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.     

蛋白激酶C-α反义核酸对人肝癌细胞HepG2体外增殖及凋亡的影响

目的　探讨蛋白激酶 (PK )C α反义寡核苷酸 (asODN)对人肝癌细胞HepG2体外增殖及凋亡的影响。方法　用RPMI 164 0体外培养和传代HepG2细胞 ,以脂质体 (LP)介导不同浓度PKC αasODN转染人肝癌细胞HepG2 ,四甲基偶氮唑蓝 (MTT)比色法检测其生长指数 (GI) ,软琼脂克隆形成法检测其克隆形成率 ,流式细胞术 (FCM )检测其凋亡率。结果　 0 .10～ 1.0 0 μmol/L的PKC αasODN能显著降低HepG2细胞GI(P <0 .0 5 ) ;0 .0 5～ 1.0 0 μmol/L的PKC αasODN能显著降低其软琼脂克隆形成率 (P <0 .0 1) ,两者均具有量效依赖关系。 0 .5 0～ 1.0 0 μmol/L的PKC αa sODN可使HepG2细胞发生显著凋亡 (P <0 .0 1)。结论　通过PKC αasODN阻断人肝癌细胞HepG2信号转导分子PKC α ,可有效抑制HepG2体外生长增殖 ,并可诱导其凋亡     

IGF-Ⅱ对阿霉素诱导人肝癌细胞HepG2凋亡的拮抗作用研究

目的 研究胰岛素样生长因子Ⅱ(insulin-like growth factor Ⅱ,IGF-Ⅱ)对阿霉素(adriamycin,ADM)诱导的人肝癌细胞HepG2凋亡的影响及其与凋亡抑制蛋白survivin表达的关系.方法 实验分组:A组:空白对照组;B组:200 ng/ml ADM组;C组:2 n/ml IGF-Ⅱ+200 ng/ml ADM组;D组:20 ng/ml IGF-Ⅱ+200 ng/ml ADM组;E组:200 ng/ml IGF-Ⅱ+200 ng/ml ADM组,分别处理HepG2细胞48 h后,应用MTF检测各组细胞活力,流式细胞仪检测细胞的凋亡率,Western blot检测细胞中survivin蛋白的表达.结果 A、B、C、D、E组的HepG2细胞活力分别为:0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013、0.304±0.019,凋亡率分别为:6.9%±1.3%、35.4%±2.1%、31.2%±2.2%、26.4%±1.7%、21.7%±1.9%,survivin与β-actin的比值分别为:0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033、0.469±0.031,结果显示IGF-Ⅱ+ADM组的HepG2细胞活力较ADM组明显好转(P＜0.01),凋亡率显著降低(P＜0.01),IGF-Ⅱ+ADM组的HepG2细胞中survivin蛋白的表达较ADM组明显增高(P＜0.01),且IGF-Ⅱ的上述作用随IGF-Ⅱ浓度的增高而增强(P＜0.05).结论 IGF-Ⅱ能够上调HepG2细胞中survivin蛋白的表达,拮抗ADM诱导的HepG2细胞凋亡.     

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.     

COX-2反义RNA抑制肝癌细胞增殖的实验研究

目的 研究COX-2反义RNA对肝癌细胞增殖抑制的作用及其机制,探讨肝癌治疗的新途径.方法 人工合成的COX-2反义RNA片段及无关对照空质粒经脂质体包裹后作用CBRH7919细胞.通过MTT法、细胞周期分析、RT-PCR及裸鼠体内接种等方法测定细胞体内外增殖的变化.结果 经COX-2反义RNA处理的CBRH7919细胞与对照组CBRH7919细胞相比,体外增殖速度减慢(细胞增殖抑制率达78%)、DNA合成受抑(S期细胞数为34.8%vs59.9%),细胞周期G_0/G.比例明显提高,裸鼠体内成瘤率下降(25%vs100%).而凋亡相关基因表达并无明显差异(P>0.05).结论 COX-2反义RNA可有效抑制肝癌细胞的体内外生长增殖,可用于实验性肿瘤基因治疗研究.