Molecular Mechanism of Induction on Apoptosis of Human Esophageal Cancer HCE-4 Cells by Active Components from Astragalus membranaceus

[Objectives] To investigate the pharmacologic effects of active components from A. membranaceus on human esophageal cancer HCE-4 cells and its apoptosis mechanism. [Methods] The viabilities of HCE-4 cells were measured by MTT assay. The induction of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. [Results] Formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT,pro-caspase-3,and increase cle-caspase-3 protein expression in HCE-4 cells. [Conclusions]Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibited the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.

EXPRESSION OF PLACENTAL ALKALINE PHOSPHATASE IN ESOPHAGEAL CANCER CELL LINE Eca109

The expression and properties of alkaline phosphatase (ALP) in Eca109 cells, a cell line derived fromhuman esophageal cancer were studied with specific inhibition assay and polyacrylamide gel electrophoresis.The results showed that ALP of Eca109 cells was heat stable and was strongly inhibited by L-pheuylalanine, but slightly inhibited by urea. Preduisolone could causedramatic increase in activity of ALP, but no change in ALP isozyme and concomitant increase in lactic dehydrogenase activity were found after prednisolone treatment. The results suggested that placental alkaline phosphatase as an oncodevelopmental gene product could be expressed ectopically by Eca109 cells and prednisolone could specifically induce increase in its activity.

Incidence of human papilloma virus in esophageal squamous cell carcinoma in patients from the Lublin region

AIM:To assess the prevalence of human papilloma virus(HPV) in esophageal squamous cell carcinoma(ESCC) in the south-eastern region of Poland.METHODS:The study population consisted of 56 ESCC patients and 35 controls.The controls were patients referred to our department due to other nonesophageal and non-oncological disorders with no gross or microscopic esophageal pathology as confirmed by endoscopy and histopathology.In the ESCC patients,samples were taken from normal mucosa(56 mucosa samples) and from the tumor(56 tumor samples).Tissue samples from the controls were taken from normal mucosa of the middle esophagus(35 control samples).Quantitative determination of DNA was carried out using a spectrophotometric method.Genomic DNA was isolated using the QIAamp DNA Midi Kit.HPV infection was identified following PCR amplification of the HPV gene sequence,using primers MY09 and MY11 complementary to the genome sequence of at least 33 types of HPV.The sequencing results were computationally analyzed using the basic local alignment search tool database.RESULTS:In tumor samples,HPV DNA was identified in 28 of 56 patients(50%).High risk HPV phenotypes(16 or/and 18) were found in 5 of 56 patients(8.9%),low risk in 19 of 56 patients(33.9%) and other types of HPV(37,81,97,CP6108) in 4 of 56 patients(7.1%).In mucosa samples,HPV DNA was isolated in 21 of 56 patients(37.5%).High risk HPV DNA was confirmed in 3 of 56 patients(5.3%),low risk HPV DNA in 12 of 56 patients(21.4%),and other types of HPV in 6 of 56 patients(10.7%).In control samples,HPV DNA was identified in 4 of 35 patients(11.4%) with no high risk HPV.The occurrence of HPV in ESCC patients was significantly higher than in the controls [28 of 56(50%) vs 4 of 35(11.4%),P < 0.001].In esophageal cancer patients,both in tumor and mucosa samples,the predominant HPV phenotypes were low risk HPV,isolated 4 times more frequently than high risk phenotypes [19 of 56(33.9%) vs 5 of 56(8.9%),P < 0.001].A higher prevalence of HPV was identified in female patients(71.4% vs 46.9%).Accordingly,the high risk phenotypes were isolated more frequently in female patients and this difference reached statistical significance [3 of 7(42.9%) vs 2 of 49(4.1%),P < 0.05].Of the pathological characteristics,only an infiltrative pattern of macroscopic tumor type significantly correlated with the presence of HPV DNA in ESCC samples [20 of 27(74.1%) vs 8 of 29(27.6%) for ulcerative or protruding macroscopic type,P < 0.05].The occurrence of total HPV DNA and both HPV high or low risk phenotypes did not significantly differ with regard to particular grades of cellular differentiation,phases in depth of tumor infiltration,grades of nodal involvement and stages of tumor progression.CONCLUSION:Low risk HPV phenotypes could be one of the co-activators or/and co-carcinogens in complex,progressive,multifactorial and multistep esophageal carcinogenesis.

Human papillomavirus in esophageal squamous cell carcinoma in Colombia and Chile

AIM： To examine the presence of human papillomavirus （HPV） in esophageal squamous cell carcinoma （ESCC） specimens collected from Colombia and Chile located in the northern and southern ends of the continent, respectively.METHODS： We examined 47 and 26 formalin-fixed and paraffin-embedded ESCC specimens from Colombia and Chile, respectively. HPV was detected using GP5＋/GP6＋ primer pair for PCR, and confirmed by Southern blot analysis. Sequencing analysis of L1 region fragment was used to identify HPV genotype. In addition, P16^INK4A protein immunostaining of all the specimens was conducted.RESULTS： HPV was detected in 21 ESCC specimens （29%）. Sequencing analysis of L1 region fragment identified HPV-16 genome in 6 Colombian cases （13%） and in 5 Chilean cases （19%）. HPV-18 was detected in i0 cases （21%） in Colombia but not in any Chilean case. Since Chilean ESCC cases had a higher prevalence of HPV-16 （without statistical significance）, but a significantly lower prevalence of HPV-18 than in Colombian cases （P = 0.011） even though the two countries have similar ESCC incidence rates, the frequency of HPV-related ESCC may not be strongly affected by risk factors affecting the incidence of ESCC. HPV-16 genome was more frequently detected in p16 positive carcinomas, although the difference was not statistically significant. HPV-18 detection rate did not show any association with p16 expression. Well-differentiated tumors tended to have either HPV-16 or HPV-18 but the association was not statistically significant. HPV genotypes other than HPV-16 or 18 were not detected in either country.CONCLUSION： HPV-16 and HPV-18 genotypes can be found in ESCC specimens collected from two South American countries. Further studies on the relationship between HPV-16 presence and p16 expression in ESCC would aid understanding of the mechanism underlying the presence of HPV in ESCC.

Relationships of early esophageal cancer with human papillomavirus and alcohol metabolism

BACKGROUND It is well known that an alcohol consumption habit together with inactive heterozygous aldehyde dehydrogenase-2(ALDH2)is an important risk factor for the development of esophageal squamous cell carcinoma(ESCC).It remains controversial whether human papillomavirus(HPV)infection contributes to the occurrence/development of ESCC.There has been no study in which the relationship between ESCC and HPV in addition to alcohol dehydrogenase-1B(ADH1B)and ALDH2 genotypes was evaluated.AIM To evaluate relationships between HPV infection and development of esophageal cancer,particularly early esophageal cancer,based on ADH1B/ALDH2 polymorphisms.METHODS We conducted an exploratory retrospective study using new specimens,and we enrolled 145 patients who underwent endoscopic resection for superficial ESCC and had been observed for more than two years by both physical examination and endoscopic examination in Hokkaido University Hospital.Saliva was collected to analyze genetic polymorphisms of ADH1B/ALDH2.We performed in situ hybridization for resected specimens to detect HPV by using an HPV type 16/18 probe.RESULTS HPV was detected in 15(10.3%)of the 145 patients with ESCC.HPV-positive rates in inactive ALDH2*1/*2 and ALDH2*1/*1+*2/*2 were 10.8%and 9.8%,respectively(P=1.00).HPV-positive rates in slow-metabolizing ADH1B*1/*1 and ADH1B*1/*2+*2/*2 were 12.0%and 10.0%,respectively(P=0.72).HPV-positive rates in the heavy or moderate alcohol consumption group and the light or rare consumption group were 11.1%and 8.7%,respectively(P=0.68).HPV-positive rates in the heavy smoking group and the light or no smoking group were 11.8%and 8.3%,respectively(P=0.59).The 3-year incidence rates of secondary ESCC or head and neck cancer after initial treatment in the HPV-positive and HPVnegative groups were 14.4%and 21.4%(P=0.22),respectively.CONCLUSION In the present situation,HPV status is considered to be less important than other risk factors,such as alcohol consumption,smoking habit,ADH1B/ALDH2 polymorphisms,and HPV status would therefore have no effect on ESCC risk management.

α-细辛醚上调细胞色素C及Caspase-3表达影响食管癌Eca-109细胞的生物学行为

目的:分析α-细辛醚上调细胞色素C(cytochrome C,cytC)及Caspase-3表达对食管癌Eca-109细胞生物学行为的影响。方法:将食管癌Eca-109细胞随机等量分为4组,即低、中及高剂量(分别给予25、50及100 mg/L的α-细辛醚),另随机取等量细胞加入等体积培养液作为空白组,培养48 h后通过平板克隆实验检测4组细胞增殖情况,Annexin V/PI法检测细胞凋亡情况,RT-qPCR及Western Blot法检测细胞中cytC及Caspase-3 mRNA及蛋白表达。结果:(1)细胞克隆数在空白组最高,低剂量组显著高于高剂量组;凋亡率在空白组最低,低剂量组显著低于高剂量组。(2)4组细胞迁移至小室下的细胞数空白组>低剂量组>中剂量组>高剂量组。(3)CytC及Caspase-3蛋白在高剂量组表达显著高于空白组及低剂量组,且中剂量组表达高于空白组。(4)CytC及Caspase-3 mRNA的表达:高剂量组>中剂量组>低剂量组>空白组,差异均有统计学意义(均P<0.05)。结论:α-细辛醚抑制Eca-109细胞增殖及侵袭,并且可能通过上调cytC及Caspase-3表达促进调亡。

No evidence of HPV DNA in esophageal squamous cell carcinoma in a population of Southern Brazil

AIM:To investigate the association between human papillomavirus(HPV)and esophageal squamous cell carcinoma(ESCC)in southern Brazil.METHODS:We studied 189 esophageal samples from125 patients from three different groups:(1)102 biopsies from 51 patients with ESCC,with one sample from the tumor and another from normal esophageal mucosa distant from the tumor;(2)50 esophageal biopsies from 37 patients with a previous diagnosis of head and neck squamous cell carcinoma(HNSCC);and(3)37 biopsies from esophageal mucosa with normal appearance from 37 dyspeptic patients,not exposed to smoking or alcohol consumption.Nested-polymerase chain reaction(PCR)with the MY09/11 and GP5/6 L1primers was used to detect HPV L1 in samples fixed in formalin and stored in paraffin blocks.All PCR reactions were performed with a positive control(cervicovaginal samples),with a negative control(Human Genomic DNA)and with a blank reaction containing all reagents except DNA.We took extreme care to prevent DNA contamination in sample collection,processing,and testing.RESULTS:The histological biopsies confirmed the diagnosis of ESCC in 52 samples(51 from ESCC group and 1 from the HNSCC group)and classified as well differentiated(12/52,23.1%),moderately differentiated(27/52,51.9%)or poorly differentiated(7/52,13.5%).One hundred twenty-eight esophageal biopsies were considered normal(51 from the ESCC group,42 from the HNSCC group and 35 from dyspeptic patients).Nine had esophagitis(7 from the HNSCC and 2 from dyspeptic patients).Of a total of 189 samples,only 6 samples had insufficient material for PCR analysis:1 from mucosa distant from the tumor in a patient with ESCC,3from patients with HNSCC and 2 from patients without cancer.In 183 samples(96.8%)GAPDH,G3PDH and/orβ-globin were amplified,thus indicating the adequacy of the DNA in those samples.HPV DNA was negative in all the 183 samples tested:52 with ESCC,9 with esophagitis and 122 with normal esophageal mucosa.CONCLUSION:There was no evidence of HPV infection in different ESCC from southern Brazil.

SMAC exhibits anti-tumor effects in ECA109 cells by regulating expression of inhibitor of apoptosis protein family

BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)mimic and inhibitor of apoptosis protein(IAP)antagonist which exhibits anti-tumor effects and improves the chemical sensitivity of many cancers.AIM To ascertain the effects and mechanisms of the SMAC analog LCL161 on esophageal cancer cells.METHODS MTT assay and TUNEL assay were used to detect cell proliferation and apoptosis,respectively.Western blot analysis was used to study the molecular mechanisms of LCL161-induced death of ECA109 cells.RESULTS LCL161 decreased ECA109 cell proliferation in dose-and time-dependent manner and induced apoptosis of ECA109 cells in a dose-dependent manner.Also,LCL161 induced a significant decrease in the expression of the XIAP and significant increase in the expression of Caspase-3.In addition,Bax increased significantly with increasing concentrations of LCL161,and the relative expression of Bax was significantly different between groups.CONCLUSION These findings support the hypothesis that LCL161 can inhibit proliferation and induce apoptosis in esophageal cancer cells by regulating the expression of IAP family members,suggesting that it has potential to be an effective treatment for esophageal squamous cell carcinoma.

HPV18 E6 and E7 Intratumour Heterogeneity in Esophageal Cancer

The development of esophageal cancer accompanied by the presence of human papillomavirus (HPV) DNA into the host genome. By evaluating the expression of this virus for tumor cell origin and also their cell grows and migrations, we examined esophageal cancer clonality in the context of intra-tumor heterogeneity. In this research, we have checked the expression of HPV18 E6 and E7 in different single cell clones by the manual cell picking method in the HPV positive esophageal cancer (EC109), EC109 cell line used as a negative control, and Hela cell line used as the positive control. Quantitative real-time PCR (QRT-PCR) was run to detect the expression levels of HPV E6 and E7, Cell Counting Kit-8 (CCK-8) assay was used to examine cell proliferation, invasion assays performed using Costar chambers and wounding assay to study cell migrations in vitro. We investigated the intra-tumor heterogeneity of HPV E6 and E7 in esophageal cancer and the evaluation of the growth and migrations at the clonal level, using 10 single cell clones. In particular clones, C7 & C10 displayed a highly variable expression in both HPV E6 and E7 and weak in four clones (C1, C3, C4, and C9) consequently, the cell invasion, proliferation, and migration increase with increasing the level of HPV expression and inverse. In conclusion, the resulting based on single cell cloning showed the relationship between HPV and cell growth and migration in esophageal cancer. Future study in HPV DNA integration needed to explore the mains specific integration site of HPV DNA in esophageal cancer and molecular monitoring of the HPV for future prevention researches and also effective therapeutic strategies.

对映-贝壳杉烷型二萜化合物Pseurata H对人食道癌ECA-109细胞生长的影响

为了研究对映-贝壳杉烷型二萜化合物Pseurata H对人食道癌ECA-109细胞的增殖抑制和诱导凋亡的作用,利用倒置显微镜观察法和CCK-8检测Pseurata H对ECA-109细胞生长的影响;利用考马斯亮蓝染色法和吖啶橙/溴化乙啶(AO/EB)双染法检测药物对细胞的张力纤维损伤和细胞核形态的影响.结果显示,用浓度分别为3.125μmol/L、6.25μmol/L、12.5μmol/L、25μmol/L的Pseurata H作用ECA-109细胞24 h后,ECA-109细胞生长均受到抑制,Pseurata H对ECA-109细胞的半抑制浓度值(IC 50)为3.22μmol/L,当浓度6.25μmol/L的药物作用细胞24 h后,细胞数量减少、形态开始变圆脱落、微丝减少且核仁凝结,随着浓度增加,现象越明显.由此可见,二萜化合物Pseurata H在体外能抑制食道癌ECA-109细胞生长,具有时间-剂量依赖效应.

大蒜素对人食管癌EC-109细胞形态结构的影响

以人食管癌EC-109细胞为研究对象,探讨大蒜素对人食管癌EC-109细胞生长抑制及对形态结构的影响.采用台盼蓝拒染法测定人食管癌EC-109细胞生长抑制率,通过倒置显微镜、荧光显微镜观察不同质量分数的大蒜素作用于人食管癌EC-109细胞的形态变化.结果表明,大蒜素能抑制人食管癌EC-109细胞增殖,诱导EC-109细胞凋亡,在一定范围呈时间、剂量依赖性,作用48 h的IC50为(28±1.54)μg/mL.经大蒜素诱导后,人食管癌EC-109细胞形态结构出现典型的凋亡特征.

Marsdenia tenacissima extract induces G_0/G_1 cell cycle arrest in human esophageal carcinoma cells by inhibiting mitogen-activated protein kinase(MAPK) signaling pathway

Marsdenia tenacissima extract(MTE, trade name: Xiao-Ai-Ping injection) is an extract of a single Chinese plant medicine. It has been used for the treatment of cancer in China for decades, especially for esophageal cancer and other cancers in the digestive tract. In the present study, the potential mechanism for MTE's activity in esophageal cancer was explored. The effects of MTE on the proliferation of human esophageal cancer cells(KYSE150 and Eca-109) were investigated by the MTT assay, the Brd U(bromodeoxyuridine) incorporation immunofluorescence assay, and flow cytometric analysis. MTE inhibited cell proliferation through inducing G0/G1 cell cycle arrest in KYSE150 and Eca-109. Western blot analysis was employed to determine protein levels in the MTE treated cells. Compared with the control cells, the expression levels of the cell cycle regulatory proteins cyclin D1/D2/D3, cyclin E1, CDK2/4/6(CDK: cyclin dependent kinase), and p-Rb were decreased significantly in the cells treated with MTE at 40 mg·m L-1. In addition, MTE had an inhibitory effect on the MAPK(mitogen-activated protein kinase) signal transduction pathway, including ERK(extracellular signal-regulated kinase), JNK(c-Jun N-terminal kinase), and p38 MAPK. Moreover, MTE showed little additional effects on the regulation of cyclin D1/D3, CDK4/6, and p-Rb when the ERK pathway was already inhibited by the specific ERK inhibitor U0126. In conclusion, these data suggest that MTE inhibits human esophageal cancer cell proliferation through regulation of cell cycle regulatory proteins and the MAPK signaling pathways, which is probably mediated by the inhibition of ERK activation.

熊果酸诱导人食管癌细胞Eca-109凋亡的作用及机制

目的研究熊果酸(Ursolicacid,UA)抑制人食管癌Eca-109细胞增殖、诱导细胞凋亡的作用及机制。方法MTT比色法检测熊果酸对Eca-109细胞的增殖抑制作用;透射电镜观察经熊果酸处理后Eca-109细胞超微结构的改变;流式细胞术检测细胞周期变化及细胞凋亡率;Westernblot法检测P27kip1蛋白、凋亡相关蛋白Bcl-2、Bax的表达。结果20～50μmol/L熊果酸对Eca-109细胞具有显著的抑制作用(P<0.01);电镜下,18.32～36.65μmol/L熊果酸处理过的Eca-109细胞可见典型的凋亡形态及坏死形态;流式细胞检测显示细胞凋亡率及G0/G1期细胞随着熊果酸浓度的增加而增多,S期细胞逐渐减少;Westernblot检测结果提示熊果酸能使Bcl-2表达下降而Bax、P27kip1增加。结论熊果酸对人食管癌细胞系Eca-109具有显著的增殖抑制、诱导细胞凋亡作用。其作用与提高P27kip1蛋白的表达,将细胞阻滞在G0/G1期,下调Bcl-2的表达和增加Bax的表达有关。

尼美舒利对人食管癌细胞Eca-109 COX-2表达及生长的抑制作用

目的研究环氧化酶-2(COX-2)选择性抑制剂尼美舒利对人食管癌Eca-109细胞株COX-2表达和细胞增殖及凋亡的影响。方法MTT法测定尼美舒利对人食管癌Eca-109细胞增殖的抑制率;RT-PCR法检测Eca-109细胞COX-2mRNA表达变化;流式细胞仪检测COX-2蛋白表达、细胞周期时相分布及凋亡率的变化;光镜和琼脂糖电泳法进一步观察细胞凋亡。结果尼美舒利对人食管癌Eca-109细胞有较强的抑制作用,有明显的时间和浓度依赖性;呈浓度依赖性下调Eca-109细胞COX-2 mRNA及蛋白表达;可使Eca-109细胞G0/G1期比例增高,S期细胞减少,增殖指数降低,凋亡细胞增多。结论尼美舒利可能通过对COX-2表达的下调而诱导凋亡和细胞周期阻滞,从而抑制人食管癌Eca-109细胞生长。

槲皮素诱导人食管癌Eca-109细胞发生自噬及其作用的研究

目的明确槲皮素(quercetin,QUE)处理人食管癌Eca-109细胞后是否发生自噬及自噬在QUE抑制细胞增殖中的作用。方法用不同剂量(0、12.5、25、37.5和50μg/m L)QUE处理人食管癌Eca-109细胞24 h后,MTT检测细胞增殖率;激光共聚焦显微镜观察细胞内自噬标志蛋白LC3荧光强度;Western blot分别检测LC3和Beclin-1的表达情况。自噬抑制剂氯喹(chloroquine,CQ)预处理2 h后,以不同剂量QUE(0、12.5、25、37.5和50μg/m L)处理人食管癌Eca-109细胞24 h,MTT检测细胞增殖。结果 QUE显著抑制了Eca-109细胞的增殖,并呈剂量效应关系(P<0.05)。与对照组相比,随着QUE剂量的增加,激光共聚焦显微镜观察到在细胞内LC3荧光逐渐增强,LC3和Beclin-1蛋白表达逐渐升高。自噬抑制剂预处理后,MTT结果显示QUE+CQ组的细胞增殖率明显低于QUE单独处理组(P<0.05)。结论 QUE既可以抑制人食管癌Eca-109细胞增殖,又可以诱导细胞发生保护性自噬。抑制自噬能够增加槲皮素对肿瘤细胞的抑制作用。

D-氨基葡萄糖衍生物诱导Eca-109细胞凋亡的机制

目的探讨2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖{2-[(3-carboxy-1-oxoprogy1)amino]-2-deoxy-D-Glucose,CO-PADG}诱导Eca-109细胞凋亡的机制。方法不同浓度COPADG作用于人食管癌Eca-109细胞24h,检测Eca-109细胞的抑制率、凋亡率、细胞内活性氧(reactive oxygen spe-cies,ROS)、线粒体跨膜电位。结果Eca-109细胞凋亡率与COPADG浓度呈正相关,r=1.0,P<0.01;Eca-109细胞线粒体膜电位水平与Eca-109细胞凋亡率相关,r=1.0,P<0.01;ROS水平与Eca-109细胞凋亡率呈正相关,r=1.0,P<0.01;ROS水平与Eca-109细胞线粒体膜电位水平呈负相关,r=1.0,P<0.01。结论COPADG可促进Eca-109细胞凋亡,提高Eca-109细胞内ROS水平,并降低线粒体膜电位。实验结果提示COPADG提高ROS,降低Eca-109细胞线粒体膜电位启动细胞凋亡通路促使Eca-109细胞凋亡,并且线粒体膜电位的下降是通过提高ROS实现的。

三氧化二砷诱导人食管癌Ec109细胞凋亡伴随c-myc基因的降调节

目的 探讨三氧化二砷 (As2 O3)对食管癌 Ec10 9细胞系的生物学效应及细胞和分子机制。方法 通过四氮唑 (MTT)还原法检测三氧化二砷 (As2 O3)对食管癌 Ec10 9细胞系存活率的影响 ,从形态观察、流式细胞仪分析、DNA凝胶电泳、细胞凋亡原位检测 (TU NEL)研究三氧化二砷诱导食管癌 Ec10 9细胞凋亡的情况 ,并以 Western blot检测基因表达。结果 Ec10 9细胞经 As2 O3处理后 ,存活率明显降低。光学显微镜下可见到明显的凋亡细胞 ,在细胞周期 G1期前有低于 2倍体的凋亡峰 ;DNA凝胶电泳显示出典型的凋亡特征 :DNA有规律断裂形成的梯状图谱 ,细胞凋亡原位检测发现 DNA断裂 ,Western blot检测表明 c- myc基因的表达下降。结论 As2 O3能诱导人食管癌Ec10 9细胞凋亡 ,并伴随 c- m yc基因的降调节。

夏枯草提取物对人食管癌Eca-109细胞增殖和凋亡的影响

目的:观察夏枯草提取物对人食管癌Eca-109细胞增殖和凋亡的影响,探讨夏枯草抗肿瘤作用及机制。方法:不同浓度夏枯草作用于食管癌Eca-109细胞,利用MTT法检测细胞增殖抑制率,免疫组织化学法检测Bcl-2、Bax蛋白表达;Annexin-V-FITC/PI法检测细胞凋亡率。结果:中高剂量夏枯草抑制食管癌Eca-109细胞增殖,并呈量效和时效关系;使细胞凋亡率增高,同时Bcl-2蛋白减少,Bax蛋白表达增加。结论:夏枯草提取物可抑制食管癌Eca-109细胞增殖,使Bcl-2蛋白表达减少,Bax蛋白表达增加,细胞凋亡增加。

昼夜节律基因hPer1和hPer2对食管癌细胞株Eca-109放射敏感性的影响

目的探讨昼夜节律基因hPer1和hPer2对食管癌细胞株(Eca-109)放射敏感性的影响。方法体外培养食管癌Eca-109细胞株,通过避光和乙酰肉豆蔻佛波酯(PMA)分别诱导昼夜节律基因hPer1和hPer2节律性的表达,用RT-PCR检测出表达高峰和低谷。在hPer1和hPer2表达的高峰和低谷时,分别用6mV-X射线给予照射,通过Tunel及流式细胞术,检测肿瘤细胞的凋亡。结果昼夜节律基因表达高峰时照射与低谷时照射比较,肿瘤细胞凋亡率减少(P<0.05)。结论昼夜节律基因hPer1和hPer2高表达,能够降低食管癌细胞的放射敏感性。

人食管癌间充质干细胞对食管癌细胞株Eca-109侵袭性的影响

目的探讨人食管癌间充质干细胞(hEC-MSCs)对食管癌细胞株Eca-109侵袭性的影响。方法在体外将间质干细胞与食管癌细胞株ECA-109非接触共培养,使用RT-PCR和Western blotting的方法检测间质干细胞对ECA-109细胞株中基质金属蛋白酶-9(MMP-9)和抑癌基因E-cadherin表达的影响。结果间质干细胞可明显上调ECA-109细胞株中的MMP-9表达,显著下调ECA-109细胞株中E-cadherin的表达。结论食管癌间充质干细胞可能参与调节食管癌细胞的侵袭与转移。