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。     

腺病毒介导NDRG2在体外对人食管鳞癌细胞系Eca-109增殖和凋亡的影响

目的:研究抑癌候选基因NDRG2在体外对人食管鳞癌细胞系Eca-109增殖和凋亡的影响.方法:腺病毒介导NDRG2转染人食管鳞癌细胞系Eca-109,逆转录酶-多聚酶链反应(RT-PCR)和Western blot法分别检测NDRG2基因的mRNA和蛋白表达水平,甲基嚷唑基四唑法(MTT法)绘制细胞生长曲线,流式细胞仪(FCM)分别分析细胞周期和细胞凋亡.结果:腺病毒介导NDRG2转染人食管鳞癌细胞系Eca-109后,NDRG2的mRNA和蛋白表达水平明显上调,细胞增殖受到明显抑制(P＜0.05);流式细胞仪检测显示,与对照组相比,感染后48小时Eca-109细胞G2期细胞明显减少(P＜0.01),S期细胞明显增多(P＜0.01);转染后的Eca-109细胞中凋亡细胞明显增多,感染48h后达16.0%.结论:过表达NDRG2可明显抑制人食管鳞癌细胞Eca-109的增殖并诱导细胞凋亡.     

腺病毒介导NDRG2在体外对人食管鳞癌细胞系Eca-109增殖和凋亡的影响

目的：研究抑癌候选基因NDRG2在体外对人食管鳞癌细胞系Eca-109增殖和凋亡的影响。方法：腺病毒介导NDRG2转染人食管鳞癌细胞系Eca-109,逆转录酶-多聚酶链反应（RT-PCR）和Western blot法分别检测NDRG2基因的mRNA和蛋白表达水平,甲基噻唑基四唑法（MTT法）绘制细胞生长曲线,流式细胞仪（FCM）分别分析细胞周期和细胞凋亡。结果：腺病毒介导NDRG2转染人食管鳞癌细胞系Eca-109后,NDRG2的mRNA和蛋白表达水平明显上调,细胞增殖受到明显抑制（P〈0.05）;流式细胞仪检测显示,与对照组相比,感染后48小时Eca-109细胞G2期细胞明显减少（P〈0.01）,S期细胞明显增多（P〈0.01）;转染后的Eca-109细胞中凋亡细胞明显增多,感染48h后达16.0%。结论：过表达NDRG2可明显抑制人食管鳞癌细胞Eca-109的增殖并诱导细胞凋亡。     

皮层肌动蛋白可通过PI3K-AKT通路调控食管鳞癌细胞的增殖和凋亡

目的探讨皮层肌动蛋白(CTTN)对食管鳞癌增殖和凋亡的影响及潜在的调控机制。方法在EC109、TE1细胞中过表达、敲低CTTN;Western blot检测PCNA、cleaved-PARP、cleaved-Caspase3以及PI3K-AKT通路的关键蛋白;CCK-8实验和平板克隆实验检测细胞活力和细胞增殖情况;软琼脂克隆形成实验评估细胞在动物体内的成瘤性,同时通过裸鼠皮下成瘤实验观察敲低CTTN对裸鼠EC109细胞皮下种植瘤生长的影响。结果成功构建CTTN稳定过表达或敲低的EC109和TE1细胞株。过表达CTTN不仅能提高PCNA、p-PI3K、p-AKT的表达水平,而且抑制cleaved-Caspase3、cleaved-PARP的表达,同时升高细胞活性,促进克隆形成(均P<0.001);敲低CTTN后PCNA、p-PI3K、p-AKT表达水平降低,cleaved-Caspase3、cleaved-PARP的表达升高,细胞活性降低,克隆形成显著减少,皮下种植瘤生长受到明显抑制(均P<0.001)。结论 CTTN通过上调PI3K-AKT通路促进食管鳞癌细胞的增殖,抑制其...     

GANT61诱导食管鳞癌细胞凋亡的作用及其机制

目的 研究Gli抑制剂GANT61诱导人食管鳞癌细胞OE21和KYSE-30凋亡的机制。方法 应用不同浓度（0、10、15 μmol/L）的GANT61处理OE21和KYSE-30细胞24 h后,Western blot观察GANT61作用于OE21和KYSE-30细胞后对Gli1、Gli2、Fas和Bcl-2蛋白表达的影响。流式细胞术观察细胞凋亡及Fas、Bcl-2蛋白的表达。结果 Western blot显示GANT61可以剂量依赖性显著下调OE21和KYSE-30细胞Gli1、Gli2和Bcl-2蛋白表达,增加Fas蛋白表达。流式细胞术检测显示不同浓度GANT61作用24 h后,OE21和KYSE-30细胞表现出不同程度的细胞凋亡并且呈药物剂量依赖性,与空白对照组相比差异均有统计学意义（P=0.000）;与空白对照组相比,OE21和KYSE-30细胞中Bcl-2蛋白表达水平显著降低且呈药物剂量依赖性,差异均有统计学意义（P=0.000）; Fas蛋白表达水平显著增加且呈药物剂量依赖性,差异均有统计学意义（P=0.000）。结论 GANT61明显诱导细胞凋亡,其作用机制可能与其通过特异性抑制OE21和KYSE-30细胞中Gli1、Gli2表达从而调节Fas和Bcl-2蛋白表达有关。     

增殖细胞核抗原和细胞凋亡表达在中晚期食管鳞癌的预后意义

探讨增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｎｇ ｃｅｌｌ ｎｕｃｌｅａｒ ａｎｔｉｇｅｎ，ＰＣＮＡ）和细胞凋亡表达中晚期食管鳞状上皮细胞癌的预后意义。方法应用微波炉加热修复抗原方法和链霉菌素－生物素免疫亲和酶技术行ＰＣＮＡ抗原染色，用末端脱氧核苷酸转移酶介导的ｄＵＴＰ缺口末端标记法进行原位凋亡染色。     

沉默GLUT-1表达对食管鳞癌细胞株TE-13增殖和迁移的影响

目的：研究沉默葡萄糖转运蛋白-1（glucose transporter protein-1,GLUT-1）的表达对食管鳞癌TE-13细胞增殖、周期及迁移的影响及可能的作用机制。方法采用慢病毒感染的方法将特异性针对GLUT-1基因的GLUT-1-shRNA转入TE-13细胞,建立稳定干扰GLUT-1表达的TE-13细胞株,分成GLUT-1-shRNA组和阴性对照组进行对比研究。采用实时荧光定量PCR和蛋白质印迹法检测GLUT-1-shRNA对TE-13细胞GLUT-1 mRNA及其蛋白表达的影响。CCK-8法、FCM法和Transwell小室迁移实验分别检测沉默GLUT-1表达后对TE-13细胞增殖、细胞周期和迁移的影响;同时采用蛋白质印迹法检测细胞迁移以及乏氧相关蛋白的表达情况。结果采用慢病毒将GLUT-1-shRNA转入TE-13细胞后,可明显下调GLUT-1蛋白（t=6.713,P<0.01）和mRNA的表达水平（t=4.181,P<0.05）。与阴性对照组相比,GLUT-1干扰后细胞的增殖至72 h（t=3.097,P<0.05）和96 h（t=3.497,P<0.05）明显被抑制,同时细胞的迁移能力可见下降,但是细胞周期至24 h未见影响;基质金属蛋白酶的MMP-9（t=6.895,P<0.01）和缺氧诱导因子HIF-1α（t=13.74,P<0.001）的表达明显下调,但是MMP-2的表达没有变化（t=2.582,P>0.05）。结论沉默GLUT-1的表达可抑制食管磷癌细胞株TE-13细胞的增殖和迁移,并明显下调细胞内MMP-9和HIF-1α蛋白的表达,为寻找靶向治疗食管癌新靶点提供了初步的实验基础。     

尼美舒利对不同COX-2表达水平的食管鳞癌细胞的抑制作用

目的：比较选择性环氧合酶-2（COX-2）抑制剂尼美舒利对不同COX-2表达水平的食管鳞癌细胞的抑制作用。方法：选取食管鳞癌细胞株EC 109、KYSE 150和TE-1,采用Western blot方法测定COX-2蛋白表达、MTT法检测细胞增殖抑制,流式细胞术检测细胞周期和细胞凋亡,观察尼美舒利对各组细胞的增殖抑制和促凋亡作用。结果：COX-2蛋白在EC 109细胞中呈高表达,KYSE 150细胞中呈中等度表达,TE-1细胞不表达COX-2蛋白。尼美舒利在50μmol/L-400μmol/L浓度区间可抑制EC 109、KYSE 150细胞的增殖（P〈0.05）,并呈剂量依赖性,在400μmol/L时对TE-1细胞有抑制作用。EC 109细胞尼美舒利的IC50值最低,KYSE150次之,TE-1最高。尼美舒利可使EC 109和KYSE 150的细胞周期阻滞于G0/G1期,并诱导细胞凋亡,但对TE-1细胞无上述作用。结论：尼美舒利对表达COX-2的食管鳞癌细胞有较好的增殖抑制和促凋亡作用。     

尼美舒利对不同COX-2表达水平的食管鳞癌细胞的抑制作用

目的:比较选择性环氧合酶-2(COX-2)抑制剂尼美舒利对不同COX-2表达水平的食管鳞癌细胞的抑制作用。方法:选取食管鳞癌细胞株EC 109、KYSE 150和TE-1,采用Western blot方法测定COX-2蛋白表达、MTT法检测细胞增殖抑制,流式细胞术检测细胞周期和细胞凋亡,观察尼美舒利对各组细胞的增殖抑制和促凋亡作用。结果:COX-2蛋白在EC 109细胞中呈高表达,KYSE 150细胞中呈中等度表达,TE-1细胞不表达COX-2蛋白。尼美舒利在50μmol/L-400μmol/L浓度区间可抑制EC 109、KYSE 150细胞的增殖(P<0.05),并呈剂量依赖性,在400μmol/L时对TE-1细胞有抑制作用。EC 109细胞尼美舒利的IC50值最低,KYSE150次之,TE-1最高。尼美舒利可使EC 109和KYSE 150的细胞周期阻滞于G0/G1期,并诱导细胞凋亡,但对TE-1细胞无上述作用。结论:尼美舒利对表达COX-2的食管鳞癌细胞有较好的增殖抑制和促凋亡作用。     

芦荟苷对食管癌细胞系KESY70增殖、凋亡和侵袭的影响

目的 探讨芦荟苷对食管癌细胞系KESY70增殖、凋亡和侵袭的影响。方法 用不同浓度的芦荟苷处理食管癌细胞系KESY70。食管癌KESY70细胞随机分为5组：未处理组（KESY70）、对照组（DMSO）和芦荟苷（10、40、80 μmol/L）组。CCK-8检测细胞活力和增殖能力。流式细胞术分析细胞凋亡,Transwell检测细胞侵袭能力,蛋白印记法分析PCNA、Cleaved caspase-3和MMP-9的表达。结果 与DMSO组相比,40、80和120 μmol/L芦荟苷处理组细胞活力明显减弱（P<0.01）。40和80μmol/L的芦荟苷处理3 d后,芦荟苷组细胞增殖能力明显降低（P<0.05）,芦荟苷组的细胞凋亡率明显高于DMSO组（P<0.05）,侵袭能力受到明显抑制（P<0.05）,PCNA、MMP-9蛋白的表达明显下降（P<0.05）,Cleaved caspase-3表达明显升高（P<0.05）。结论 芦荟苷可抑制食管癌细胞系KESY70的增殖和侵袭,促进细胞凋亡。     

AKT1 Inhibitory DNAzymes Inhibit Cell Proliferation and Migration of Thyroid Cancer Cells

AKT1 is a member of the serine/threoine AGC protein kinase family involved in thyroid cancer metabolism,growth, proliferation and survival. It is overexpressed in thyroid tumors. In this study, we designed two AKT1specific DNAzymes (DRz1 and DRz2) that target AKT1 mRNA. The results showed that DRz1 could decreasethe expression of AKT1 by 58%. Furthermore, DRz1 significantly inhibited cell proliferation, induced apoptosisand inhibited invasion in SW597 cells. In addition, down-regulation of survivin expression was associated withdecreased caspase-3, VEGF and MMP2 in SW597 cells after 24 h. In our study, the efficacy of DRz1 in decreasingAKT1 protein levels were better than DRz2. AKT1-DRz1 might have anti-tumorigenic activity and may providethe basis for a novel therapeutic intervention in thyroid cancer treatment.     

白杨素对人胃癌SGC-7901细胞增殖与凋亡的影响

 目的 探讨白杨素（Chrysin,ChR）诱导人胃癌SGC-7901细胞株凋亡的作用及机制。方法 分别用10、20、40、80μM的ChR处理人胃癌细胞株SGC-790148h,采用MTT比色法检测ChR对SGC-7901细胞的增殖抑制效应;采用丫啶橙（AO）染色、流式细胞术检测ChR诱导SGC-7901细胞凋亡的发生;应用Western-blot法检测凋亡相关基因NF-κB、Bcl-2、Bax的蛋白表达,并用计算机图像分析软件进行半定量分析。结果 MTT比色法显示的10～80μM的ChR在体外对人胃癌SGC-7901细胞有增殖抑制率为：9.71%～53.64%,该作用呈浓度依赖性;AO染色荧光显微镜观察ChR在体外能诱导人胃癌SGC-7901细胞发生凋亡,出现早期凋亡细胞及凋亡小体;流式细胞术检测结果显示：ChR呈浓度依赖性地诱导SGC-7901细胞凋亡,凋亡率为2.35%-20.8%;Western-blot检测结果显示：凋亡相关基因Bcl-2、NF-κB蛋白表达下调,Bax蛋白表达上调。结论 ChR对体外培养人胃癌细胞具有增殖抑制和诱导凋亡作用,呈浓度依赖性。ChR对人胃癌细胞增殖抑制和诱导凋亡作用机制可能与其抑制NF-κB活化、下调Bcl-2蛋白表达和上调Bax蛋白表达相关。     

CPT-11 May Provide Therapeutic Efficacy for Esophageal Squamous Cell Cancer and the Effects Correlate with the Level of DNA Topoisomerase I Protein

CPT-11 is a potent anti-cancer drug and a specific inhibitor of DNA topoisomerase I (Topo I). In this study, we aim to evaluate the effects of CPT-11 on esophageal squamous cell cancers (ESCC) and to determine the correlation between the effects and the levels of Topo I expression. We examined the growth-inhibitory effect caused by SN-38, an active metabolite of CPT-11, in 14 human ESCC cell lines established from 10 primary and 4 metastatic lesions. CPT-11 was considered effective against 5 cell lines from primary lesions and one from metastatic lesions, and thus may show therapeutic efficacy against both primary and metastatic ESCC tumors. Although Topo I mRNA levels in these 14 ESCC cell lines, as quantitated by northern blot analysis, showed no correlation with the IC₅₀ values, Topo I protein levels, as quantitated by western blot analysis, showed an inverse correlation with the IC₅₀ values. Topo I protein levels could be an indicator of sensitivity to CPT-11. We also determined Topo I protein levels in 40 ESCC tumors and matched normal mucosae. Thirty-four tumors showed 1.2-22.3-fold increases in Topo I levels. Two patients receiving pre-operative chemotherapy and one receiving radiotherapy exhibited increased Topo I protein levels in their tumor lesions. It appeared that CPT-11 could provide selective therapeutic efficacy against ESCC tumors. CPT-11 may be effective for the treatment of metastatic ESCC tumors and as a second-line anti-cancer drug for ESCC.     

miR-20 b对大肠癌细胞增殖和凋亡的影响及其机制的研究

目的：探讨miR-20b对大肠癌（ CRC）细胞增殖凋亡的影响及机制。方法将miR-20b模拟物转染CRC细胞株HCT-116,qRT-PCR检测转染后miR-20b的表达,分别应用CCK-8实验及流式细胞仪检测转染后细胞增殖和凋亡情况。 qRT-PCR和western blot检测转染后Bcl-w mRNA和蛋白的表达情况。结果转染miR-20b模拟物后,HCT-116细胞中miR-20b表达水平明显上调,HCT-116细胞增殖减弱（P＜0．05）,凋亡增强（P＜0．05）。 miR-20b可作用于Bcl-w mRNA的3′-UTR,使海肾荧光素酶活性显著降低。转染后Bcl-w mRNA及蛋白表达水平明显下调,差异均有统计学意义（P＜0．05）。结论上调HCT-116细胞中miR-20b表达水平,可抑制其靶基因Bcl-w的表达,发挥抑制细胞增殖、增加凋亡的作用。     

Marked Inhibition of Cellular Proliferation in the Normal Human Esophageal Epithelial Cells and Human Esophageal Squamous Cancer Cells in Culture by Carotenoids: Role for Prevention and Early Treatment of Esophageal Cancer

Background: Globally Esophageal cancer is a common cancer arising from human esophageal mucosal tissue.Epidemiological studies suggest inverse correlation between carotenoid intake and incident risk of this devastatingmalignancy. Methods: In an effort to examine the modulatory role of carotenoids in human esophageal carcinogenesisat a cellular level, we examined the effects of α-carotene and β-carotenes, on cell proliferation and DNA synthesis inhuman esophageal epithelial (HEE) cells and human esophageal squamous cancer (HESC) cells in in-vitro cultures.HEE and HESC cells were incubated with different concentrations of α- and β-carotenes both individually and incombination. Results: Both Carotenes significantly inhibited (p<0.05) cellular proliferation and decreased DNAsynthesis in HEE and HESC cells. The effect of α- and β-carotene together on DNA synthesis in HEE and HESC cellswas significantly greater than either carotenoid alone, suggesting a synergistic effect. Greater magnitude of cellularinhibition of DNA synthesis was observed on HEE cells than HESC cells. Conclusion: Our results suggest that acombination of α-and β-carotene may provide a novel strategy for prevention and treatment of esophageal and upperaero digestive tract cancer in humans.     

食管鳞癌中LncRNA DLEU1的表达及对食管鳞癌细胞增殖和迁移的影响

目的 研究长链非编码RNA淋巴细胞白血病缺失基因1(DLEU1)在食管鳞状细胞癌(ESCC)组织中的表达,及其对肿瘤细胞增殖和迁移的影响.方法 收集58例手术切除的ESCC及其相应癌旁组织,RT-qPCR检测ESCC组织和细胞中DLEU1的表达水平.Log rank Test法分析ESCC组织中DLEU1的表达与患者临...     

靶向Bcl-xL基因siRNA在前列腺癌细胞增殖和凋亡中的作用

目的研究Bcl-xL基因特异性RNA干扰对前列腺癌PC-3细胞株体外增殖能力和凋亡的影响。方法构建靶向Bcl-xL的小干扰RNA(siRNA)表达载体,转染PC-3细胞后,采用半定量RT-PCR和Western blot法检测转染前后Bcl-xL mRNA及蛋白表达水平的改变;采用噻唑兰(MTT)法检测细胞生长增殖情况;TUNEL试剂盒测定细胞的凋亡情况。结果靶向Bcl-xL的序列特异性的siRNA可以有效地抑制PC-3细胞Bcl-xL基因的表达。转染靶向Bcl-xL的siRNA表达质粒可以显著抑制PC-3细胞的增殖（P<0.001）,诱导细胞凋亡（P<0.001）。结论Bcl-xL基因特异性RNA干扰可以显著抑制前列腺癌PC-3细胞的体外增殖并诱导细胞凋亡。     

Silencing of Rac3 Inhibits Proliferation and Induces Apoptosis of Human Lung Cancer Cells

Background: Rac3, a member of the Rac family of small guanosine triphosphatases (GTPases), regulatesa variety of cell functions, including the organization of the cytoskeleton, cell migration, and invasion.Overexpression of Rac3 has been reported in several human cancers. However, the role of Rac3 in lung cancer(LC) has not been determined in detail. The purpose of this study was to investigate the effect of silencing of Rac3expression in human LC cells and the consequences for cell survival. Materials and Methods: Lentivirus smallhairpin RNA (shRNA) interference techniques were utilized to knock down the Rac3 gene. Gene and proteinexpression was quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.LC cell apoptosis was examined by annexin V-APC /propidium iodide staining. Results: Efficient silencing ofRac3 strongly inhibited A549 cell proliferation and colony formation ability, and significantly decreased tumorgrowth. Moreover, flow cytometry analysis showed that knockdown of Rac3 led to G2/M phase cell cycle arrestas well as an excess accumulation of cells in the G1 and S phase. Conclusions: Thus, functional analysis usingshRNAs revealed a critical role for Rac3 in the tumor growth of LC cells. shRNA silencing of Rac3 could providean effective strategy to treat LC.