OBJECTIVE Hepatocel ular carcinoma(HCC)is the most common cause of cancer-related mortality,with high incidence rates,robust metastatic propensity and acquired resistance to therapy.Metformin,an extensively prescribed...OBJECTIVE Hepatocel ular carcinoma(HCC)is the most common cause of cancer-related mortality,with high incidence rates,robust metastatic propensity and acquired resistance to therapy.Metformin,an extensively prescribed and well-tolerated first-linetherapeutic drug for type 2 diabetes mellitus,has recently been identified as a potential and attractive anticancer adjuvant drug combined with chemotherapeutics to improve treatment efficacy and lower doses.Curcumin,a botanical extracts,has been shown antitumorigenic properties.This study aims to investigate the combinational effect of metformin and curcumin on inbibition of tumor growth and metastasis in Hep G2 cells and the possible underlying mechanisms.METHODS The cell proliferation was determined by MTT,CCK-8 and colony formation assay.The protein expression was detected by Western blotting.Activity of MMP-2 and MMP-9 was estimated by gelatin zymography.Flow cytometry analysis was used to evaluate the influence of metformin and curcumin on cell cycle arrest and apoptosis,and morphology observation of apoptosis was detected by Hoechst33342.Scratch and transwell assay was performed to detect the cell migration and invasion.The suppression of this combination therapy oncapillary tube formation was detected by tube formation assay.RESULTS Combination of metformin and curcumin induced stronger inhibition on Hep G2 cells proliferation than monotherapywhich related to induction of cell cycle arrest in G2/M phase and apoptosis through regulation of the protein expression of cyclin B and Bcl-2/Bax.Moreover,the co-treatment of metformin with curcumin exerted an enhanced inhibitory effect on Hep G2 cell metastasis and synergistically inhibited the tube formation of HUVEC cells.The suppression of PI3K/AKT/m TOR pathway and inhibition the protein expression of STAT3,MMP9,MMP2 and VEGF might involve in this synergistic effects of combination treatment.CONCLUSION Combination of metformin and curcumin inhibited Hep G2 cells proliferationmore effectively than monotherapy and synergistically induced a greater inhibition on migration and invasion of Hep G2 cells.展开更多
目的:鉴定人肝母细胞瘤HepG2细胞中响应DNA损伤的转运RNA衍生的小RNA(transfer RNAderived small RNA,tsRNA)的表达特征,并研究其潜在功能。方法:本研究基于配对的HepG2细胞和敲除TP53基因的HepG2细胞,采用阿霉素(adriamycin,ADR)成功构...目的:鉴定人肝母细胞瘤HepG2细胞中响应DNA损伤的转运RNA衍生的小RNA(transfer RNAderived small RNA,tsRNA)的表达特征,并研究其潜在功能。方法:本研究基于配对的HepG2细胞和敲除TP53基因的HepG2细胞,采用阿霉素(adriamycin,ADR)成功构建DNA损伤的细胞模型,并进行小非编码RNA的转录组分析,系统地鉴定一批响应ADR并参与p53调节的tsRNA,利用京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)进行了功能富集分析。此外,经沉默目标tsRNA基因表达后,通过CCK-8实验和平板集落形成实验初步证实了目标tsRNA在HepG2细胞模型中的生物学功能。结果:DNA损伤可诱导一批参与p53调节的tsRNA,其中tRF-5-1(tRF-5_tRNA-Gly-TCC-2-1)和tRF-i-1(tRF i_tRNA-Tyr-GTA-11-1)在HepG2细胞中的表达上调最为显著(P<0.05)。沉默tRF-5-1或tRF-i-1基因可抑制HepG2细胞的增殖活力(P<0.05)。结论:HepG2细胞模型中可以鉴定一组响应DNA损伤的tsRNA,且tsRNA可以促进HepG2细胞的增殖活力,提示tsRNA在肝脏细胞的恶性增殖中可能扮演重要角色。展开更多
文摘OBJECTIVE Hepatocel ular carcinoma(HCC)is the most common cause of cancer-related mortality,with high incidence rates,robust metastatic propensity and acquired resistance to therapy.Metformin,an extensively prescribed and well-tolerated first-linetherapeutic drug for type 2 diabetes mellitus,has recently been identified as a potential and attractive anticancer adjuvant drug combined with chemotherapeutics to improve treatment efficacy and lower doses.Curcumin,a botanical extracts,has been shown antitumorigenic properties.This study aims to investigate the combinational effect of metformin and curcumin on inbibition of tumor growth and metastasis in Hep G2 cells and the possible underlying mechanisms.METHODS The cell proliferation was determined by MTT,CCK-8 and colony formation assay.The protein expression was detected by Western blotting.Activity of MMP-2 and MMP-9 was estimated by gelatin zymography.Flow cytometry analysis was used to evaluate the influence of metformin and curcumin on cell cycle arrest and apoptosis,and morphology observation of apoptosis was detected by Hoechst33342.Scratch and transwell assay was performed to detect the cell migration and invasion.The suppression of this combination therapy oncapillary tube formation was detected by tube formation assay.RESULTS Combination of metformin and curcumin induced stronger inhibition on Hep G2 cells proliferation than monotherapywhich related to induction of cell cycle arrest in G2/M phase and apoptosis through regulation of the protein expression of cyclin B and Bcl-2/Bax.Moreover,the co-treatment of metformin with curcumin exerted an enhanced inhibitory effect on Hep G2 cell metastasis and synergistically inhibited the tube formation of HUVEC cells.The suppression of PI3K/AKT/m TOR pathway and inhibition the protein expression of STAT3,MMP9,MMP2 and VEGF might involve in this synergistic effects of combination treatment.CONCLUSION Combination of metformin and curcumin inhibited Hep G2 cells proliferationmore effectively than monotherapy and synergistically induced a greater inhibition on migration and invasion of Hep G2 cells.
文摘目的:观察化瘀祛痰方(Huayu-Qutan formula,HYQT)含药血清对油酸诱导的HepG2细胞脂质损伤及内质网应激(endoplasmic reticulum stress,ERS)的影响,探讨HYQT防治脂代谢紊乱的可能机制。方法:以不同终浓度的油酸诱导HepG2细胞,CCK8法、油红O染色和ELISA法筛选油酸诱导及HYQT含药血清干预的最佳浓度及时间。将HepG2细胞分为对照(control)组、模型(model)组、HYQT组、毒胡萝卜素(thapsigargin,Tg;ERS激动剂)组和Tg+HYQT组。油红O染色观察细胞脂质沉积情况;ELISA法检测细胞甘油三酯(triglyceride,TG)和游离脂肪酸(free fatty acid,FFA)含量变化;透射电镜观察细胞内质网结构变化;RT-qPCR和Wes全自动蛋白定量分析系统检测各组细胞葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)、固醇调节元件结合蛋白1c(sterol regula⁃tory element binding protein 1c,SREBP1c)、乙酰辅酶A羧化酶1(acetyl coenzyme A carboxylase 1,ACC1)、脂肪酸合成酶(fatty acid synthase,FAS)和硬脂酰辅酶A去饱和酶l(stearoyl coenzyme A desaturase 1,SCD1)的mRNA和蛋白表达情况。结果:1000μmol/L油酸诱导HepG2细胞脂质沉积效果明显,10%含药血清作用48 h作为后续实验的相对最佳浓度和时间。与control组比较,model组胞浆中橘红色脂滴形成明显增多,内质网扩张,发生ERS,细胞核固缩;TG和FFA含量明显增多(P<0.01);GRP78、SREBP1c、ACC1、FAS和SCD1 mRNA及蛋白表达量显著增高(P<0.01);与model组比较,HYQT含药血清干预后细胞内橘红色脂滴明显减少,内质网形态基本恢复正常,TG和FFA含量明显减少(P<0.01),ERS及脂肪酸从头合成相关因子的mRNA和蛋白表达量显著降低(P<0.01);加入Tg后细胞内橘红色脂滴形成更多,内质网扩张程度更明显,内质网形态迥异,大小不一,呈空泡状改变,部分可见融合成簇现象,TG和FFA含量增多(P<0.01),ERS及脂肪酸从头合成相关因子的mRNA和蛋白表达量增高(P<0.01);与Tg组比较,Tg+HYQT组细胞内橘红色脂滴减少,内质网扩张程度减轻,空泡体积减小和数量减少;TG和FFA含量明显减少(P<0.01),ERS及脂肪酸从头合成相关因子的mRNA和蛋白表达量显著降低(P<0.01)。结论:HYQT含药血清可以减轻油酸诱导的HepG2细胞脂质损伤,其机制可能与抑制ERS介导的脂肪酸从头合成有关。
文摘目的:鉴定人肝母细胞瘤HepG2细胞中响应DNA损伤的转运RNA衍生的小RNA(transfer RNAderived small RNA,tsRNA)的表达特征,并研究其潜在功能。方法:本研究基于配对的HepG2细胞和敲除TP53基因的HepG2细胞,采用阿霉素(adriamycin,ADR)成功构建DNA损伤的细胞模型,并进行小非编码RNA的转录组分析,系统地鉴定一批响应ADR并参与p53调节的tsRNA,利用京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)进行了功能富集分析。此外,经沉默目标tsRNA基因表达后,通过CCK-8实验和平板集落形成实验初步证实了目标tsRNA在HepG2细胞模型中的生物学功能。结果:DNA损伤可诱导一批参与p53调节的tsRNA,其中tRF-5-1(tRF-5_tRNA-Gly-TCC-2-1)和tRF-i-1(tRF i_tRNA-Tyr-GTA-11-1)在HepG2细胞中的表达上调最为显著(P<0.05)。沉默tRF-5-1或tRF-i-1基因可抑制HepG2细胞的增殖活力(P<0.05)。结论:HepG2细胞模型中可以鉴定一组响应DNA损伤的tsRNA,且tsRNA可以促进HepG2细胞的增殖活力,提示tsRNA在肝脏细胞的恶性增殖中可能扮演重要角色。
