Anti-hepatoma cells function of luteolin through inducing apoptosis and cell cycle arrest

The aim of this study is to explore the apoptotic induction and cell cycle arrest function of luteolin on the liver cancer cells and the related mechanism. The liver cancer cell line SMMC-7721, BEL-7402, and normal liver cells HL-7702 were treated with different concentrations of luteolin. Cell proliferation ability was tested. Morphological changes of the apoptotic cells were observed under inverted fluorescence microscope after Hoechst33342 staining. We investigated the effect of luteolin on cell cycling and apoptosis with flow cytometry. The mitochondrial membrane potential changes were analyzed after JC-1 staining. Caspases-3 and Bcl-2 family proteins expression were analyzed by real-time PCR. Cell proliferation of SMMC-7721 and BEL-7402 were inhibited by luteolin, and the inhibition was dose–time-dependent. Luteolin could arrest the cells at G1/S stage, reduce mitochondrial membrane potential, and induce higher apoptosis rate and the typical apoptotic morphological changes of the liver carcinoma cells. Q-RT-PCR results also showed that luteolin increased Bax and caspase-3 expression significantly and upregulated Bcl-2 expression in a dose-dependent manner in liver carcinoma cells. However, the normal liver cells HL-7702 was almost not affected by luteolin treatment. Luteolin can inhibit SMMC-7721 and BEL-7402 cell proliferation in a time- and dose-dependent manner. And the mechanism maybe through arresting cell cycle at phase G1/S, enhancing Bax level, reducing anti-apoptotic protein Bcl-2 level, resulting in activating caspase-3 enzyme and decrease of mitochondrial membrane potential, and finally leading to cell apoptosis.     

CoCl2诱导缺氧对Eca109细胞细胞周期及放射敏感性的影响

Objective:To investigate the change of the cell cycle,apoptosis and radiosensitivity effect by CoCl2 induced hypoxia in esophageal cancer line Eca109 cells in vitro.Methods:The hypoxia culture model induced by 150 microM CoCl2 was established.The cell cycle and apoptosis were measured with flow cytometry (FCM).The radiosensitivity was analysized with clonogenic assay after irradiation alone or combined with hypoxia in Eca109 cells in vitro.Results:Eca109 cells were treated with 150 microM CoCl2 for 24 h,cell cycle arrest in G0/G1 phase increase and decreasing arrest in S phase with longer of hypoxiac time (0-24 h),the other rate of cell cycle and apoptosis did not change obviously.The G2/M phase block was arrested obviously in radiation alone comparing with the hypoxia plus irradiated group,apoptosis did not occur in Eca109 cell line following irradiation.The DO value and cell surviving fraction of Eca109 cell was 2.48 Gy,2.44 Gy and 97.33%,96.33% in hypoxia and control group,respectively;the Dq value of Eca109 cell was 2.89 Gy,0.52 Gy,the cell surviving fraction after radiation with 4 Gy was 48.3%,21.7% in hypoxia and control group,respectively.The hypoxia decreased the radiosensitivity in esophageal cancer Eca109 cells with clonogenic assay.Conclusion:Hypoxia induced by CoCl2 influences radiosensitivity of Eca109 cell through regulating cellular proliferation rates.     

拓扑替康对食管癌细胞株Eca-109的放射增敏作用及机制

目的：研究拓扑替康（topotecan,TPT）对人食管癌细胞株Eca-109的放射增敏作用及机制。方法：MTT法检测TPT对Eca-109细胞增殖抑制作用,克隆形成实验检测TPT对Eca-109的放射增敏效应;单击多靶模型拟合细胞存活曲线并计算放射增敏比。相差显微镜观察肿瘤细胞形态学改变,流式细胞仪检测细胞周期及凋亡率。结果：TPT对Eca-109细胞有增殖抑制作用,且呈时间-剂量依赖性,10、20、40mg／L TPT的放射增敏比分别为1．22、1．29、1．36;增敏组凋亡率、G2／M期细胞比例较对照组增多（P〈0．05）。结论：TPT对食管癌细胞系Eca-109有放射增敏作用,其机制可能与促进细胞凋亡及引起G2／M期阻滞有关。     

NRAGE影响人食管癌细胞放射抗性的作用及其机制研究

背景与目的：食管癌是全球威胁人类生命和健康的常见恶性肿瘤之一，中国每年食管癌发病人数占全球发病总人数的一半以上，以食管鳞癌最为常见。放疗是食管癌三大治疗手段之一，而放射抗性是导致其治疗失败的主要原因。神经营养因子受体相互作用MAGE类药物（neurotrophin receptor-interacting MAGE homolog，NRAGE）在放射抗性细胞株TE13R120中表达量明显高于亲本TE13细胞，且NRAGE亚细胞定位变化可能参与食管癌细胞放射抗性的形成。通过基因转染构建NRAGE稳定表达的食管癌细胞系，以进一步明确NRAGE基因与食管鳞癌细胞放射抗性的关系，分析该基因影响放射抗性的具体机制。方法：通过基因转染构建NRAGE稳定表达的食管癌细胞系。采用细胞克隆形成实验检测细胞放射敏感性，采用流式细胞术检测细胞周期及凋亡；细胞划痕、Transwell侵袭实验检测细胞迁移、侵袭能力，采用实时荧光定量聚合酶链反应（real-time fluorescence quantitive polymerase chain reaction，RTFQ-PCR）和蛋白质印迹法（Western blot）检测细胞中β-catenin表达情况。组间差异采用t检验或方差分析。结果：实验分为转染过表达组（Eca109/NRAGE组）和空白对照组（Eca109组）。Eca109/NRAGE细胞中NRAGE的表达量明显高于Eca109（t=29.65，P<0.05）。照射后Eca109/NRAGE细胞的放射抗性显著高于Eca109。流式细胞术检测结果显示，Eca109/NRAGE细胞中对射线抵抗性最强的S期细胞比例增加，对射线最敏感的G ₂ /M期减少。Eca109/NRAGE细胞凋亡率较Eca109细胞降低（t=3.268，P<0.05）。Eca109/NRAGE细胞迁移和侵袭能力均高于Eca109。RTFQ-PCR和Western blot检测结果显示，β-catenin的mRNA表达及蛋白水平在Eca109/NRAGE细胞中明显高于Eca109（t=15.87，P<0.05）。结论：NRAGE参与Eca109细胞放射抗性的形成，改变细胞的细胞周期分布和凋亡情况，影响细胞迁移及侵袭能力并可能影响食管癌细胞的放射敏感性，该作用可能与激活Wnt/β-catenin信号转导通路有关。     

Effect of Lentivirus-induced shRNA Silencing CXCR4 Gene on Proliferation and Apoptosis in Human Esophageal Carcinoma Cell Line Eca109

OBJECTIVE To discuss the application of the slow virus-induced short-hairpin RNA (vshRNA) to silence the expression of CXCR4 in EsCa cell lines Eca109, and observe the effect of silencing CXCR4 on the proliferation and apoptosis of Eca109 cells in vitro. METHODS The expression plasmid of vshRNA targeting CXCR4 was constructed, with a concurrent construction of negative vshRNA expression plasmid, and without targeting any known mRNA. Real-time quantitative PCR and Western blot assay were used to determine the change of CXCR4 expression in the post-transfected EsCa cell Eca109, and MTT assay was conducted to detect the change of proliferation in EsCa Eca109 cell after silencing the CXCR4. The .ow cytometry was used to detect the change of the cell cycle and apoptosis in the post-silenced EsCa Eca109 cell in di. erent groups. RESULTS The transfection rate was respectively (87.3 ± 1.2)% and (90.1 ± 1.4)% in the CXCR4- RNAi-LV (silent group) and NC-GFP-RNAi-LV (negative control group) cellular plasmids. The vshRNA interference resulted in a down-regulation of the CXCR4 gene mRNA and protein expressions in Eca109 cells. CXCL12 promoted the proliferation of EsCa cell lines Eca109. The speed of EsCa cell proliferation became slower in the silencing group than in the normal control (also the control) and the negative control groups (P < 0.05). However, there was no significant difference in comparison of the proliferation speeds between the negative control and the normal control groups (P > 0.05). In the silencing group, the proportion of the cells in phase G0/G1, phase S and phase G2/M was respectively (69.9 ± 5.0)%, (17.1 ± 2.5)% and (13.0 ± 7.4)%, and the apoptotic rate achieved (7.27 ± 0.50)%. In the normal control group, the proportion of the cells in phase G0/G1, S and G2/M was respectively (55.9 ± 4.6)%, (30.2 ± 3.9)% and (13.8 ± 1.4)%, and the apoptotic rate was (3.30 ± 0.70)%. In the negative control group, the proportion of cells in phase G0/G1, S and G2/M was respectively (52.7 ± 7.8)%, (25.3 ± 2.3)% and (21.9 ± 7.4)%, with an apoptotic rate of (4.03 ± 1.37)%. Compared with the normal control and negative control groups, there was an apparent growth of cells in the phase G0/G1 (P < 0.05), and a greatly increased number of cells in phase S (P < 0.05) in the silencing group. There was no signi. cant di.erence in comparison of those between the normal control and negative control groups (P > 0.05). The apoptotic rate was obviously higher in the cells of the silencing group than in the normal control and the negative control groups (P < 0.05). There was no signi. cant di.erence in comparison of the apoptotic rate between the normal control and the negative control groups (P > 0.05). CONCLUSION CXCR4-vshRNA can specifically and effectively inhibit CXCR4 expression of Eca109 cells. CXCR4-vshRNA can inhibit the proliferation and enhance the apoptosis rate of Eca109 cells through intervening the expression of CXCR4, suggesting that CXCL12/CXCR4 might have an important role in the progression of Escc Thisslow virus-induced shRNA can effectively silence the expression of CXCR4 gene in the EsCa cells; block up the biological e.ect of CXCL12/CXCR4 axle; and e.ectively inhibit the potency of proliferation in the EsCa cell line Eca109, thus advancing apoptosis. It suggests that the CXCL12/CXCR4 plays an important role in the progression of EsCa.     

Comparative Studies to Evaluate Relative in vitro Potency ofLuteolin in Inducing Cell Cycle Arrest and Apoptosis in HaCaTand A375 Cells

Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology.Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanismof action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (humanimmortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin weredetermined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessedby colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis weredemonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealedthat luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with IC50 values of 37.1μM and 115.1 μM, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose andtime-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%)phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These datasuggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cellswith possible involvement of programmed cell death, providing a substantial basis for it to be developed into apotent chemopreventive template for skin cancer.     

芬太尼对食管鳞癌细胞株Eca109增殖及凋亡的影响

目的:探讨不同浓度的芬太尼对食管鳞癌细胞株Eca109增殖和凋亡的影响。方法:选用不同浓度的芬太尼（0.5、5、50、500ng/ml）干预细胞。采用四甲基偶氮唑蓝（MMT）法检测细胞增殖,流式细胞仪Annexin V/PI 双染色法测定细胞凋亡。结果:各组芬太尼孵育Eca109细胞24h后,对细胞的增殖无明显影响;孵育48h后,与对照组相比,各浓度组均能显著抑制细胞的增殖（P＜0.05）,且呈浓度和时间依赖性。芬太尼孵育Eca109细胞48h,与对照组相比,各浓度组均能诱导早期凋亡和晚期凋亡,呈浓度依赖性（P＜0.05）。结论:芬太尼可抑制食管鳞癌细胞株Eca109的增殖并诱导凋亡。     

YC-1诱导的Survivin表达抑制对食管癌胃癌细胞凋亡和增殖影响的研究

目的:观察YC-1对食管癌Eca109和胃癌SGC-7901细胞中Survivin表达的影响,探讨YC-1对食管癌和胃癌细胞生长的抑制作用.方法:Eca109和SGC-7901细胞分为对照组和YC-1实验组,细胞免疫组化和蛋白质印迹法检测YC-1对Survivin表达的影响,流式细胞仪检测YC-1对细胞凋亡和细胞周期的影响,MTT法检测YC-1对细胞增殖的影响.结果:YC-1抑制Eca109和SGC-7901细胞Sur-vivin的表达.5、10、20、30和40μmol/LYC-1对Eca109细胞48 h增殖抑制率分别为(21.95±2.05)%、(30.69±2.59)%、(50.19±2.28)%、(61.17±2.09)%和(67.50±1.99)%;对SGC-7901细胞的抑制率分别为(9.32±0.61)%、(17.86±1.33)%、(36.95±2.01)%、(47.55±1.68)%和(56.01±1.12)%.10、20和30 μmol/L YC-1作用于Eca109细胞,凋亡率分别为(14.1±0.81)%、(22.1±0.70)%和(24.6±1.01)%;作用于SGC-7901细胞,凋亡率为(15.9±0.75)%、(19.2±0.68)%、(25.4±0.95)%,P<0.05.0、20 μmol/L YC-1作用于Eca109和SGC-7901细胞,S期比例为(21.45±0.91)%,(52.19±3.53)%;(24.41±2.12)%和(54.61±4.12)%,P<0.05.结论:YC-1下调Survivin的表达,诱导食管癌Eca109和胃癌SGC-7901细胞凋亡的发生并抑制细胞增殖,有可能对食管癌和胃癌具有潜在的治疗价值.     

夏枯草对Eca109细胞的影响

目的研究夏枯草对人食管癌Eca109细胞的促凋亡作用。方法通过绘制生长曲线测定夏枯草对Eca109细胞生长增殖的影响,流式细胞术研究夏枯草诱导Eca109细胞凋亡的情况。结果100mg／mL浓度的夏枯草组细胞生长明显被抑制,Eca109细胞在100mg／mL夏枯草作用24h后,流式细胞仪测定见明显亚二倍体凋亡峰。结论100mg／mL的夏枯草可明显抑制Eca109细胞的生长并诱导其凋亡。     

半乳糖凝集素3对食管癌Eca109细胞增殖和转移的影响

目的 探讨半乳糖凝集素3（Gal3）对食管癌Eca109细胞增殖和转移的影响.方法 构建合成Gal3过表达慢病毒质粒转染食管癌Eca109细胞,应用倒置显微镜观察Gal3的过表达质粒转染食管癌细胞后荧光表达;CCK-8法检测转染前后细胞增殖能力的变化;用流式细胞术检测转染组细胞和未转染组细胞凋亡率.Transwell方法检测转染前后细胞迁移能力变化;用Western印迹检测Gal3转染前后在食管癌中表达水平变化.结果 Western印迹结果显示Gal3在食管癌细胞中表达,在Eca109/Gal3组中表达水平明显升高（t=14.33,P=0.013;t=10.28,P=0.037）.CCK-8法检测发现转染后细胞增殖能力明显升高（t=-17.277,P＜0.05;t=-13.4,P＜0.05）,流式细胞仪以Annexin-V/7-AAD双标法显示Eca109/Gal3组凋亡率明显下降（t=3.053,P＜0.05;=5.446,P＜0.05）.Transwell实验结果,Eca109/Gal3组细胞转移率高于未转染组（t=3.465,P＜0.05;t=3.252,P＜0.05）.结论 Gal3在食管癌细胞中有表达,并且其过表达能显著增强食管癌细胞的增殖、侵袭及迁移能力,明显抑制细胞的凋亡,深入研究Gal3可能为食管癌的治疗提供一种新的靶点.     

IL-27基因对Eca109细胞在裸鼠体内的成瘤抑制作用及其机制

背景与目的:细胞因子为主的肿瘤生物治疗已成为肿瘤研究领域热点之一。本研究观察白细胞介素(interleukin,IL)-27基因转染人食管癌Eca109细胞株后在裸鼠体内的成瘤抑制作用及其机制。方法:以逆转录病毒为载体,采用基因转染的方法用G418梯度筛选法建立转染IL-27基因的Eca109细胞,RT-PCR检测其基因导入情况,ELISA法检测IL-27的分泌和其诱导外周血单个核细胞(peripheral blood mononuclear cells,PBMC)产生γ-干扰素(interferon,IFN)的能力,MTT法观察Eca109/IL-27细胞生长情况。将Eca109/IL-27、Eca109/LXSN和Eca109细胞接种于裸鼠皮下,观察其成瘤性、移植瘤的生长情况并计算抑瘤率。流式细胞技术检测移植瘤组织肿瘤浸润淋巴细胞(tumor infiltrating lymphocyte,TIL)中CD16、FasL的表达和肿瘤细胞上Fas的表达。结果:RT-PCR示Eca109/IL-27细胞中有IL-27p28和IL-27EBI3亚基基因表达,Eca109/LXSN和Eca109细胞中未见表达(P<0.01),从而成功建立稳定转染的Eca109/IL-27细胞株,ELISA检测Eca109/IL-27中IL-27的分泌量高于Eca109/LXSN和Eca109细胞(P<0.01)。IL-27基因转染不影响人食管癌细胞株的体外生长(P>0.05),Eca109/IL-27诱导PBMC产生IFN-γ含量高于Eca109/LXSN和Eca109[(56.28±1.61)pg/mL vs.(12.70±0.82)pg/mL]和(11.06±0.64)pg/mL,P<0.01),Eca109/IL-27移植瘤体积较Eca109和Eca109/LXSN减小,瘤重减轻,有抑瘤作用(P<0.05);Eca109/IL-27细胞接种的肿瘤组织TIL中CD16阳性率升高,FasL的表达增高(P<0.05),肿瘤细胞表面Fas表达增加(P<0.05)。结论:IL-27基因修饰Eca109细胞在裸鼠体内产生了抑制肿瘤生长的作用,其机制可能是通过活化自然杀伤细胞,以Fas/FasL的途径产生的。     

Luteolin Suppresses Teratoma Cell Growth and Induces Cell Apoptosis via Inhibiting Bcl-2

Luteolin, which is found in plant foods, has a range of therapeutic applications. In order to examine the potential roles of luteolin in ovarian teratocarcinoma, the human ovarian teratocarcinoma cell line PA-1 was selected for functional experiments in vitro and in vivo. We demonstrated that luteolin inhibited the proliferation and colony formation of PA-1 cells in vitro. The flow cytometry results suggested that luteolin induced apoptosis of PA-1 cells in a dose-dependent manner. Immunofluorescence and qRT-PCR results showed that the expression of B-cell lymphoma-2 (Bcl-2) was decreased in luteolin-treated cells, whereas the expression of Bcl-2- associated X (Bax) was increased compared with that in the control group. In addition, luteolin inhibited the tumor growth of ovarian teratocarcinoma cells in a xenograft model. All the results suggested that luteolin induced cell apoptosis and inhibited tumor growth of PA-1 cells.     

2-（3-羧基-1-丙酰氨基）-2-脱氧-D-葡萄糖对人食管癌细胞Eta-109的致凋亡研究

目的 探讨2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖是否具有诱导人食管癌细胞Eta-109发生凋亡的作用。方法 采用体外细胞培养实验方法，应用MTT法、流式细胞仪检测、DNA琼脂糖凝胶电泳、透射电镜等技术，观察药物对细胞生长的抑制率及细胞的形态学变化，检测凋亡峰及DNALadder。结果 MTT法测得细胞抑制率具有显著的时间及浓度依赖性；透射电镜下可见到凋亡典型的形态学变化，流式细胞仪检测显示细胞周期的G期阻滞及凋亡峰；DNA琼脂糖凝胶电泳显示清晰的DNA梯状电泳。结论 2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖能够明显的抑制人食管癌细胞Eca-109的增殖，并诱导其发生凋亡。     

香加皮单体成分宝藿苷Ⅰ对食管癌细胞及裸鼠移植瘤生长抑制作用研究

 目的 研究香加皮单体成分宝藿苷Ⅰ在体内外对人食管癌细胞增殖作用的影响,以期为该成分用于临床治疗和预防食管癌提供实验依据。 方法 采用逐步分离法包括柱层析和高效液相色谱等对香加皮抗肿瘤活性成分进行分离、纯化,应用薄层层析和电喷质谱分析进行成分鉴定。采用MTT法分析不同浓度宝藿苷Ⅰ对食管癌细胞株Eca109增殖的抑制作用,应用流式细胞术分析细胞周期变化和凋亡率变化;皮下注射Eca109细胞建立裸鼠移植瘤动物模型,通过肌肉注射给药检测宝藿苷Ⅰ的体内抗肿瘤效果。 结果 宝藿苷Ⅰ可明显抑制Eca109细胞增殖（P<0.05）,并呈浓度及时间依赖性。经宝藿苷Ⅰ作用48h后,随着药物浓度的增加,Eca109 G₀/G_1期细胞明显增加（P<0.05）,S期和G2/M期细胞明显减少（P<0.05）;经宝藿苷Ⅰ作用后,Eca109细胞的凋亡率随药物作用时间的延长和宝藿苷Ⅰ浓度的增加而明显升高（P<0.01）;经宝藿苷Ⅰ（15mg/kg）治疗荷Eca109细胞裸鼠后,肿瘤生长受到明显抑制（P<0.01）,生长抑制率达（60.9±0.16）%。 结论 香加皮单体成分宝藿苷Ⅰ不仅在体外对Eca109细胞增殖具有显著的抑制作用,在体内也有较好的抗食管癌效果。其抗瘤机制可能与阻滞Eca109细胞周期发展和诱导细胞凋亡有关。     

食管鳞癌中lncRNA uc061hsf.1的表达及对Eca109、KYSE450细胞增殖、凋亡和侵袭的影响

目的:探讨uc061hsf.1基因在食管鳞癌（esophageal squamous cell carcinoma，ESCC）中表达及其对Eca109与KYSE450细胞增殖、凋亡、侵袭能力的影响。方法:本文对34例ESCC组织及癌旁组织进行RT-PCR检测，分析uc061hsf.1的表达与ESCC临床病理特征关系；通过在Eca109与KYSE450细胞系中沉默和上调uc061hsf.1表达，分别检测两组细胞增殖、凋亡、侵袭能力的变化。结果:uc061hsf.1在ESCC组织中低表达，其表达水平在分化差、淋巴结转移的患者中更低（P均＜0.05），与年龄、性别等无明显相关性；沉默uc061hsf.1可导致Eca109与KYSE450细胞系增殖及侵袭能力均明显升高，细胞凋亡无明显变化；相反，上调uc061hsf.1在这两组细胞系中的表达，则两组细胞系的增殖和侵袭能力均明显降低，且细胞凋亡率明显增加。结论:uc061hsf.1沉默可促进食管鳞癌Eca109与KYSE450细胞增殖和侵袭能力；而过表达uc061hsf.1能够抑制Eca109与KYSE450细胞增殖和侵袭，且促进细胞凋亡。     

钠尿肽受体A在食管鳞状细胞癌中的表达及对食管癌细胞迁移和侵袭能力的影响

目的 探讨钠尿肽受体A(NPRA)在食管鳞状细胞癌中的表达及其对食管癌细胞迁移和侵袭的影响.方法 采用免疫组织化学染色法检测119例食管鳞状细胞癌组织和91例癌旁组织中NPRA的表达情况,并分析食管鳞状细胞癌组织中NPRA表达情况与患者临床特征及预后的关系.采用蛋白质印迹法(Westernblot)检测食管癌细胞株Ec...     

The flavonoid Baohuoside-I inhibits cell growth and downregulates survivin and cyclin D1 expression in esophageal carcinoma via β-catenin-dependent signaling

Esophageal cancer is one of the most common malignancies and is associated with a dismal prognosis. Although treatment options have increased for some patients, overall progress has been modest. Thus, there is a great need to develop new treatments. We found that Baohuoside-I, a flavonoid extracted from a Chinese medicinal plant, exhibits anticancer activity. Here, we demonstrated that Baohuoside-I significantly inhibited Eca109 human esophageal squamous carcinoma cell proliferation and induced Eca109 cell apoptosis in vitro and in vivo. The growth inhibitory effect of Baohuoside-I on the Eca109 tumor cell line was examined by MTT assay; the induction of apoptosis was analyzed by flow cytometry. Eca109-luc cells were injected into the subcutaneous tissue of nude mice to establish xenograft tumors. Our results revealed that Baohuoside-I caused a dose- and time-dependent inhibition of cell growth and an induction of apoptosis. Furthermore, Baohuoside-I-treated cells were characterized by decreased expression of the β-catenin gene and protein in the total cell lysates. Thus, the gene and protein expression of the downstream elements survivin and cyclin D1 was downregulated. To determine the precise inhibitory mechanisms involved, further in-depth in vivo studies of Baohuoside-I are warranted. Our study provides the first evidence that Baohuoside-I inhibits tumor growth and induces apoptosis by inhibiting β-catenin-dependent signaling pathways. Thus, Baohuoside-I is a potential candidate in ESCC disease therapy.     

藤黄菌素和山萘黄素对人白血病细胞系HL-60体外增殖的抑制作用

目的观察藤黄菌素和山萘黄素对HL-60细胞体外增殖的抑制作用及其对细胞周期的影响。方法分别采用台盼蓝拒染法及流式细胞术，在不同条件下测定细胞增殖的抑制率及细胞周期的分布。结果应用台盼蓝拒染法，藤黄菌素和山萘黄素能显著抑制HL-60细胞的体外增殖并且呈现时效及量效关系。流式细胞术的结果表明，藤黄菌素和山萘黄素作用2小时可使HL-60细胞周期分别阻滞于G2／M期和G0/G1期。结论藤黄菌素和山萘黄素是两种能够明显抑制HL-60细胞体外增殖，影响其细胞周期分布的黄酮类化合物。     

外源性Egr—1基因表达对食管癌细胞株Eca109细胞生长的影响

目的：检测Egr-1蛋白在食管癌细胞株Eca109中的表达。方法：采用免疫细胞化学检测Egr-1蛋白在Eca109细胞中的表达,用脂质体基因转染法将真核表达载体PCMV－Egr-1质粒导入Eca109细胞,经Western blot和免疫细胞化学检测Egr-1蛋白在转染的Eca109细胞中呈高表达。结果Eca109细胞的Egr-1蛋白表达阴性,当导入外源性Egr-1基因后,Egr-1捕达阳性,表达Egr-1蛋白的Eca109的细胞形态上发生明显的的变化,表现为细胞变偏,呈散在生长,结论Egr-1蛋白的表达在一定程度上能抑制Eca109细胞生长。