-35、H4IIE中的表达较大鼠原代肝细胞表达上调(0.075±0.010、0.458±0.052比0.016±0.003,t=13.806、7.715,P均＜0.01).AdshERK1感染RH-35、H4IIE细胞后,肝癌细胞增殖能力、克隆形成能力显著受抑制.与AdshNC组相比,AdshERK1可明显减少RH-35细胞中S期细胞比例(18.55％±4.80％比4.39％±0.85％,t=-6.826,P＜0.01),引起G0/G1期细胞周期阻滞(76.77％±0.93％比89.57％±1.26％,t=11.676,P＜0.01)；AdshERK1可显著抑制H4IIE细胞在裸鼠皮下成瘤能力[(0.603±0.284) mg比(0.051±0.074) mg,t=-5.340,P＜0.01].AdshERK1下调RH-35、H4IIE细胞ERK1表达,并抑制其下游癌基因c-myc的表达.结论 AdshERK1有抑制大鼠肝癌细胞株RH-35和H4IIE增殖、克隆形成及皮下成瘤能力的作用,且该作用可能与抑制ERK1下游肿瘤相关基因c-myc的表达密切相关.
목적 탐토ERK1소간우RNA대대서간암세포주생물학특성적영향급기가능적작용궤제.방법채용실시RT-PCR급면역조직화학등방법검측ERK1재대서간암세포주、대서원대간세포여12빈인간암급배대암방조직중적표체,응용선병독개도적세포외신호조절격매1소간우RNA(AdshERK1)감염대서간암세포주RH-35、H4IIE,관찰하조간암세포중ERK1표체후대기생물학특성적영향급하유암기인표체적변화.채용t검험진행통계학분석.결과 12빈인간암조직중,유11빈적ERK1표체교상응암방조직명현증다.ERK1재대서간암세포주RH-35、H4IIE중적표체교대서원대간세포표체상조(0.075±0.010、0.458±0.052비0.016±0.003,t=13.806、7.715,P균＜0.01).AdshERK1감염RH-35、H4IIE세포후,간암세포증식능력、극륭형성능력현저수억제.여AdshNC조상비,AdshERK1가명현감소RH-35세포중S기세포비례(18.55％±4.80％비4.39％±0.85％,t=-6.826,P＜0.01),인기G0/G1기세포주기조체(76.77％±0.93％비89.57％±1.26％,t=11.676,P＜0.01)；AdshERK1가현저억제H4IIE세포재라서피하성류능력[(0.603±0.284) mg비(0.051±0.074) mg,t=-5.340,P＜0.01].AdshERK1하조RH-35、H4IIE세포ERK1표체,병억제기하유암기인c-myc적표체.결론 AdshERK1유억제대서간암세포주RH-35화H4IIE증식、극륭형성급피하성류능력적작용,차해작용가능여억제ERK1하유종류상관기인c-myc적표체밀절상관.
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.