AIM: To investigate the potential roles of enhancer of zeste homolog2(EZH2), Bmi-1 and mi R-203 in cell proliferation and invasion in hepatocellular carcinoma(HCC) cell line Hep3 B.METHODS: A total of 73 patients who ...AIM: To investigate the potential roles of enhancer of zeste homolog2(EZH2), Bmi-1 and mi R-203 in cell proliferation and invasion in hepatocellular carcinoma(HCC) cell line Hep3 B.METHODS: A total of 73 patients who underwent surgical resection at Fuzong Clinical Medical College of Fujian Medical University were enrolled in this study. Hep3 B cells were cultivated in RPMI 1640 medium supplemented with 10% fetal bovine serum at 37?℃. Vectors that containing c DNA of the EZH2 gene or mi R-203 targeted sh RNA plasmid were constructed, and then transfected into Hep3 B cells. The m RNA expression of mi R-203, EZH2, and Bmi-1 was analyzed using quantitative real-time polymerase chain reaction analysis, and the protein levels of EZH2 and Bmi-1 were detected by Western blot analysis. Effect of EZH2 or mi R-203 on cell proliferation was observed by methyl thiazolyl tetrazolium assay, and cell apoptosis was assessed using flow cytometry. Besides, effect of EZH2 or mi R-203 on tumor cell invasion was detected using Transwell assay.RESULTS: The m RNA levels of EZH2 and Bmi-1 in HCC tissues and in Hep3 B cells were significantly higher compared with those in normal samples(P < 0.01), while mi R-203 level was significantly lower in HCC tissues(P < 0.01). Hep3 B cells transfected with EZH2-sh RNA or mi R-203-sh RNA showed lower expression levels of EZH2 and Bmi-1(P < 0.05). Compared with controls, Hep3 B cells transfected with EZH2-sh RNA had relative slow cell proliferation, indicating that low expression of EZH2 and Bmi-1 and overexpression of mi R-203 could inhibit Hep3 B cell proliferation(P < 0.05). The average apoptosis rate of Hep3 B cells transfected with EZH2-sh RNA vector was about 18.631%, while that of Hep3 B cells transfected with sh RNA vector was about 5.33%, suggesting that EZH2 was down-regulated by transfecting with EZH2-sh RNA, and the down-regulated EZH2 contributed to the cell apoptosis. Low expression of EZH2 and Bmi-1 and overexpression of mi R-203 could reduce Hep3 B cell invasion(P < 0.05).CONCLUSION: Our study suggests that EZH2 and Bmi-1 are up-regulated while mi R-203 is downregulated in Hep3 B cells. Mi R-203 may contribute to the metastasis and enhance apoptosis of HCC cells by regulating EZH2 and Bmi-1. Our study may provide a theoretical basis for metastasis of HCC and targeted therapy of HCC.展开更多
In order to study the effect of 5, 6-Dichloro-l-13-D-ribofuranosyl-benzimidazole （DRB） on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were trea...In order to study the effect of 5, 6-Dichloro-l-13-D-ribofuranosyl-benzimidazole （DRB） on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry （FCM） and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was （0.68±0.19）%, （1.95±0.12）%, （8.51±0.26）%, （11.26±0.17）% and （14.99±0.32）%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.展开更多
目的:研究小干扰RNA(small interfering RNA,si RNA)抑制亲环素A(cyclopilin A,CypA)的表达对喉癌细胞株Hep-2增殖和侵袭迁移的影响及相关机制。方法:实验分为3组:化学合成的靶向CypA的si RNA转染喉癌细胞Hep-2(CypA si RNA组),转染阴...目的:研究小干扰RNA(small interfering RNA,si RNA)抑制亲环素A(cyclopilin A,CypA)的表达对喉癌细胞株Hep-2增殖和侵袭迁移的影响及相关机制。方法:实验分为3组:化学合成的靶向CypA的si RNA转染喉癌细胞Hep-2(CypA si RNA组),转染阴性对照NC si RNA的细胞(NC si RNA组)和对照组(control组)。运用real-time PCR(RT-PCR)检测CypA及CD147m RNA的表达,Western blot检测CypA、CD147的蛋白表达。MTT法检测CypA对细胞生长曲线的影响;平板克隆形成实验检测CypA在喉癌细胞增殖中的作用;Transwell法检测CypA在喉癌细胞侵袭迁移中的作用。结果:CypA si RNA转染后的喉癌细胞株Hep-2中CypA、CD147的m RNA和蛋白表达水平均明显降低(P=0.000)。CypA si RNA转染后的Hep-2细胞生长缓慢,克隆形成被抑制,control组克隆数为(235.00±23.90)、NC si RNA组为(240.67±10.07)、CypA si RNA组为(129.33±14.22)(F=40.460,P=0.000);转染后Hep-2细胞的迁移(F=497.124,P=0.000)、侵袭(F=129.787,P=0.000)能力明显降低。结论:CypA表达下调后可抑制喉癌细胞株Hep-2的增殖和侵袭迁移能力,其机制可能与抑制CD147的表达有关。展开更多
目的:探讨肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis-inducingligand,TRAIL)与化疗药物氟尿嘧啶(5-FU)、顺铂(DDP)联合使用诱导喉癌HEP-2细胞株凋亡。方法:不同浓度的TRAIL、5-FU、DDP单独及TRAIL与5-FU、...目的:探讨肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis-inducingligand,TRAIL)与化疗药物氟尿嘧啶(5-FU)、顺铂(DDP)联合使用诱导喉癌HEP-2细胞株凋亡。方法:不同浓度的TRAIL、5-FU、DDP单独及TRAIL与5-FU、DDP联合处理HEP-2细胞,用四甲基偶氮唑蓝(MTT)法检测其细胞毒作用,荧光显微镜下观察并计算凋亡率,流式细胞仪(FCM)检测细胞凋亡率。结果:HEP-2细胞对低毒性的TRAIL不敏感,对化疗药物相对敏感,TRAIL与低毒性的化疗药物联合作用于细胞后可增强杀伤效应。结论:TRAIL与低毒性的5-Fu、DDP联用表现出高效的杀灭肿瘤细胞作用。展开更多
文摘AIM: To investigate the potential roles of enhancer of zeste homolog2(EZH2), Bmi-1 and mi R-203 in cell proliferation and invasion in hepatocellular carcinoma(HCC) cell line Hep3 B.METHODS: A total of 73 patients who underwent surgical resection at Fuzong Clinical Medical College of Fujian Medical University were enrolled in this study. Hep3 B cells were cultivated in RPMI 1640 medium supplemented with 10% fetal bovine serum at 37?℃. Vectors that containing c DNA of the EZH2 gene or mi R-203 targeted sh RNA plasmid were constructed, and then transfected into Hep3 B cells. The m RNA expression of mi R-203, EZH2, and Bmi-1 was analyzed using quantitative real-time polymerase chain reaction analysis, and the protein levels of EZH2 and Bmi-1 were detected by Western blot analysis. Effect of EZH2 or mi R-203 on cell proliferation was observed by methyl thiazolyl tetrazolium assay, and cell apoptosis was assessed using flow cytometry. Besides, effect of EZH2 or mi R-203 on tumor cell invasion was detected using Transwell assay.RESULTS: The m RNA levels of EZH2 and Bmi-1 in HCC tissues and in Hep3 B cells were significantly higher compared with those in normal samples(P < 0.01), while mi R-203 level was significantly lower in HCC tissues(P < 0.01). Hep3 B cells transfected with EZH2-sh RNA or mi R-203-sh RNA showed lower expression levels of EZH2 and Bmi-1(P < 0.05). Compared with controls, Hep3 B cells transfected with EZH2-sh RNA had relative slow cell proliferation, indicating that low expression of EZH2 and Bmi-1 and overexpression of mi R-203 could inhibit Hep3 B cell proliferation(P < 0.05). The average apoptosis rate of Hep3 B cells transfected with EZH2-sh RNA vector was about 18.631%, while that of Hep3 B cells transfected with sh RNA vector was about 5.33%, suggesting that EZH2 was down-regulated by transfecting with EZH2-sh RNA, and the down-regulated EZH2 contributed to the cell apoptosis. Low expression of EZH2 and Bmi-1 and overexpression of mi R-203 could reduce Hep3 B cell invasion(P < 0.05).CONCLUSION: Our study suggests that EZH2 and Bmi-1 are up-regulated while mi R-203 is downregulated in Hep3 B cells. Mi R-203 may contribute to the metastasis and enhance apoptosis of HCC cells by regulating EZH2 and Bmi-1. Our study may provide a theoretical basis for metastasis of HCC and targeted therapy of HCC.
基金This project was supported by a grant from the Teaching and Research Award Program for Outstanding Young Teacher in Higher Education Institution of Ministry of Education of China.
文摘In order to study the effect of 5, 6-Dichloro-l-13-D-ribofuranosyl-benzimidazole （DRB） on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry （FCM） and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was （0.68±0.19）%, （1.95±0.12）%, （8.51±0.26）%, （11.26±0.17）% and （14.99±0.32）%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.
文摘目的:研究小干扰RNA(small interfering RNA,si RNA)抑制亲环素A(cyclopilin A,CypA)的表达对喉癌细胞株Hep-2增殖和侵袭迁移的影响及相关机制。方法:实验分为3组:化学合成的靶向CypA的si RNA转染喉癌细胞Hep-2(CypA si RNA组),转染阴性对照NC si RNA的细胞(NC si RNA组)和对照组(control组)。运用real-time PCR(RT-PCR)检测CypA及CD147m RNA的表达,Western blot检测CypA、CD147的蛋白表达。MTT法检测CypA对细胞生长曲线的影响;平板克隆形成实验检测CypA在喉癌细胞增殖中的作用;Transwell法检测CypA在喉癌细胞侵袭迁移中的作用。结果:CypA si RNA转染后的喉癌细胞株Hep-2中CypA、CD147的m RNA和蛋白表达水平均明显降低(P=0.000)。CypA si RNA转染后的Hep-2细胞生长缓慢,克隆形成被抑制,control组克隆数为(235.00±23.90)、NC si RNA组为(240.67±10.07)、CypA si RNA组为(129.33±14.22)(F=40.460,P=0.000);转染后Hep-2细胞的迁移(F=497.124,P=0.000)、侵袭(F=129.787,P=0.000)能力明显降低。结论:CypA表达下调后可抑制喉癌细胞株Hep-2的增殖和侵袭迁移能力,其机制可能与抑制CD147的表达有关。
文摘目的:探讨肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis-inducingligand,TRAIL)与化疗药物氟尿嘧啶(5-FU)、顺铂(DDP)联合使用诱导喉癌HEP-2细胞株凋亡。方法:不同浓度的TRAIL、5-FU、DDP单独及TRAIL与5-FU、DDP联合处理HEP-2细胞,用四甲基偶氮唑蓝(MTT)法检测其细胞毒作用,荧光显微镜下观察并计算凋亡率,流式细胞仪(FCM)检测细胞凋亡率。结果:HEP-2细胞对低毒性的TRAIL不敏感,对化疗药物相对敏感,TRAIL与低毒性的化疗药物联合作用于细胞后可增强杀伤效应。结论:TRAIL与低毒性的5-Fu、DDP联用表现出高效的杀灭肿瘤细胞作用。
