摘要
目的明确营养剥夺促进肝癌细胞侵袭的机制。方法以Hank’s平衡盐缓冲液对肝癌细胞系HepG2和BEL7402细胞进行营养剥夺培养,同时构建转染针对自噬基因Atg3的siRNA(siRNA-Atg3),观察营养剥夺在Atg3基因沉默前后对肝癌细胞自噬活性和侵袭能力的影响,分析自噬活性与侵袭能力之间的关系。结果平衡盐缓冲液较对照完全培养液能明显诱导肝癌细胞HepG2和BEL7402内自噬体形成[(93±5)vs.(40±6)/高倍视野,P<0.05;(83±4)vs.(36±4)/高倍视野,P<0.05]及LC3蛋白的表型转换,抑制P62蛋白表达,同时显著增加HepG2和BEL7402细胞的侵袭数[(60 875±1 051)vs.(15 812±515),P<0.05;(59 750±1 571)vs.(15 500±645),P<0.05]。转染siRNA-Atg3可沉默细胞Atg3表达并显著对抗营养剥夺诱导的LC3蛋白表型转换和P62蛋白表达下调,HepG2和BEL7402肝癌细胞的侵袭数较未转染细胞也明显减少[(17 438±427)vs.(59 875±520),P<0.05;(17 250±408)vs.(59 500±540),P<0.05]。结论自噬可能是营养剥夺条件下促进肝癌细胞侵袭性增加的主要机制。
Objective To elucidate the mechanism by which nutritional deprivation promotes invasion of hepatocellular carcinoma（HCC）cells.Methods HepG2 and BEL7402 cells were starved in Hank＇s balanced salt solution（HBSS）.siRNA that targeted autophagy gene,Atg3,was constructed and transfected to observe its effect on autophagic activity and invasion of HCC cells under starvation.The relationship was further analyzed between autophagy and invasion of HCC cells under starvation.Results As compared with control complete medium,HBSS notably increased autophagosome formation [（93±5）vs.（40±6）/high field,P0.05;（83±4）vs.（36±4）]/high field,P0.05],induced phenotype transition of LC3 and decreased expression of P62 in HepG2 and BEL7402 cells.At the same time,HBSS significantly promoted invasion of HepG2 and BEL7402 cells [（60 875±1 051）vs.（15 812±515）,P0.05;（59 750±1 571）vs.（15 500±645）,P0.05].However,silence of Atg3 by siRNA significantly abrogated above phenotype transition of LC3 and decrease in P62 expression,meanwhile the invasion number of HepG2 and BEL7402 cells was also reduced under nutritional deprivation in contrast to non-transfected cells [（17 438±427）vs.（59 875±520）,P0.05;（17 250±408）vs.（59 500±540）,P0.05].Conclusion Nutritional deprivation-promoted invasion of HCC cells may predominantly depend on autophagy.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第5期513-517,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省自然科学基金资助项目(No.2011CDB103)
关键词
肝癌细胞
自噬
营养剥夺
侵袭
hepatocellular carcinoma cells
autophagy
nutritional deprivation
invasion
