中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2013年
12期
925-929
,共5页
耿智敏%李波%王林%陈晨%李文智%郑见宝
耿智敏%李波%王林%陳晨%李文智%鄭見寶
경지민%리파%왕림%진신%리문지%정견보
索拉菲尼%肝细胞癌%肝星状细胞%肿瘤微环境
索拉菲尼%肝細胞癌%肝星狀細胞%腫瘤微環境
색랍비니%간세포암%간성상세포%종류미배경
Sorafenib%Hepatocellular carcinoma%Hepatic stellate cells%Tumor microenvironment
目的 研究索拉菲尼对肝星状细胞活性及活化的影响及其在肝脏肿瘤微环境中的作用机制.方法 通过LX2细胞和HepG2细胞共培养,观察LX2对HepG2细胞增殖的影响.通过MTT检测观察索拉菲尼对LX2增殖的影响.用不同浓度索拉菲尼干预LX2细胞,细胞组化检测LX2细胞α-SMA的表达;ELISA检测LX2上清液PDGF-BB和TGF-β1浓度变化;蛋白印迹检测LX2细胞ERK1、ERK2、Akt信号通路的表达.将不同浓度索拉菲尼干预的LX2细胞和HepG2细胞共培养24h,观察对HepG2细胞侵袭能力影响.结果 LX2和HepG2细胞共培养后实验组HepG2细胞明显较对照组多.经不同浓度索拉菲尼干预LX2细胞12h、24 h、36 h和48 h后,实验组细胞活性弱于对照组,具有浓度及时间依赖性.对照组α-SMA的表达强于实验组,具有浓度和时间依赖性.随着索拉菲尼药物浓度的升高及同一浓度下时间的延长,上清中PDGF-BB和TGF-β1的浓度降低.实验组与对照组中ERK1、ERK2、AKT的表达基本相同,但各自磷酸化状态的表达随着药物浓度的升高呈下降趋势.随着药物浓度的升高LX2诱导HepG2侵袭的能力逐渐减弱.结论 索拉菲尼可通过抑制α-SMA的表达、抑制肝星状细胞PDGF信号通路及下调上清液中的PDGF-BB和TGF-β1的表达,从而抑制HSC的活性及活化,继而抑制HepG2的增殖及侵袭.
目的 研究索拉菲尼對肝星狀細胞活性及活化的影響及其在肝髒腫瘤微環境中的作用機製.方法 通過LX2細胞和HepG2細胞共培養,觀察LX2對HepG2細胞增殖的影響.通過MTT檢測觀察索拉菲尼對LX2增殖的影響.用不同濃度索拉菲尼榦預LX2細胞,細胞組化檢測LX2細胞α-SMA的錶達;ELISA檢測LX2上清液PDGF-BB和TGF-β1濃度變化;蛋白印跡檢測LX2細胞ERK1、ERK2、Akt信號通路的錶達.將不同濃度索拉菲尼榦預的LX2細胞和HepG2細胞共培養24h,觀察對HepG2細胞侵襲能力影響.結果 LX2和HepG2細胞共培養後實驗組HepG2細胞明顯較對照組多.經不同濃度索拉菲尼榦預LX2細胞12h、24 h、36 h和48 h後,實驗組細胞活性弱于對照組,具有濃度及時間依賴性.對照組α-SMA的錶達彊于實驗組,具有濃度和時間依賴性.隨著索拉菲尼藥物濃度的升高及同一濃度下時間的延長,上清中PDGF-BB和TGF-β1的濃度降低.實驗組與對照組中ERK1、ERK2、AKT的錶達基本相同,但各自燐痠化狀態的錶達隨著藥物濃度的升高呈下降趨勢.隨著藥物濃度的升高LX2誘導HepG2侵襲的能力逐漸減弱.結論 索拉菲尼可通過抑製α-SMA的錶達、抑製肝星狀細胞PDGF信號通路及下調上清液中的PDGF-BB和TGF-β1的錶達,從而抑製HSC的活性及活化,繼而抑製HepG2的增殖及侵襲.
목적 연구색랍비니대간성상세포활성급활화적영향급기재간장종류미배경중적작용궤제.방법 통과LX2세포화HepG2세포공배양,관찰LX2대HepG2세포증식적영향.통과MTT검측관찰색랍비니대LX2증식적영향.용불동농도색랍비니간예LX2세포,세포조화검측LX2세포α-SMA적표체;ELISA검측LX2상청액PDGF-BB화TGF-β1농도변화;단백인적검측LX2세포ERK1、ERK2、Akt신호통로적표체.장불동농도색랍비니간예적LX2세포화HepG2세포공배양24h,관찰대HepG2세포침습능력영향.결과 LX2화HepG2세포공배양후실험조HepG2세포명현교대조조다.경불동농도색랍비니간예LX2세포12h、24 h、36 h화48 h후,실험조세포활성약우대조조,구유농도급시간의뢰성.대조조α-SMA적표체강우실험조,구유농도화시간의뢰성.수착색랍비니약물농도적승고급동일농도하시간적연장,상청중PDGF-BB화TGF-β1적농도강저.실험조여대조조중ERK1、ERK2、AKT적표체기본상동,단각자린산화상태적표체수착약물농도적승고정하강추세.수착약물농도적승고LX2유도HepG2침습적능력축점감약.결론 색랍비니가통과억제α-SMA적표체、억제간성상세포PDGF신호통로급하조상청액중적PDGF-BB화TGF-β1적표체,종이억제HSC적활성급활화,계이억제HepG2적증식급침습.
Objective To investigate the effects and mechanisms of sorafenib on hepatic stellate cell viability and activation in the microenvironment of liver tumor.Methods The effects of LX2 cells on HepG2 cell proliferation were observed by coculture of LX2 and HepG2 cells.MTT assay was used to observe the effects of sorafenib on LX2 proliferation,and expression of α-smooth muscle actin (α-SMA) was measured immunocytochemically in LX2 cells treated with different concentrations of sorafenib.Changes in PDGF-BB and TGF-β1 concentrations were detected in LX2 supernatant using ELISA.Expression of ERK1,ERK2,and AKT signaling pathways were measured using Western blot.Furthermore,LX2 cells were cocultured with HepG2 cells for 24 hours to observe their effects on the invasive ability of HepG2 cells.Result After coculture of LX2 and HepG2 cells,HepG2 cells increased in the experimental group more than those of in the control group.After treatment with various concentrations of sorafenib for 12,24,36 or 48 hours,the viability of treated LX2 cells was lower than the controls in a concentration-and time-dependent manner.As sorafenib concentration and time of exposure increased,α-SMA expression became weaker in treated cells.PDGF-BB and TGF-β1 concentrations decreased with higher sorafenib concentrations and with longer exposure under the same concentration.ERK1,ERK2 and Akt expression was identical between treated and control groups,but their phosphorylated expression decreased with increased concentrations of sorafenib.The invasive ability of HepG2 cells induced by LX2 gradually decreased as sorafenib concentrations increased.Conclusions Sorafenib suppressed α-SMA expression,inhibited PDGF-dependent signaling pathways in HSCs,downregulated PDGF-BB and TGF β1 expression in the supernatant of HSCs,and restrained the viability and activation of HSCs.Sorafenib treatment therefore resulted in suppressed proliferation and invasion of HepG2 cells.
