普伐他汀对肝癌HepG2细胞增殖的抑制作用

目的探讨HMG-CoA还原酶抑制剂(Statins)在体内外抑制肝细胞癌增殖和侵犯的作用及其机制。方法以不同浓度的普伐他汀(Pr)作用于培养的HepG2细胞,用噻唑蓝比色试验检测细胞的增殖活性,并观察细胞的形态学变化。建立HepG2细胞裸小鼠皮下移植瘤模型,腹腔注射浓度为5 g／L的Pr,每只每日10 ml／kg体重。观察肿瘤的生长情况,实验结束时切取肿瘤称重,检测血中AFP水平。肿瘤标本石蜡切片,免疫组织化学检测CyclinD1和FⅧRA的表达。结果 Pr作用96 h后明显抑制了HepG2细胞的生长,细胞出现明显的形态学改变。Pr也抑制裸小鼠皮下移植型肝癌的生长,Pr组和对照组瘤重分别是(0．246±0．129)g和(0．376±0．102)g(P< 0．05),抑瘤率34．6％。两组的AFP表达差异有统计学意义[(2．657±1．465)μg／L比(4．206± 1．204)μg／L,P<0．05]。Cydin D1在肿瘤间质的成纤维细胞表达,Pr组的积分吸光度(OPTID)为 219．40±24．00,对照组为451．54±47．97(P<0．05)。两组的微血管密度(MVD)分别为27．43± 6．83和41．22±6．26(P<0．05)。结论 Pr能抑制肝癌细胞的增殖,其机制主要是抑制肿瘤的血管形成和肿瘤基质中的成纤维细胞的增生。     

5-氟尿嘧啶缓释微粒对肝癌HepG2细胞的抑制作用

目的 研究5-氟尿嘧啶(5-fluorouracil,5-FU)缓释微粒对肝癌HepG2细胞的抑制作用.方法 MTT法检测5-FU缓释微粒、5-FU裸药和缓释微粒载体(聚L-乳酸)对人肝癌HepG2细胞的抑制作用,采用Hochest染色和流式细胞仪检测细胞凋亡.结果 5-Fu缓释微粒组细胞存活率随着时间的延长而降低,凋亡细胞数目也随之增加.5-FU裸药组第1天的抑制作用最强,有大量细胞凋亡,随着作用时间的延长,细胞存活率缓慢上升,第21、28天的细胞存活率高于5.FU缓释微粒组.缓释微粒载体组细胞凋亡均较少.3组比较差异有统计学意义(F=3163.52,128.47,P＜0.01).结论 植入用5-FU缓释微粒可以在28 d内持续释放,并且以时间依赖性方式抑制肝癌HepG2细胞生长和诱导其凋亡增加,明显优于5-FU裸药.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

目的 研究反义HCCR-2真核表达载体对肝癌HepG2细胞增殖及凋亡的影响.方法 构建反义HCCR-2真核表达载体(反义载体组),转染肝癌HepG2细胞,G418筛选阳性克隆,同样方法获得空载体pIRES2-EGFP稳定表达的细胞株(空载体组),取肝癌HepG2细胞为对照(肝癌HepG2组),用MTT法、流式细胞仪、透射电镜观察反义HCCR-2转染前后肝癌HepG2细胞生长曲线、细胞周期、细胞凋亡及细胞形态的变化.采用单因素方差分析和χ2检验比较各组差异.结果 反义载体组、空载体组、肝癌HepG2组HCCR-2 mRNA表达水平分别为0.39±0.04、0.62±0.06、0.72±0.03,3组比较差异有统计学意义(F=43.701,P<0.05);细胞凋亡率分别为13.30%、2.51%、2.07%,反义载体组与空载体组、肝癌HepG2组比较,差异有统计学意义(χ2=6.793,8.721,P<0.05);反义载体组细胞生长减慢,阻滞于G0/G1期.结论 HCCR-2反义核酸真核表达载体能抑制HCCR-2 mRNA的表达,促进细胞凋亡,HCCR-2蛋白可能参与肝癌细胞的周期调控,并与细胞的生长增殖有关.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

HCCR-2反义核酸对肝癌HepG2细胞的作用

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.     

化学治疗药物对肝癌细胞增殖抑制作用的观察

目的 评价化学治疗药物（下称化疗药）在肝癌治疗中的作用和地位。方法 采用MTT法测定5-Fu、顺铂和4-PA对肝癌细胞增殖的抑制作用。结果 （1）5-Fu、顺铂两药分别单剂使用,在给药后0.5h肝癌细胞增殖h后肝癌细胞增殖均受到抑制;对氯苯乙酸在给药后0.5h肝癌细胞增殖即被抑制。（2）肝癌细胞抑制率随化疗药的作用时间延长而增加,48-72h达50%以上的抑制率。（3）化疗药对肝癌细胞的作用早期,癌细胞的生长有返跳活跃阶段。但总体上癌细胞伯生长被抑制。结论 连续用药可很好地抑制肝癌细胞增殖,提高恶性肿瘤化学药物治疗的效果。     

siRNA对肝癌细胞中特异性异常表达FGFR3基因的抑制效应研究

目的设计和筛选有效抑制纤维母细胞生长因子受体3基因（FGFR3）表达的siRNA序列,构建相应表达载体,研究其对肝癌细胞HepG2生物学特性的影响。方法根据FGFR3序列,设计3条siRNA序列及1条对照序列：以pRNAT-U 6．1／Neo为载体,并以载体中同时构建的绿色荧光蛋白（GFP）基因编码序列为转染效率内参照;采用脂质体法将pRNAT-U 6．1／Neo-siRNA转染肝癌细胞HepG2;以实时荧光定量PCR（Real time PCR）检测FGFR3的mRNA表达变化,Western印迹法检测FGFR3蛋白质表达变化;[^3H]胸腺嘧啶脱氧核苷掺入法（^3H-Tdr）和细胞克隆形成法分析DNA合成变化及细胞生长能力。结果所设计合成的3条siRNA序列中均能抑制FGFR3的表达,对mRNA表达抑制率约为74．04％～87．25％,在蛋白质水平表达抑制率为19．65％～90．40％。转染siRNA后,HepG2的细胞克隆形成能力和DNA合成能力明显受到抑制。结论siRNA能够有效抑制肝癌细胞FGFR3的表达,并继而抑制肝癌细胞的DNA合成和生长,能够为肝癌基因表达调控研究提供新的靶基因。     

甲基强的松龙对体外肝癌HepG2细胞生长的影响

目的 探讨甲基强的松龙（MP）对肝癌HepG2细胞生长的影响及相关机制。方法 MTT法测定不同浓度MP对HepG2细胞增殖的影响;细胞分为6个处理组：对照组、MPⅠ组（10 ug/l）、MPⅡ组（20 ug/l）、MPIII组（50 ug/l）、MPIV组（500 ug/l）、MPV组（5000 ug/l）;流式细胞仪测定MP对HepG2细胞周期和凋亡情况的作用;荧光定量PCR法测定MP对HepG2细胞VEGF mRNA表达的影响;ELISA法测定细胞培养上清液VEGF浓度。结果 50-5000ug/l的MP作用72h,能促进HepG2细胞的增殖;MP对HepG2细胞周期和凋亡无影响;不同浓度的MP作用72h后,HepG2细胞VEGF mRNA的转录和蛋白表达有显著性差异（P＜0.01）,MP在500ug/l和5000ug/l浓度时, VEGF mRNA表达量分别为（7.293±0.409）拷贝数/ul对数值和（8.231±0.216）拷贝数/ul对数值,高于对照组（4.761±0.624）拷贝数/ul对数值,结果有显著性差异（P＜0.01）,VEGF浓度分别为（11.153±1.552）ug/l和（11.456±1.517）ug/l,高于对照组（7.362±0.510）ug/l,结果有显著性差异（P＜0.01）;。结论 高浓度的MP在体外能促进肝癌HepG2细胞增殖,上调其VEGF mRNA的转录和翻译。     

缺氧对肝癌细胞系HepG2表达VEGF的影响

目的研究缺氧条件下肝癌细胞系HepG2表达血管内皮生长因子（VEGF）的变化。方法分别在缺氧（缺氧组）和有氧（对照组）条件下培养HepG2细胞．采用免疫组织化学染色、MTT比色、细胞计数和ELISA方法．检测2组VEGF表达、HepG2细胞生长情况和培养上清液中VEGF含量。结果缺氧培养可抑制HepG2细胞的生长．缺氧培养48和72h后．HepG2细胞的生长速度明显低于对照组相应时相（P〈0．05．P〈0．01）；缺氧培养24和48h后．HepG2细胞上清液中VEGF含量明显高于对照组相应时相（P〈0．05。P〈0．01）。结论缺氧可以促进肝癌细胞HepG2表达分泌VEGF．这可能是经导管肝动脉化疗栓塞术后VEGF高表达的原因。     

缺氧对肝癌细胞内β-链蛋白表达的影响

目的 探讨缺氧对肝癌细胞内β-链蛋白(β-catenin)表达的影响及机制.方法 分别利用100 μmol/L氯化钴和肝癌原位移植+肝动脉结扎建立肝癌细胞体内、外缺氧模型.实时定量逆转录-聚合酶链反应(RT-PCR)、Western blot、免疫荧光和免疫组织化学法分析缺氧对PLC/PRF/5、Hep3B、HepG2和MHCC97 4种肝癌细胞内β-catenin mRNA和蛋白表达的影响.结果 缺氧不影响肝癌细胞β-catenin mRNA表达,但增加HepG2和PLC/PRF/5细胞内β-catenin总蛋白水平(＞2.3倍),促进MHCC97及Hep3B细胞β-catenin的细胞内易位(胞质和/或胞核,＞1.7倍).这与缺氧诱导的肝癌细胞内糖原合成酶3β下调,β-catenin蛋白降解抑制有关.结论 缺氧促进肝癌细胞内β-catenin蛋白表达和细胞内转移,增加细胞恶性潜能.     

siRNA靶向沉默TET2基因对人肝癌细胞增殖和凋亡的影响

目的:通过siRNA靶向沉默TET2(ten-eleven translocation 2)基因,以研究TET2基因对于人肝癌细胞增殖和凋亡的影响并初步探讨其可能机制。方法:RT-PCR和Western印迹法检测人肝癌细胞株和正常人肝星状细胞LO2中TET2表达情况。siRNA转染入Bel-7404和SMMC-7721肝癌细胞内,并设置阴性对照组(NC组)。靶向沉默TET2基因后,CCK-8法检测肝癌细胞增殖能力,FITC AnnexinⅤ-Apoptosis Detection KitⅠ检测肝癌细胞的凋亡;Western印迹法检测下游信号通路中Caspase-3和Caspase-8蛋白表达。结果:与正常人肝星状细胞LO2相比,TET2基因在人肝癌细胞中高表达。siRNA转染入SMMC-7721和Bel-7404细胞后,TET2基因的表达显著降低。与NC组相比,转染48 h后肝癌细胞的增殖水平显著降低(P<0.05)。肝癌细胞的凋亡水平率显著升高,下游信号通路中,Capase-3蛋白和Capase-8蛋白的表达量显著上调。结论:siRNA靶向沉默TET2基因可通过促进肝癌细胞凋亡的发生抑制其增殖。     

羟基脲对肝癌细胞HepG2中GADD45β表达影响及其可能机制

目的 初步明确肝癌细胞中GADD45β基因近端启动子序列,探索羟基脲对人肝癌细胞HepG2的GADD45β表达影响及可能机制.方法 体外合成GADD45β近端启动子序列群(-618～-314),构建荧光素表达质粒,转染肝癌细胞株HepG2,根据启动子活性强度结合数据库分析存在的转录调节因子结合位点;以实时荧光定量PCR比较羟基脲作用前后HepG2细胞GADD45β表达,并进一步比较羟基脲对GADD45β启动子活性的调控作用、分析羟基脲对HepG2的抑制效应,并通过Caspase-8、Caspase-9和Caspase-3的表达变化测定凋亡的发生和发展.结果 GADD4518近端启动子中含有3个NF-кB(-602/-593、-581/-572、-537/-528)和1个E2F-1(-470/-436)转录调节因子与启动子结合位点;羟基脲能明显诱导HepG2中GADD45β的表达,并呈现出剂量-效应的正相关关系,同时NF-кB和E2F-1启动子均明显增强.羟基脲能够明显抑制HepG2的DNA合成能力和细胞克隆形成能力,同时羟基脲能迅速启动HepG2凋亡的发生和发展.结论 羟基脲能明显诱导肝癌细胞中特异性缺失的GADD45β基因表达,增强转录调节因子的表达水平是其可能的作用机制.     

肝癌患者转化生长因子—β1及其受体的表达和意义

目的 探讨肝癌患者转化生长因子－β１（ＴＧＦ－β１）及其受体的表达情况和意义。方法 酶联免疫吸附法检测２７例行胆囊切除术者（对照组）和３６例肝癌患者（肝切除组）肝切除手术前后血清ＴＧＦ－β１浓度;免疫组织化学法检测３６例肝癌和癌周组织标本中相关受体的表达情况。结果 肝癌患者血清ＴＧＦ－β１浓度较对照组显著升高（Ｐ〈０．０５）;患者肝切除术后血清ＴＧＦ－β１浓度较术前有显著升高（Ｐ〈０．０１）,而胆     

损伤相关模式分子对人肝癌HepG2细胞增殖能力的影响

目的 观察在DAMPs诱导下人肝癌HepG2细胞增殖能力的变化.方法 将HepG2细胞分为对照组和实验组（10、20、40、80 μl DAMPs处理的HepG2细胞）;MTT比色法检测HepG2细胞的增殖能力;实时荧光定量PCR法检测IL-6 mRNA表达的变化;Western Blot法检测HepG2细胞IL-6蛋白的表达情况.结果 HepG2细胞随着DAMPs剂量的递增及时间的延长增殖能力逐渐增强,呈现明显的量效-时效关系,在剂量40 μl,作用时间36 h时细胞增殖能力达到最强,差异有统计学意义（P〈0.01）;选取作用时间为36 h,随着DAMPs剂量的递增,实时定量PCR法检测到HepG2细胞IL-6 mRNA分别为95.55±4.47,171.80±6.60,453.30±14.47,610.59±12.70,441.04±18.91,差异有统计学意义（P〈0.01）;Western Blot法检测HepG2细胞IL-6蛋白的表达分别为1.47、2.07、2.74、3.44、3.00,差异有统计学意义（P〈0.01）.结论 DAMPs在一定剂量及时间内促进人肝癌HepG2细胞增殖,并且呈现明显的量效-时效关系.