芒果甙对肝癌细胞增殖的抑制和凋亡的诱导

目的 观察芒果甙对人肝癌细胞株BEL-7404的毒性和诱导凋亡作用及对细胞周期的阴滞作用，探讨芒果甙作为肿瘤化学预防药物的可能性，方法 用MTT法观察芒果甙对人肝癌细胞株增殖的抑制作用。用光学显微镜观察芒果甙对细胞的毒性作用。以流式细胞仪检测芒果甙对细胞凋亡的诱导作用和对细胞周期的干预作用。结果 不同浓度的芒果甙在不同时间对肝癌细胞株均有毒性作用，并随浓度升高和作用时间的延长而毒性作用增强，凋亡也随之增加，芒果甙阻滞肝癌细胞周期于G2/M期，20μmol/L芒果甙作用24h后上述效果开始明显。结论 芒果甙对肝癌细胞株有明显的毒性作用。能诱导肝癌细胞凋亡和阻滞细胞周期于G2/M期具有潜在药用价值。值得进一步深入研究。     

肝泰煎剂含药血清诱导人肝癌细胞系Bel-7402细胞的凋亡

目的:观察肝泰煎剂(GTJJ)含药血清诱导人肝癌细胞系Bel-7402细胞凋亡现象。方法:将GTJJ含药血清作用于人肝癌细胞系Bel-7402细胞,应用MTT检测对肝癌细胞生长抑制作用,倒置显微镜、荧光显微镜、激光共聚焦扫描显微镜等影像学方法观察细胞形态学变化,以及PI染色单染、AnnexinV-PI双染后,流式细胞术检测对细胞周期影响及诱导细胞凋亡程度。结果:MTT法检测结果显示:GTJJ含药血清具有抑制Bel-7402肿瘤细胞生长作用[其中20%GTJJ等效剂量抑制率50.09%,与生理盐水(NS)组比,P<0.01];荧光显微镜及激光共聚焦显微镜可观察到典型的凋亡形态学变化;流式细胞术检测显示,细胞周期出现G0/G1期阻滞,并出现典型的凋亡峰;AnnexinV-PI双染法检测到早期及中晚期细胞凋亡。结论:GTJJ含药血清有诱导细胞凋亡作用。     

熊果酸诱导人肝癌SMMC-7721细胞凋亡的研究

目的：观察熊果酸（UA）对人肝癌SMMC-7721细胞的增殖抑制及诱导其凋亡作用。方法：MTT法检测5、10、20、30、40、50μmol/L UA对SMMC-7721细胞生长的抑制作用,吖啶橙（AO）荧光染色、电镜和流式细胞仪检测细胞凋亡。结果：UA能显著抑制SMMC-7721细胞的增殖,其作用呈剂量依赖性。35.2μmol/L UA作用SMMC-7721细胞48小时后AO染色,荧光显微镜下可见细胞出现体积缩小,核碎裂,染色质凝集等凋亡形态改变;电镜下SMMC-7721细胞出现明显的细胞凋亡的形态学改变,细胞核染色质出现边聚和中聚,细胞内部分线粒体肿胀;SMMC-7721细胞凋亡率为（67.91±5.24）%,与对照组（2.95±0.56）%比较差异有显著性意义（P〈0.05）。结论：UA通过诱导SMMC-7721细胞凋亡抑制其生长。     

DHA对人肝癌HepG2细胞的生长抑制作用观察

体外加入二十二碳六烯酸（DHA）培养人肝癌细胞HepG2,MTT法观察细胞生长增殖情况,油红染色、荧光染色、透射电镜观察细胞形态及结构,流式细胞仪分析细胞凋亡率。结果显示,DHA能抑制HepG2生长,呈浓度和时间依赖性（P〈0．01）;细胞内有大量脂滴,并呈典型的凋亡改变,HepG2发生早期凋亡。认为DHA可抑制人肝癌细胞HepG2的生长,并使细胞发生早期凋亡。     

奥曲肽诱导人肝癌细胞凋亡的研究

应用光镜、电镜及Western blot法系统地观察了1μg/ml奥曲肽对有人肝癌细胞株SMMC－7721的形态及其内Caspase-3的表达的影响。结果发现奥曲肽能引起SMMC－7721细胞形态改变,出现核浓缩、线粒体空泡化、微绒毛减少等变化,而且能促进Caspase-3的表达并使其激活,从而诱导肝癌细胞凋亡。     

HBx 基因转染对人肝癌细胞系 HepG2细胞增殖的影响

目的探讨HBx基因转染对人肝癌细胞系Hep G2细胞增殖的影响。方法用脂质体转染法将HBx真核表达载体pc DNA3/HBx瞬时转入Hep G2细胞(转染HBx细胞组);以转染空载体细胞组和未转染Hep G2细胞组作为对照。以流式细胞仪及荧光显微镜观察各组肝癌细胞增殖情况。结果转染HBx细胞组的Hep G2肝癌细胞生长较其他两组密集。流式细胞仪检测显示转染HBx细胞组的Hep G2细胞较多处于增殖期(G2期细胞占5.68%)。荧光显微镜图像显示转染HBx细胞组处于增殖期的Hep G2细胞比率为47%,转染空载体细胞组为28%,未转染Hep G2细胞组为25%;转染HBx细胞组Hep G2细胞处于增殖期的比率与其他两组比较有统计学差异(P均<0.05)。结论 HBx基因转染能促进人肝癌细胞系Hep G2细胞增殖。     

信号分子在Genistein诱导人肝癌细胞凋亡中的作用

目的通过检测三羟异黄酮诱导肝癌细胞凋亡过程中信号分子的表达，以期探讨其诱导凋亡的分子机制。方法应用体外细胞培养技术，三羟异黄酮处理后行HE染色；采用免疫细胞化学和RT—PCR分别检测癌细胞凋亡过程中信号分子的表达情况。结果（1）三羟异黄酮作用后可见凋亡细胞，随着浓度的增加，凋亡率逐渐升高；（2）survivin蛋白的表达下调，caspase-3的蛋白表达上调；（3）survivin基因的表达逐渐降低，而caspase-3在基因水平的表达有逐渐增加趋势，但无统计学差异（P〉0．05）。结论三羟异黄酮能诱导人肝癌细胞的凋亡；在此凋亡过程中，信号分子survivin在基因与蛋白水平呈现一致性的表达降低，caspase-3在蛋白水平明显上调，但其基因表达无明显变化。推测其作用靶点可能是在转录后水平。     

苯乙酸钠对人肝癌细胞凋亡的研究

为了观察苯乙酸钠对人肝癌细胞的分化诱导作用，应用流式细胞仪（ＦＣＭ）、台盼蓝染色计数法和＾３Ｈ－ＴｄＲ掺入法观察茉乙酸钠对人肝癌细胞株的细胞周期动力学变化，另用喉癌细胞株（Ｈｅｐ－２）做对照细胞，结果显示苯乙内对人肝癌细胞呈剂量依赖性抑制，细胞周期的Ｇ１期下降，Ｓ期相对升高，但对喉癌抑制作用不明显。表明苯乙酸钠可抑制人肝癌细胞株增殖，主要是抑制细胞周期的Ｇ１期。     

中药诱导肝癌细胞凋亡的研究进展

如何诱导肿瘤细胞凋亡,以阻止肿瘤的无限生长一直是近年来肿瘤研究领域里的热点.研究表明,中药及其提取物对诱导肝癌细胞凋亡具有确切的作用,已引起学术界的普遍关注.本文将通过对近年来有关文献的回顾与复习,就中药及其提取物诱导肝癌细胞凋亡的相关研究进展作一综述.     

芍药甙对人肝癌细胞株Bel—7402增殖的影响

目的：研究芍药甙对人肝癌细胞株Bel-7402增殖的影响。方法：采用人肝癌细胞株Bcl-7402和细胞药理学方法、对比观察3种措施（阴性无药对照、阳性阿霉素对照及芍药甙）对Bel-7402细胞增殖的影响。结果：芍药甙（2mgml,1mg/ml,0.5mg/ml)对Bel-7402细胞增殖均有显著性抑制作用（均P＜0．01）,其抑制率分别为33．78％、22．22％和11．56％;增殖抑制作用随药物浓度增高而加强,呈显著的量效关系。结论：芍药甙对人肝癌细胞株Bel-7402增殖有一定的抑制作用。     

鹅脱氧胆酸衍生物HS-1200对肝肿瘤细胞株凋亡及增殖的影响

目的探讨鹅脱氧胆酸衍生物HS-1200诱导肝肿瘤细胞株凋亡及抑制增殖的作用及机制。方法分别用40、60及80p-M的HS-1200作用于肝肿瘤细胞株BEL7402,于作用后12、24及36h采用MTT检测细胞活性,流式细胞术、DNA梯状条带、荧光显微镜检测细胞凋亡,Western—blot法检测bcl-2、bax、细胞色素C及caspase-3的表达。结果HS-1200可有效地抑制BEL7402生长,其作用随药物浓度提高和作用时间延长而增强,呈一定的剂量、时间依赖性;FCM结果表明,随作用浓度和作用时间延长,凋亡率显著增加,有显著性差异（P〈0．05）;Ho-echst33258染色结果显示细胞呈凋亡形态学改变,凝胶电泳显示典型的凋亡梯形条带,Westernblot结果显示为HS-1200可提升bax、细胞色素C及caspase-3的表达,降低bcl-2的蛋白表达水平。结论HS-1200对BEL7402有显著的抑制增殖及诱导凋亡的作用,其机理可能为提升bax、细胞色素C及caspase-3的表达,降低bcl-2的表达;HS-1200可能是一种有效的治疗肝癌的化疗药物。     

奥沙利铂对人肝癌细胞株HepG2体外增殖的影响

奥沙利铂(Oxaliplatin，L-OHP)是第三代铂类化合物，其化学名为左旋反式二氨环己烷草酸铂，国际通用名为草酸铂。临床前研究表明该药对人和鼠多种肿瘤细胞均有抑制作用，且对各种顺铂耐药的肿瘤细胞株无交叉耐药。研究通过抗肿瘤药物奥沙利铂对人肝癌细胞株HepG2体外增殖的影响，为奥沙利铂应用于肝细胞癌的临床治疗提供理论依据。     

Effects of Terminalia arjuna bark extract on apoptosis of human hepatoma cell line HepG2

AIM:To investigate the effects of Terminalia arjuna(T.arjuna)extract on human hepatoma cell line(HepG2)and its possible role in induction of apoptosis.METHODS:Human hepatoma cells were treated withdifferent concentrations of ethanolic extract of T.arjunaand its cytotoxicity effect was measured by trypan blueexclusion method and lactate dehydrogenase leakageassay.Apoptosis was analyzed by light and fluorescencemicroscopic methods,and DNA fragmentation.Themechanism of apoptosis was studied with expressionof p53 and caspase-3 proteins.Glutathione(GSH)content was also measured in HepG2 cells after T.arjunatreatment.RESULTS:T.arjuna inhibited the proliferation of HepG2cells in a concentration-dependent manner.Apoptoticmorphology was observed in HepG2 cells treated with T.arjuna at the concentrations of 60 and 100 mg/L.DNAfragmentation,accumulation of p53 and cleavage ofprocaspase-3 protein were observed in HepG2 cells afterthe treatment with T.arjuna.The depletion of GSH wasobserved in HepG2 cells treated with T.arjuna.CONCLUSION:T.arjuna induced cytotoxicity in HepG2cells in vitro.Apoptosis of HepG2 cells may be due tothe DNA damage and expression of apoptotic proteins.Depletion of GSH may be involved in the induction ofapoptosis of HepG2 cells.     

Molecular mechanisms of apoptosis induced by Scorpio water extract in human hepatoma HepG2 cells

AIM: To clarify the mechanism underlying the anti-mutagenic and anti-cancer activities of Scorpio water extract (SWE). METHODS: Human hepatoma HepG2 cells were incubated with various concentrations of SWE. After 24-h incubation, cytotoxicity and apoptosis evaluations were determined by MTT and DNA fragmentation assay, respectively. After treatment with SWE, mitochondrial membrane potential (MMP) was determined by measuring the retention of the dye 3,3'-dihexyloxacarbocyanine (DiOC6(3)) and the protein expression including cytochrome C and poly-(ADPribose) polymerase (PARP) were measured by Western blotting. Caspase-3 and -9 enzyme activities were measured using specific fluorescence dyes such as Ac-DEVD-AFC and Ac-LEHD-AFC. RESULTS: We found that treatment with SWE induced apoptosis as confirmed by discontinuous DNA fragmentation in cultured human hepatoma HepG2 cells. Our investigation also showed that SWE-induced apoptosis of HepG2 cells were associated with intracellular events including disruption of MMP, increased translocation of cytochrome C from mitochondria to cytosol, activation of caspase-3, and PARP. Pre-treatment of N-acetyl-Asp-Glu-Val-Asp-CHO (Ac-DEVD-CHO), a caspase-3 specific inhibitor, or cyclosporin A (CsA), an inhibitor of MMP disruption, completely abolished SWE-induced DNA fragmentation. CONCLUSION: These results suggest that SWE possibly causes mitochondrial damage, leading to cytochrome C release into cytosol and activation of caspases resulting in PARP cleavage and execution of apoptotic cell death in HepG2 cells. These results further suggest that Scorpio may be a valuable agent of therapeutic intervention of human hepatomas.     

榄香烯体内外抑制小鼠肝癌H_（22）细胞增殖与诱导凋亡作用的研究

[目的]研究中药莪术提取物榄香烯制剂（Elemene）对小鼠肝癌H22细胞增殖的抑制及诱导凋亡的作用。[方法]体内实验,通过建立H22小鼠肝癌实体瘤模型,观察榄香烯对荷瘤小鼠肿瘤生长的抑制作用,TdT酶介导的原位缺口末端标记法（TUNEL）检测凋亡细胞。体外进行细胞培养,采用MTT法检测榄香烯对细胞增殖的影响,Annexin V-FITC/PI双染色法检测细胞的凋亡。[结果]不同实验组对荷瘤小鼠肿瘤生长的抑制作用程度不同,5-氟脲嘧啶组抑制肿瘤生长最明显,榄香烯高剂量组较低剂量组抑瘤效果明显;MTT检测到榄香烯对H22细胞株具有抑制作用,呈剂量和作用时间的依赖性;Annexin V-FITC/PI法检测到早期凋亡细胞;TUNEL法检测到凋亡细胞。[结论]榄香烯可有效抑制小鼠肝癌H22细胞的增殖,其抑制癌细胞增殖的途径是通过诱导细胞发生凋亡而完成的。     

Melatonin and Doxorubicin synergistically induce cell apoptosis in human hepatoma cell lines

AIM:To investigate whether Melatonin has synergistic effects with Doxorubicin in the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and Bel-7402.METHODS:The synergism of Melatonin and Doxorubicin inhibited the cell growth and induced cell apoptosis in human hepatoma cell lines HepG2 and Bel-7402.Cell viability was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT)assay.Cell apoptosis was evaluated using TUNEL method and flow cytometry.Apoptosis-r...     

The effect of arsenic trioxide on human hepatoma cell line BEL-7402 culturedin vitro

AIM To study the effect of a wide range of concentration of arsenic trioxide on human hepatoma cell lineBEL-7402 and its mechanism.METHODS The BEL-7402 cells were treated with arsenic trioxide (a final concentration of 0.5, 1 and2 μmol/L, respectively) in various durations or for 4 successive days. The cell growth and proliferation wereobserved by cell counting and cell-growth curve. Morphologic changes were studied under electronmicroscopy. Flow cytometry was used to assay cell-DNA distribution and the protein expression of Bcl-2 andBax was detected by immunocytochemical method.RESULTS The cell growth was significantly inhibited by the different concentrations of arsenic trioxide asrevealed by cell counting and cell-growth curve. Arsenic trioxide treatment at 0.5, 1 and 2 μmol/L, resultedin a sub-G1 cell peak. The decreased G0/G1 phase cell and the increased percentage of S phase cell were observed by flow cytometer, suggesting that the inhibiting effect of arsernic trioxide on BEL-7402 cell lay inG0/G1 phase cell. Apoptotis-related morphology, such as intact cell membrane, nucleic condensation,apoptotic body formation, can be seen under the electron microscopy. High protein expression level of Bcl-2and Bax was detected in 1 and 2 μmol/L arsenic trioxide-treated cells, but that of Bax was more significant.Arsenic trioxide treatment at 0.5 μmol/L resulted in higher expression level of Bcl-2 and lower expressionlevel of Bax compared with control (P1<0.01, P2<0.01).CONCLUSION Arsenic trioxide not only inhibited the proliferation but also induced apoptosis of humanhepatoma cell line BEL-7402. The induced-apoptosis effect of 1 and 2 μmol/L arsenic trioxide was relative tothe expression level of Bcl-2 and Bax.     

Involvement of p38 mitogen‐activated protein kinase pathway in honokiol‐induced apoptosis in a human hepatoma cell line (hepG2)

Background: Honokiol has been known to have antitumour activity. This study was conducted to evaluate the antiproliferative potential of honokiol against the hepG2 heptocellular cell line and its mechanism of action. Methods: hepG2 cells were treated with honokiol of 0–40 μg/ml concentration. The cytotoxic effect of honokiol was determined by a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. The apoptosis was evaluated by flow cytometry. Western blots were used to analyse the expression of various proteins (procaspase‐9, procaspase‐3, cleaved caspase‐3, cytochrome c, Bcl‐2, Bax, Bad, Bcl‐X_L and p38). Results: Honokiol induced apoptosis with a decreased expression of procaspase‐3 and ‐9 and an increased expression of active caspase‐3. Exposure of hepG2 cells to honokiol resulted in the downregulation of Bcl‐X_L and Bcl‐2 expression and the release of mitochondrial cytochrome c to the cytosol. In addition, honokiol activated the p38 mitogen‐activated protein kinase (MAPK) pathway, and the inhibition of this pathway by SB203580 reduced honokiol‐induced apoptosis and activation of caspase‐3. Conclusion: Honokiol induces apoptosis of hepG2 human hepatocellular carcinoma cells through activation of the p38 MAPK pathway, and, in turn, activation of caspase‐3.