TRAIL和endostatin双基因-放射治疗对人血管内皮细胞增殖、周期和凋亡的影响

目的：观察重组质粒pshuttle-Egr1-shTRAIL-shES携带的双基因TRAIL和endostatin联合X射线照射后,对血管内皮细胞ECV304增殖、周期和凋亡的影响。方法：实验分为对照组、空载体pshuttle转染组、TRAIL单基因重组质粒pshuttle-Egr1-shTRAIL转染组、endostatin单基因重组质粒pshuttle-Egr1-shES转染组和TRAIL、endostatin双基因重组质粒pshuttle-Egr1-shTRAIL-shES转染组。细胞转染采用脂质体介导的方法进行,对照组不转染。细胞转染后给予X射线照射（照射剂量分别为0、0.1、0.5、1.0、2.0和5.0 Gy）,采用ELISA法检测转染细胞中TRAIL和endostatin蛋白的表达,并分别采用MTT及PI单染或/和Annexin Ⅴ双染流式细胞术（FCM）检测TRAIL、endostatin单/双基因治疗联合放射治疗对ECV304细胞增殖、细胞周期和凋亡的影响。结果：2.0 Gy X射线照射后与0 h比较,各时间点转染pshuttle-Egr1-shTRAIL-shES的ECV304细胞上清中TRAIL和endostatin蛋白表达水平明显升高（P<0.01）,分别于12和24 h达峰值;不同剂量X射线照射可诱导TRAIL和endostatin蛋白表达,且蛋白表达水平随照射剂量的增加而明显升高（P<0.05或P<0.01）。MTT结果显示,X射线照射后, pshuttle-Egr1-shTRAIL、pshuttle-Egr1-shES和pshuttle-Egr1-shTRAIL-shES组ECV304细胞A490值均明显低于对照组和pshuttle组,并显示一定的时间-效应和剂量-效应关系,并伴有细胞凋亡率明显增加、G₂+M期细胞百分数明显上升和G₀/G₁期细胞百分数明显下降。上述细胞效应,尤以pshuttle-Egr1-shTRAIL-shES组变化最为明显,与pshuttle-Egr1-shTRAIL和pshuttle-Egr1-shES组比较差异有统计学意义（P<0.05 或 P<0.01）。结论： TRAIL和endostatin双基因联合放射治疗可抑制ECV304细胞生长,影响细胞周期进程,促进细胞凋亡,且其治疗效果优于单纯放射治疗或TRAIL/endostatin单基因-放射治疗。     

辐射诱导人TRAIL和内皮抑素双基因共表达载体的构建及鉴定

目的：构建辐射敏感Egr-1启动子诱导分泌型人肿瘤坏死因子相关凋亡诱导配体（TRAIL）和内皮抑素（endostatin）双基因共表达的载体pshuttle-Egr1-shTRAIL-shES。方法：利用聚合酶链式反应（PCR）方法从pMD19T-endostatin载体上扩增得到分泌型endostatin基因,并连接到pMD19T载体上进行测序,然后利用基因重组技术构建Egr-1启动子转录调控分泌型TRAIL和endostatin双基因表达的重组质粒pshuttle-Egr1-shTRAIL-shES。结果：PCR扩增出的650 bp片段经测序证实其序列与预期一致,说明获得的分泌型人endostatin基因正确;对pshuttle-Egr1-shTRAIL-shES及其构建过程中涉及到的多种中间载体进行PCR和酶切鉴定,结果均与预期完全一致,证实含有辐射敏感Egr-1启动子的分泌型TRAIL和endostatin双基因共表达重组质粒pshuttle-Egr1-shTRAIL-shES构建正确。结论：成功构建了辐射诱导表达的双基因共表达载体pshuttle-Egr1-shTRAIL-shES,为探讨TRAIL和endostatin双基因-放射治疗的抗肿瘤作用及其机制奠定了实验基础。     

辐射诱导表达载体pshuttle-Egr1-shTRAIL对肝癌SMMC7721细胞生长的抑制作用

目的：研究携带可溶性人肿瘤坏死因子相关凋亡诱导配体(shTRAIL)基因的辐射诱导表达载体pshuttle-Egr1-shTRAIL在人肝癌细胞株中的表达及其促凋亡作用。方法：体外通过脂质体转染SMMC7721细胞。转染细胞分别接受0(假照组)、2、5和10 Gy X线照射,ELISA法检测sTRAIL的表达;细胞分为正常SMMC7721组,分别接受0(假照组)、2和5 Gy X线照射的转染空质粒pshuttle-Egr1组及转染重组质粒pshutlle-Eyr1-shTRAIL组,采用AnnexinV-FITC凋亡检测试剂盒检测细胞凋亡率;分别取SMMC772细胞、转染STRAIL细胞、转染空载体及照射组细胞,采用克隆形成实验检测细胞存活率。结果：不同剂量X射线照射SMMC7721-shTRAIL细胞上清中可溶性TRAIL表达量明显高于假照组（P< 0.001）;与假照组比较,照射转染后SMMC7721细胞的凋亡细胞百分数明显增加（P<0.05或P<0.001）,细胞存活率明显下降（P<0.05
或P<0.001）。结论：pshuttle-Egr1-shTRAIL     

重组腺相关病毒介导内皮抑素的表达及体外活性研究

OBJECTIVE: To construct the recombinant adeno-associated viral vector containing human endostatin gene (rAAV-hEndo) and observe the biological activity of the expressed human endostatin in vitro. METHODS: rAAV-hEndo was prepared using a helper virus-free packaging system. The rAAV viral genome titer was quantified by Taqman real-time PCR, and the endostatin expressed in human umbilical vein endothelial cell line ECV304 was detected by immunofluorescence staining. The effects of endostatin on ECV340 cells were evaluated by MTT cell proliferation assay, cell cycle analysis and terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) technique. RESULTS: The viral titer of rAAV-hEndo prepared was 2 x 10(12) vg/ml and the vector had an infection efficiency of 98%. Immunofluorescence staining showed that the human endostatin protein was expressed mainly in the cytoplasm of ECV304 cells, and the proliferation of the cells was obviously inhibited by the supernatant of rAAV-hEndo, with a inhibition rate of 67.3% 72 h after the addition of the supernatant. ECV304 cells infected with rAAV-hEndo were obviously arrested in G(1) phase, and the G(1)-phase cell percentage of treatment group were significantly higher than that of control group [(72.5+/-4.0)% vs (52.1+/-2.1)%, P<0.01]. ECV304 cells infected with rAAV-hEndo demonstrated markedly enhanced apoptosis, with a significantly greater apoptotic index than that of the control cells [(32.6+/-3.2)% vs (4.2+/-1.9)%, P<0.01]. CONCLUSION: rAAV-hEndo can effectively mediate the expression of biologically active human endostatin, which may facilitate further study of antiangiogenic gene therapy with endostatin for cancers.     

重组腺相关病毒介导内皮抑素的表达及体外活性研究

目的 构建携带人内皮抑素基因的重组腺相关病毒rAAV-hEndo,介导其体外表达并检测其生物活性。方法 采用无需包装辅助病毒的双质粒共转染方法制备rAAV-hEndo,测定其滴度及感染效率。免疫荧光染色检测内皮抑素蛋白在体外感染细胞中的表达,并通过MTT法、细胞周期检测及TUNEL法检测细胞凋亡指数,观察其对人脐静脉内皮细胞ECV304的影响。结果 所制备的重组腺相关病毒滴度为2×10¹²vg/ml,感染效率达98%。免疫荧光染色显示内皮抑素蛋白主要表达于细胞胞质。rAAV-hEndo感染细胞培养上清对ECV304细胞72h增殖抑制率为67.3%。内皮细胞转染内皮抑素基因后细胞周期明显阻滞于G₁期,其G₁期占(72.5±4.0)%,与对照组(52.1±2.1)%比较有显著差异(P<0.01)。TUNEL法检测实验组内皮细胞凋亡指数为(32.6±3.2)%,与对照组(4.2±1.9)%比较有显著差异(P<0.01)。结论 所制备的重组腺相关病毒rAAV-hEndo能有效介导具有生物活性的内皮抑素表达,为进一步肿瘤抗血管生成基因治疗动物实验奠定了基础。     

Antitumor effect of betulinic acid on human acute leukemia K562 cells <Emphasis Type="Italic">in vitro</Emphasis>

The effects of betulinic acid (BA), a pentacyclic lupane-type triterpene, on the cell viability, cell cycle and apoptosis in human leukemia K562 cells were investigated. The effects of BA on the growth of K562 cells were studied by MTT assay. Apoptosis was assayed through Annexin V/propidium iodide (PI) double-labeled cytometry. The effects of BA on the cell cycle of K562 cells were studied by a PI method. The expression of Bax and capase-3 was detected by using Western blot. The results showed that BA was ...     

内皮抑素基因腺病毒载体的构建及表达

目的：构建人内皮抑素（Human Endostatin，hE）基因的腺病毒载体，并研究其对胃癌细胞株SGC-7901、MKN-45及人脐静脉内皮细胞系ECV304生物学特性的影响。方法：采用Lipofectamine2000法将含有人内皮抑素基因的质粒pCA13-hE与pBGHE3共同转染293细胞；免疫荧光法及Western Blot检测hE的表达；用不同感染复数（Muhiplicity of infection，MOI）的重组人内皮抑素基因的腺病毒感染ECV304细胞，观察细胞生长。结果：成功构建了含hE基因的腺病毒载体；经含hE基因腺病毒载体感染的人SGC-7901胃癌细胞株、MKN-45胃癌细胞株均表达hE蛋白；表达的hE蛋白具有一定的生物学活性，可以抑制人静脉内皮细胞系ECV304的生长。结论：获得了有表达功能活性的Endostatin腺病毒载体，表达产物可抑制ECV304细胞的增殖，为肿瘤的抗血管基因治疗提供了必要条件。     

KDR介导的双自杀基因重组腺病毒对ECV304细胞增殖活性、细胞周期及凋亡的影响

OBJECTIVE: To study the effect of adenovirus (Ad)-mediated fusion gene systemdriven by KDR promoter on the proliferation, apoptosis and cell cycle of human umbilical vein endothelial ECV304 cells. METHODS: The KDR-expressing ECV304 cells and LS174T cells not expressing KDR were both infected by the AdEasy-KDR-CDglyTK followed by treatment with the prodrugs 5-flurocytosine (5-FC) and/or ganciclovir (GCV) at different concentrations. The killing effects of the transfection on the cells were evaluated and bystander effects analyzed by coculturing the uninfected cells by AdKDR-CDglyTK with different ratios of infected cells. Flow cytometry was employed for determining the cell cycle distribution and electron microscopy performed to observe the pathological changes of cells. RESULTS: The infection rates of the resultant recombinant Ad (rAd) were similar in the cells and gradually increased with the increment in the multiplicity of infection (MOI) of the Ads. The infected cells exhibited different sensitivities to the two prodrugs: ECV304 cells infected with rAd were highly sensitive to the prodrugs, but the infected LS174T cells were not (P<0.001). The killing effect of CD/TK fusion gene on the target cells was much stronger than that of either single suicide gene (P<0.001), showing also obvious bystander effect. In addition, the cell cycle of ECV304 cells was arrested at S phase with morphologic features of apoptosis and necrosis as displayed by electron microscopy. CONCLUSIONS: CD/TK fusion gene system driven by KDR promoter selectively kills the KDR-CDglyTK-expressing endothelial cells, the mechanism of which may involve cell cycle arrest and necrosis and apoptosis of the cells.     

Anticancer Effect of Curcumin on Human B Cell non-Hodgkin＇s Lymphoma

Curcumin(diferuloylmethane)isayellowpig mentfromtherhizomesofturmeric(Curcumalon gaL.)andisamajorcomponentofvariousrecipesforcurry.Curcuminhasbeenreportedtohaveanumberofpharmacologicaleffectsincludinganti inflammatory,anti oxidant,antiviral,antibacterialandantitumoreffects,whichisattractingmoreandmoreattentionofinvestigators.Curcuminhasin hibitoryeffectsonmanytumorssuchasfoestom achcancer,esophagealcancer,coloncancer,livercancer,mammarytumor,bladdercancer,skincanceraswellasDMBA inducedleukem…     

Anti-cancer Effects of Deguelin on Human Leukemia K562 and K562/ADM Cells In Vitro

In order to investigate the anti-cancer effects of deguelin and on K562 and K562/ADM cells in vitro and the underlying molecular mechanism and compare the cytotoxicity of deguelin on K562, K562/ADM cells and human peripheral blood mononuclear cells (PBMCs). The effects of de- guelin on cell proliferation were assessed by MTT assay. Apoptosis were detected by AnnexinⅤ/PI double-labeled cytometry. The effects of deguelin on the cell cycle were studied by a propidium io- dide method. Our study showed that deguelin inhibited the proliferation of K562 cell and K562/ADM cell in a time- and dose-dependent manner and had minimal effects on normal human peripheral blood mononuclear cells. The ratio of IC50 value of deguelin of 24 h on K562/ADM cells to K562 cells was only 1.27, which was significantly lower than the ratio of IC50 value of ADM (higher than 20). Deguelin could induce apoptosis of K562 cells and K562/ADM cells. K562 cells were arrested at G2/M phase while K562/ADM cells were arrested at G0/G1 phase. Our results suggested that deguelin was a novel anti-leukemia agents with high efficacy and low toxicity and it is also a promising agent for reversing drug resistance.     

KDR介导的双自杀基因重组腺病毒对ECV304细胞增殖活性、细胞周期及凋亡的影响

目的 探讨腺病毒介导KDR启动子驱动的融合基因体系对人脐血管内皮细胞株ECV30的增殖活性、细胞周期及凋亡的影响.方法 以重组腺病毒AdEasy-KDR-CDglyTK体外感染表达KDR的ECV30细胞株和对照组不表达KDR的LS17T细胞株,并给予不同浓度的前药GCV(ganciclovir)和/或5-FC(5-fluorocytosine),观察该体系对ECV30细胞的杀伤效应及其旁观者效应;并以流式细胞仪检测细胞周期的变化,电镜观察细胞的病变.结果 重组腺病毒对ECV30细胞及对照组LS17T细胞的感染率相似,其感染率随腺病毒滴度的递增而增加,当MOI为200时,所有细胞株均达约100%感染.以MOI为100的重组体分别感染各细胞株表现出对前药的不同敏感性:表达KDR的ECV30细胞对前药的具有较高的敏感性,与前者相比,不表达KDR的LS17T细胞对前药不敏感(P均<0.001).融合基因的疗效优于任一单自杀基因(P均<0.001).将感染腺病毒的细胞与未感染细胞以不同混合培养,观察到该体系明显的旁观者效应.流式细胞术检测表明该体系抑制ECV30细胞DNA的合成,表现为S期细胞比率增多及G1期细胞减少(P均<0.001).同时,电镜下可见ECV30有凋亡和坏死改变.结论 KDR基因启动子可调控融合基因体系选择性杀伤人血管内皮细胞,其机制与细胞周期阻滞、凋亡及坏死有关.     

三氧化二砷对血管内皮细胞增殖和周期的影响

目的:观察As2O3对血管内皮细胞增殖、凋亡和细胞周期的干扰,探讨As2O3对血管内皮细胞生长的直接影响。方法:血管内皮细胞EVC-304体外培养,As2O3干预。采用MTT测定细胞活性,流式细胞仪测定细胞周期、细胞凋亡。结果:ECV-304细胞经过As2O3干预,其存活率明显降低,并且呈剂量依赖性。As2O3干预后,进入S期的细胞减少,细胞周期被阻在G1期,细胞可能在G1期进入凋亡。As2O3诱导早期凋亡是对照组的2.88～5.1倍,并且呈剂量依赖性;其晚期凋亡是对照组的1.17～1.67倍。结论:As2O3可直接干扰血管内皮细胞的细胞周期,阻滞细胞在G1期,并诱导细胞凋亡,进而抑制血管内皮细胞的增殖。     

Curcumin down-regulates PCNA, cyclin D1 and Bcl-XL expression in human keratinocyte cell lines

Objective：To evaluate the effects of curcumin on regulating the proliferation,cell cycle distribution,apoptosis and relevant mechanisms in keratinocyte cell lines.Methods：The human immortalized human keratinocyte lines（HaCaT cells） were treated with different doses of curcumin.The effects of curcumin on cell viability were measured by MTT assay,and the cell cycle distribution and apoptosis determined by flow cytometry.The mRNA expression changes of proliferating cell nuclear antigen（PCNA）,cyclin D1 and Bcl-xL were from real-time PCR analysis and the protein levels were detected by Western blotting.Results：Data obtained in the study showed that curcumin could cause significantly inhibitory effect on proliferation in HaCaT cells in a time- and dose-dependent manner.Cell arrest at G1/S phase and significant apoptosis were observed after being treated with curcumin for 24 h.In association with these,the expression of PCNA,cyclin D1 and Bcl-xL were decreased both at mRNA and protein levels for the same treatment.Conclusion：Curcumin can inhibit proliferation,induce cell arrest at G1/S phase and cause apoptosis in HaCaT cells.The decreased expression of PCNA,cyclin D1 and Bcl-xL induced by curcumin contributes to the above effects in vitro.     

Effects of betulinic acid on proliferation and apoptosis in Jurkat cells and its in vitro mechanism

Summary  The anti-cancer effects of betulinic acid (BA) on Jurkat cells and its in vitro mechanism were examined by using MTT assay. Apoptosis was detected by using Hoechst33258 staining and annexin-V/PI double-labeled cytometry. The effects of betulinic acid on the cell cycle of Jurkat cells were studied by propidium iodide method. RT-PCR and Western blotting were used to analyze the changes of cyclin D3, bcl-xl mRNA and protein levels in Jurkat cells after treatment with betulinic acid. Our results showed the proliferation of Jurkat cells was decreased in betulinic acid-treated group with a 24-h IC50 value being 70.00 μmol/L. Betulinic acid induced apoptosis of Jurkat cells in a time- and dose-dependent manner. The number of Jurkat cells treated with betulinic acid showed an increase in G₀/G₁ phase and decrease in S phase. After treatment with 0, 20, 60, 100 μmol/L betulinic acid for 24 h, the number of Jurkat cells was increased from (31.00±1.25)% to (58.84±0.32)% in G₀/G₁ phase, whereas it was decreased from (61.45±1.04)% to (35.82±1.95)% in S phase. PBMCs were less sensitive to the cytotoxicity of betulinic acid than Jurkat cells. The expressions of cyclin D3, bcl-xl mRNA and protein were decreased sharply in Jurkat cells treated with betulinic acid. It is concluded that betulinic acid is able to inhibit the proliferation of Jurkat cells by regulating the cell cycle, arrest cells at G₀/G₁ phase and induce the cell apoptosis. The anti-tumor effects of betulinic acid are related to the down-regulated expression of cyclin D3 and bcl-xl. Zi CHEN, Female, born in 1980, Resident This project was supported by a grant from the National Natural Sciences Foundation of China (No. 30500686).     

Effect of over-expressed LRIG3 on cell cycle and survival of glioma cells

This study examined the effects of over-expression of leucine-rich repeats and immunoglobulin-like domains 3 (LRIG3) on the cell cycle and survival of human glioma cell line U87 and U251 and explored t...     

Sulindac derivative- induced apoptosis in a human umbilical vein endothelial cell line ECV304

Objective To investigate the effects of sulindac metabolites on proliferation and apoptosis in the human umbilical vein endothelial cell line ECV304 in vitro.Methods The proliferation profile of ECV304 was determined by methyl thiazolyl tetrazolium (MTT) method. Cell cycle distribution, apoptosis and the ultrastructure of ECV304 were detected by flow cytometry (FCM) and electron microscopy, respectively.Results MTT assay showed that the sulfide inhibited the proliferation of ECV304 and its effect was dose- dependent; the IC50 was 200 μmol/L. FCM showed that the sulfide changed cell cycle distribution. The cell cycle distribution was as follows: G1 phase (control group 77. 74%±1. 58%; sulfone group 75. 63%±2. 12%; sulfide group 46. 12%±1. 60%); S phase (control group 13. 64%±1. 22%; sulfone group 16. 40±2. 30%; sulfide group 27. 26%±2. 08%); G2- M phase (control group 8. 61%±0. 67%; sulfone group 7. 98%±0. 49%; sulfide group 26. 62%±3. 54%). The apoptosis rates in the control group, sulfone group and sulfide group were 6. 08%±3. 39%, 4. 81%±2. 14% and 51. 90%±5. 67%, respectively. Sulfide reduced the proportion of G1 phase, increased the proportion of S phase, G2- M phase and the apoptosis rate significantly (P&lt;0. 01, vs control). In the sulfide- treated cells, there were nuclear fragmentation and chromosomal condensation, shrinkage of the cell and loss of contact with neighboring cells. Apoptotic bodies were observed. Sulfone showed no effect on cell proliferation, cell cycle distribution or cell morphology.Conclusions Sulfide can significantly reduce the proliferation of ECV304, change the cell cycle distribution and arrest cells in G2- M phase where apoptosis may be induced. Sulfone has no such effects on this cell line.     

miR-370-5p对前列腺癌细胞周期及增殖的影响

目的 研究miR-370-5p导入对前列腺癌细胞株DU-145和LNCaP细胞周期和增殖的影响。方法 合成miR-370-5p（实验组）和dsControl（阴性对照组）,分别转染至两个细胞株。利用Real-time PCR和Western blot法分别检测细胞转染后p21、CDK4、Cyclin D1 mRNA和蛋白的表达变化。流式细胞术分析细胞周期变化,利用MTT法和集落形成实验分析细胞活力和增殖能力。结果 Real-time PCR结果提示,转染miR-370-5p后DU-145和LNCaP细胞中p21 mRNA水平分别上调3.43倍（P<0.01）和3.06倍（P<0.01）,CDK4 mRNA水平分别下调0.51倍（P<0.01）和0.43倍（P<0.01）,Cyclin D1 mRNA水平分别下调0.31倍（P<0.01）和0.35倍（P<0.01）。Western blot法检测结果符合这一趋势。流式细胞术检测结果显示,转染miR-370-5p后,位于S期和G₂/M期的细胞比例下降,位于G₀/G₁期的细胞比例则上升,说明细胞周期被阻滞在G₀/G₁期。MTT分析结果显示,与dsControl组相比,转染miR-370-5p后,DU-145和LNCaP细胞活力明显降低。集落形成实验显示,miR-370-5p组的集落数数量明显较少,细胞增殖能力降低。结论 miR-370-5p能显著激活前列腺癌细胞中p21蛋白的表达,抑制前列腺癌细胞周期的进展和增殖。     

人Endostatin在毕赤酵母菌中的表达及其抑癌活性研究

目的在毕赤酵母中获得有活性的分泌型重组Endostatin.方法一步法抽提胎肝总RNA,RT-PCR扩增endostatin全基因.用T-A克隆技术构建克隆载体;双酶切回收550bp的endostatin基因,亚克隆入酵母表达载体.重组质粒分别以PCR、酶切及测序鉴定.电转化法将线形化的重组表达DNA转导入GS¨5感受态细胞,SDS-PAGE电泳筛选阳性重组子,G25及Heprin亲和层析柱初步纯化重组蛋白.MTT法测定重组endosatatin对血管内皮细胞增殖的抑制作用.结果RT-PCR获得550bp的特异扩增条带,T-A克隆后,PCR挑选阳性重组子,测序证实序列正确;亚克隆后的表达载体,PCR可得550bp的特异条带,EcoRI、Not I双酶切获得550bp和3kb的条带,与预期一致,DNA测序证明核苷酸序列100%正确.SDSPAGE证实重组菌较空白有一明显条带,Heparin亲和层析后,1mol/INaCl洗脱的蛋白峰在体外可特异性抑制血管内皮细胞的增殖.结论在毕赤酵母菌中成功地获得了分泌表达的活性endostatin蛋白.     

青蒿琥酯对口腔鳞癌Tca8113细胞及其移植瘤小鼠的抗肿瘤作用研究

目的 观察青蒿琥酯对口腔鳞癌Tca8113细胞增殖和凋亡的影响,以及对Tca8113细胞移植瘤小鼠肿瘤生长的影响。方法 用不同质量浓度的青蒿琥酯处理Tca8113细胞,HE染色观察细胞形态的改变;MTT法检测细胞增殖的情况;流式细胞仪检测细胞凋亡和细胞周期的变化。用口腔鳞癌Tca8113细胞建立口腔鳞癌淋巴道转移模型,ip给予移植瘤小鼠青蒿琥酯200 mg/(kg?d),连续给药10 d,观察青蒿琥酯对肿瘤生长抑制作用,以5-氟尿嘧啶（5-FU）作为阳性对照。结果 青蒿琥酯作用Tca8113细胞后,形态学观察可见细胞凋亡现象,部分细胞明显肿胀、变圆、变大,溶解成碎片,甚至死亡;细胞增殖受到抑制且呈时间、剂量相关性;青蒿琥酯能有效诱导Tca8113细胞凋亡并呈时间、剂量相关性,还能将Tca8113细胞阻滞在G₀/G₁期。体内实验显示,青蒿琥酯对Tca8113细胞移植瘤具有明显的抑制作用,抑瘤率为41.18%。结论 青蒿琥酯体内及体外对口腔鳞癌Tca8113细胞均有抑制作用,其机制可能与青蒿琥酯诱导细胞凋亡或改变细胞周期的分布有关。     

参附注射液对氧化应激血管内皮损伤及相关信号转导通路的影响

目的  研究参附注射液对氧化应激血管内皮损伤的作用及相关信号转导通路的影响。方法  培养血管内皮细胞ECV304,MTT法筛选H₂O₂造模和参附注射液的干预浓度;DCFH-DA荧光探针检测细胞内活性氧(ROS)含量;Hoechst染色和流式细胞术检测评估细胞凋亡;Western blot法检测Cleaved Caspase-3、ERK、p-ERK、Bcl-2和Bax蛋白的表达。结果  H₂O₂呈浓度依赖性诱导ECV304细胞死亡;H₂O₂促进ECV304细胞内ROS含量增加,参附注射液或N-乙酰半胱氨酸(NAC)可拮抗此过程;与对照组比较,H₂O₂能够诱导ECV304细胞凋亡,增加ERK和Caspase-3的活化,下调Bcl-2/Bax表达;与模型组比较,参附注射液或NAC能拮抗H₂O₂诱导的ECV304细胞凋亡,抑制ERK与Caspase-3的活化,上调Bcl-2/Bax表达(P < 0.05)。结论  参附注射液通过抑制ERK磷酸化和Caspase-3活化,上调Bcl-2/Bax表达,拮抗H₂O₂诱导的ECV304细胞凋亡。