Effect of Octadecadienoic Acid on Proliferation and Apoptosis of Glioma Cells and Its Mechanism

[Objectives] To explore the inhibitory effect of octadecadienoic acid (ODA) on proliferation and apoptosis of glioma cells and its mechanism. [Methods] Cultured human glioma cells (cell density 2×10^(6) cells/L) were divided into three groups: solvent control group (DMSO, 30 μL/L), 5-FU group (10 mg/L) and octadecadienoic acid group (0.3, 0.6, 1.2 mg/L). The toxic effects of ODA on glioma cells were detected by trypan blue and thiazolium blue (MTT). The expression of P53, PI3K, P21, PKB/Akt and caspase-9 protein in glioma cells were detected by enzyme-linked immunosorbent assay (ELISA). [Results] The cell count under optical microscope showed that the inhibition rate of cell proliferation in low, medium and high dose ODA groups and 5-FU group was significantly higher than that in solvent control group ( P <0.01), but there was no significant difference compared with 5-FU group ( P >0.05). The results of MTT showed that compared with the solvent control group, the inhibition rate of cell proliferation in low, medium and high dose ODA groups and 5-FU group significantly increased ( P <0.01);compared with 5-FU group, the inhibition rate of cell proliferation in high dose ODA group significantly increased ( P <0.01). The results of flow cytometry showed that compared with the solvent control group, the number of cells in G_(0)/G_(1) phase increased significantly ( P <0.05, P <0.01), the number of cells in G_(2)/M phase decreased significantly ( P <0.01) and the apoptosis rate increased significantly ( P <0.01) in the low, medium and high dose ODA groups and 5-FU group;compared with 5-FU group, the number of cells in G_(2)/M phase decreased significantly ( P <0.01) and the apoptosis rate increased significantly ( P <0.01) in ODA group. ELISA testing results showed that the expression levels of P53, P13K and PKB/Akt in low, medium and high dose ODA groups and 5-FU group were significantly lower than those in solvent control group ( P <0.01), and only the expression level of protein in high dose ODA group was significantly lower than that in 5-FU group ( P <0.01);the expression levels of P21 and caspase-9 in low, medium and high dose ODA groups and 5-FU group were significantly higher than those in solvent control group ( P <0.05, P <0.01), but the expression level of protein in high dose ODA group was significantly higher than that in 5-FU group ( P <0.01). [Conclusions] ODA can obviously inhibit the proliferation of glioma cells and induce apoptosis. The mechanism is related to up-regulation of P21, caspase-9, down-regulation of P53, PI3K, PKB/Akt, inhibition of cell division cycle and decrease of PI3K-Akt signal transduction pathway.

MEK/ERK signaling pathway in apoptosis of SW620 cell line and inhibition effect of resveratrol

Objective:To study the involvement of MAPK MEK/ERK signaling transduction pathway in the apoptosis process of SW620 tumor cell line and the inhibition effect of resveratrol.Methods:SW620 cell lines were divided into 5 groups,namely,control group.PD98059 group,low-dose resveratrol group,mid-dose resveratrol group and high-dose resveratrol group.The inhibition rate of cell proliferation was detected by MTT method.The expression of apoptotic molecules and MEK/ERK signaling pathway related proteins were assayed by realtime PCR and Western blotting.Results:Compared with control group,the proliferation of cells treated with resveratrol was significantly inhibited.In the case of apoptotic molecules,the expression of Bax,Caspase 3 and Caspase 9 was increased significantly while the expression of anti-apoptotic molecule Bcl2 was decreased significantly in resveratrol groups with a dosedependent manner.In the case of molecules in MEK/ERK signaling pathway,the expression of Ras,Raf,MEK and ERKl/2 was decreased significantly in resveratrol groups with a dose-dependent manner.Conclusions:PD98059 and resveratrol can effectively inhibit the proliferation of SW620 through inhibiting the MEK/ERK signaling pathway.

Inhibitive effects of anti-oxidative vitamins on mannitol-induced apoptosis of vascular endothelial cells

Objective: Study blood vessel injury and gene expression indicating vascular endothelial cell apoptosis induced by mannitol with and without administration of anti-oxidative vitamins. Methods: Healthy rabbits were randomly divided into four groups. Mannitol was injected into the vein of the rabbit ear in each animal. Pre-treatment prior to mannitol injection was per- formed with normal saline (group B), vitamin C (group C) and vitamin E (group D). Blood vessel injury was assessed under electron and light microscopy. In a second experiment, cell culture specimen of human umbilical vein endothelial cells were treated with mannitol. Pre-treatment was done with normal saline (sample B), vitamin C (sample C) and vitamin E (sample D). Total RNA was extracted with the original single step procedure, followed by hybridisation and analysis of gene expression. Results: In the animal experiment, serious blood vessel injury was seen in group A and group B. Group D showed light injury only, and normal tissue without pathological changes was seen in group C. Of all 330 apoptosis-related genes analysed in human cell culture specimen, no significant difference was seen after pre-treatment with normal saline, compared with the gene chip without pre-treatment. On the gene chip pre-treated with vitamin C, 45 apoptosis genes were down-regulated and 34 anti-apoptosis genes were up-regulated. Pre-treatment with vitamin E resulted in the down-regulation of 3 apoptosis genes. Conclusion: Vitamin C can protect vascular endothelial cells from mannitol-induced injury.

Inhibitory effects and mechanism of rhBMP-2 on the proliferation of SHG44 human glioma cells

Objective To investigate the inhibitory effects of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the proliferation of human glioma cells (SHG44) in vivo and in vitro, and analyze its mechanism. Methds: Human SHG44 glioma cells were cultured in vitro. The growth curve of SHG44 cells with or without rhBMP-2 was ploted. The pro- liferative action of rhBMP-2 on SHG44 cells was determined by MTT method The cell cycle, ultrastructure and DNA frag-ments of SHG44 cells were detected by now cytometry (FCM), electron microscopy and agarose gel electrophoresis agarose respectively. The growth state of SHG44 glioma under the skin of nude mice was observed with the local injection of rhBMP-2. Results:rhBMP-2 could inhibit the growh of SHG44 cells in vitro. The DNA content of SHG44 cells increased in G1 phase, and decreased in S phase obviously (p<0.01) .An apoptosis spike (12.3%) was demonstrated by FCM as- say. The nucleus chromatin condensation, margination,breakage and apoptosis cells were observed under the electron mi- croscopy.The DNA electrophoresis showed a ladder band (DNA ladder). SHG44 glioma growth in mice was relatively slow-~ er and its volume was smaller with local injection of rhBMP-2 than. those without rhBMP-2 injection (P < 0.01). Conclu-sion: SHG44 cells could be arrested at G1/S phase and the cell growht could be significantly suppressed by rhBMP-2. The process might be carried out thsough rhBMP-2-induced apoptosis in human glioma cells.

Effect of trkA kinase inhibitor on apoptosis of human pterygium fibroblasts

Objective： The aim of this study was to investigate the growth inhibition effects and the mechanisms of trkA kinase inhibitor on human pterygium fibroblasts（HPF）. Methods： Three normal conjunctivas and eleven pterygiums were surgically collected, trkA and p75 was examined in different tissues by immunohistochemistry, trkA and p75 expression was detected in HPF by immumofluorescence method. After being treated with 10-80 nmol/L trkA kinase inhibitor for 24-96 h, the growth activities of HPF were studied by MTT colorimetry. Caspase-3 was inspected in HPF by spectrophotometric method. The apoptosis of HPF was detected by flow cytometry. Results： Expression of trkA and p75 was significantly observed in epithelium and fibroblast of pterygium tissues. Expression of trkAwas observed in epithelium and fibroblast of normal conjunc- tiva. Expression of p75 was only observed in epithelium of normal conjuncUva, trkA kinase inhibitor could effectively inhibit the growth of human pterygium fibroblasts in vitro in time and dose dependent manners. Caspase-3 expression was increased. The rates of apoptosis were 8.26%-29.62% （P 〈 0.01）. Conclusion： TrkA and p75 possibly participate in genesis and development of pterygium, trkA kinase inhibitor could induce apoptosis of human pterygium fibroblasts. Inducing apoptosis through changing the ratio of trkA and p75 in fibroblasts and up-regulate Caspase-3 was probably one of its molecular mechanisms.

Trojan horses in tumor:engineered pyroelectric and photodynamic nanocomposites for NIR-induced cell apoptosis and tumor growth inhibition

It is desirable but always challenging to develop a cutting-edge tumor treatment strategy with high therapeutic efficacy,lesiontargeted precision and mild accessibility.Compared to traditional treatment modalities,photodynamic therapy has been widely studied since the generation of reactive oxygen species(ROS)at cancerous lesions unprecedentedly offers a convenient approach for localized tumor eliminations.Nevertheless,the consumption of oxygen for ROS production in a hypoxic tumor microenvironment has dramatically limited its feasibility and efficacy.Herein,the engineered nanocomposites of BTO@PDA-ICGHA with photodynamic and pyroelectric performances have been fabricated and applied to the photodynamic-pyroelectric dynamic treatments.The continuing ROS production derived from intracellular oxygen(O_(2))and water(H_(2)O)by laser irradiation contributed to the superb tumor cell apoptosis and significant tumor growth inhibition.Thus,this study has validated a new concept by depositing the engineered nanocomposites at the tumor just like Trojan horses,facilitating ROS release as killers and exerting the NIR-induced cell apoptosis and tumor growth inhibition with high therapeutic efficiency and expectable translational perspectives.

Proliferation-Inhibiting and Apoptosis-lnducing Effects of Ursolic Acid and Oleanolic Acid on Multi-Drug Resistance Cancer Cells in Vitro

Objective：To investigate the proliferation-inhibiting and apoptosis-inducing effects of ursolic acid（UA） and oleanolic acid（OA） on multi-drug resistance（MDR） cancer cells in vitro.Methods：UA and OA in different concentrations（0-100μmol/L） were added separately to cultures of different cancer cell lines, including the human colon cancer cell lines SW480 and SW620,human acute myelocytic leukemia cancer cell lines HL60 and HL60/ADR,human chronic myelogenous leukemia cell lines K562 and K562/ADR,and the human breast cancer cell lines MCF-7 and MCF-7/ADR.Effects of UA and OA on cell proliferation were detected by 3-（4,5-dimethyl-2-thiazole）-2-5-biphenly-tetrazole bromide（MTT） method and effects on cell apoptosis were tested by flow cytometry（FCM） and Western blot at 24,48,and 72 h after treatment.Results：Both UA and OA showed significant inhibition on parent and MDR cell lines in a time- and concentration-dependent manner;the drug-resistant multiple of them on K562 and K562/ADR as well as on HL60 and HL60/ADR was 1;the effects of UA were better than those of OA in inhibiting cell growth of solid colonic cancer and breast cancer.After SW480 cells were treated by UA at the concentrations of 0-40μmol/L for 48 h,FCM showed that annexin V （AV） positive cells and hypodiploid peak ratio increased along with the increase in the drug＇s concentrations; and Western blot found that expressions of Bcl-2,Bcl-xL and survivin decreased in a concentration-dependent manner.Conclusions：Both UA and OA have antitumor effects on cancer cells with MDR,and the optimal effect is shown by UA on colonic cancer cells.Also,UA shows cell apoptosis-inducing effect on SW480,possibly by way of down-regulating the expressions of apoptosis antagonistic proteins,Bcl-2,Bcl-xL,and survivin.

Identifying Combinatorial Growth Inhibitory Effects of Various Plant Extracts on Leukemia Cells through Systematic Experimental Design

Plant extracts are widely studied for their anti-cancer and cancer preventive effects. In this study, we compared the leukemia growth inhibition effects of seven different plant extracts, theaflavin, epigallocatechin gallate (EGCG), epicathechin (EC), apigenin, quercetin, chrysin and tannic acid, in vitro using the K562 erythroleukemia cell line and application of the design of experiments (DoE) methodology. Our systematic approach enabled us to isolate the main factor contribution, two-factor interactions and produced interaction relationships and/or models to describe growth inhibitory effects of different plant extracts when they are used in combination. The results identified tannic acid as the most significant inhibitor in this group and had synergistic effects with EGCG at specific concentrations. The fitted model of their combined effects showed that the most potent combination is at low concentrations of tannic acid (10 - 20 μM) and high concentrations of EGCG (80 - 100 μM). We further showed that tannic acid induced both growth inhibition and apoptosis in K562 cells in ranges between 10 - 100 μM. The polyphenol caused cell cycle arrest at G2- phase under the higher concentrations. In summary, use of DoE techniques effectively identified the most prominent inducer in this group of plant bioactive compounds and produced combinatorial bioactivity of various polyphenols and flavonoids over the entire range of concentrations under study. This study exemplifies the usefulness of DoE and serves as a guide in its utility for in vitro assessment of bioactivity in plant constituents.

Role of Extracelluar Regulated Protein Kinases in FTY720-induced Apoptosis of Leukemia Cell Lines HL-60 and U937

The effects of a novel immunosuppressive agent FTY720 on proliferation inhibition and apoptosis of acute leukemia cell lines HL 60 and U937, and the role of extracelluar regulated protein kinase (ERK) in the course of proliferation inhibition and apoptosis induced by FTY720 were studied. The proliferation inhibition rate of HL 60 and U937 cells by various concentrations of FTY720 was detected by MTT assay. Cell apoptosis was detected by DNA fragment analysis and flow cytometry. The phosphorylated ERK1/2 protein expression was observed by Western blotting. The change of intracellular distribution of ERK1/2 protein was identified by SP immunohistochemical staining. The results showed that FTY720 could inhibit the growth of HL 60 and U937 cells effectively in a dose dependent manner. After incubation with FTY720 for 24 h, apoptosis was observed in HL 60 and U937 cells. The intracellular expression of phosphorylated ERK1/2 protein was also down regulated and the distribution of ERK1/2 protein in cell nuclear was reduced during FTY720 induced apoptosis. So, that FTY720 inhibited ERK1/2 phosphorylation might mediate the role of FTY720 induced apoptosis and proliferation inhibition of leukemia cells.

Xiaoji Decoction(消积饮) Inhibited Cell Proliferation and Induced Apoptosis through Akt Signaling Pathway in Human Lung Cancer A549 Cells

Objective： To investigate the inhibitive effect and the underlying mechanism of Xiaoji Decoction （消极饮 XJD） in human lung cancer A549 cells. Methods： A549 cells in logarithmic proliferation were cultivated in RPMI-1640 containing 10% low, medium or high dosages of XJD serum. The inhibitive effect of XJD in A549 cell proliferation was assessed by methylthiazolyldiphenyl-tetrazolium bromide （MTT） assay. The pro-apoptotic effect of XJD in A549 cells was observed by fluorescence microscope via Hoechst 33258 staining. The role of the Akt signaling pathway was observed by examining the presence of p-Akt protein by Western blot and the mRNA expression of downstream proteins such as Bcl-2/BcI-XL-associated death promoter （BAD） and caspase-9 by real time polymerase chain reaction. Results： MTT assay revealed that XJD could inhibit A549 proliferation in a dose- and time-dependent manner. Hoechst 33258 staining showed that XJD induced the typical nuclear apoptotic morphology after XJD treatment. Moreover, XJD could reduce the phosphorylation of Akt and increase the mRNA expression of BAD and caspase-9. Conclusions： XJD can inhibit the proliferation of A549 cells in a dose- and time-dependent manner through signaling Akt pathway via up-regulating the expression of BAD and caspase-9. XJD may provide a novel therapeutic model for lung cancer and deserve further study.

Effects of Radix Salviae Miltiorrhizae on Proliferation,Apoptosis and c-myc Protein Expression of Fibroblast in Culture of Kindney with Lupus Nephritis

Objectiv:To observe the effects ofRadix Salviae Miltiorrhizae (SM) on humanfibroblast in culture of kidney with lupus nephritis (LN ). Methods: Fibroblasts wereisolated from culture of kidney biopsy of LN patients, and effect of SM on 3H-TdR incorporated rate of fibroblasts was observed. Theapoptosis and c-myc expression were detectedin the same time by flow cytometry.Results:SM could inhibit the proliferation of fibrolast,and promote the programmed cell deaththrough upregulate c-myc protein expression inhuman renal fibroblasts. Conclusions: Longterm administration of SM in large dosagecould be effective on interstial fibrosis of LN,so that to prevent or reduce the scar tissue for-mation and teatrd the occurrence of uremia.

斑蝥酸钠在诱导人宫颈癌Hela细胞凋亡相关基因表达中的作用机制研究

目的:探讨斑蝥酸钠在诱导人宫颈癌Hela细胞凋亡相关基因表达中的作用机制。方法:2023年1月—2023年12月购买1株人宫颈癌Hela细胞,培养至对数生长期后开始实验,将其分为4组,分别加斑蝥酸钠0μmol/L(空白对照组)、4μmol/L(4μmol/L组)、8μmol/L(8μmol/L组)和16μmol/L(16μmol/L组),每个浓度分别设置5个复孔(每组样本量为5),以细胞活力试验(CCK-8)检测细胞增殖情况,以流式细胞仪检测细胞凋亡情况,以实时荧光定量多聚核苷酸链式反应(Real-time PCR)检测B淋巴细胞瘤2(BCL-2)、人细胞凋亡调节因子(Bax)、人半胱氨酸蛋白酶3(Caspase-3)的表达情况。结果:4μmol/L组、8μmol/L组和16μmol/L组的细胞增殖抑制率、凋亡率和Caspase-3相对表达量水平均高于空白对照组,BCL-2/Bax相对表达量水平低于空白对照组,差异均有统计学意义(P<0.05)。随着斑蝥酸钠浓度的升高,人宫颈癌Hela细胞增殖抑制率、凋亡率和Caspase-3随之也升高,BCL-2/Bax相对表达量逐渐下降,差异均有统计学意义(P<0.05)。经斑蝥酸钠处理后可见细胞皱缩、变圆,细胞边缘出芽形成多花环结构的凋亡小体。结论:斑蝥酸钠具有抑制人宫颈癌Hela细胞增殖、促进其凋亡的作用,具有剂量依赖性,其对细胞凋亡的调控作用可能与调控BCL-2,Bax和Caspase-3表达有关。

Effect of aspirin on the proliferation and apoptosis of human adrenal cortical cancer cells

Objective To observe the effect of aspirin on the proliferation inhibition and apoptosis of human adrenal cortical cancer cells,and to explore preliminarily the related mechanisms.Methods The human adrenal cortical cancer cells were cultured in vitro.The experimental group was DEME/F-12 complete medium which contained different concentrations of aspirin(final concentration of 0.125,0.25,0.5,1 mg/ml).The control group was DEME/F-12 complete medium which had no aspirin but 1% anhydrous ethanol instead.After treatment by aspirin at different concentrations for different durations(24,48,72 hours),we detected the proliferation inhibition of SW-13 cells and H295R cells by methyl thiazolyl tetrazolium(MTT)method,observed the changes of cell morphology with the inverted microscope;Observed and counted apoptotic cells through Hoechst 33258 fluorescent staining,tested the cell apoptosis by flow cytometry,and detected the expression of Bcl-2 and Bax by western blotting.Results The date of MTT showed that after treated by different concentrations of aspirin,the growth inhibition ratio of SW-13 cells and H295R cells were higher than the control group(P<0.05),and at the same time,the inhibition ratio would increase when the drug concentration increased(P<0.05),with a dose-dependent tendency.When the drug concentration was constant,the inhibition ratio increased with the duration of the drug(P<0.05),which showed a time-dependent tendency.The number of apoptosis cells and the cell apoptosis rate of both SW-13 and H295R which were treated by different concentrations of aspirin for 48 hours were higher than those of the control group(P<0.01).According to the analysis of grayscale value of western blotting,aspirin can increase the expression of Bax(P<0.05).On the contrary,the expression of bcl-2 in the experimental group was lower than that in the control group(P<0.05).Conclusion Aspirin may inhibit the growth of human adrenal cortical cancer cell in vitro and induce its apoptosis,and the possible mechanism may be correlated with the up-regulation of the expression of Bax,and down regulation of Bcl-2 expression.

桑树类黄酮对三阴性乳腺癌细胞MDA-MB231的细胞毒性作用初探

桑树中富含的类黄酮具有多种药理活性。采用3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2H-四唑试剂(MTS)染色和实时荧光定量PCR(RT-qPCR)等方法,就桑树中类黄酮对乳腺癌细胞MDA-MB231的细胞毒性作用进行初步探索。实验结果显示,供试的4种桑树类黄酮中,戊烯基化类黄酮亚类对乳腺癌细胞的毒性作用强于非戊烯基化类黄酮。其中,戊烯基化类黄酮——桑根酮C整体呈现出更强的细胞毒性作用,当桑根酮C的浓度达到20μmol/L时能显著抑制MDA-MB231增殖。胸腺嘧啶脱氧核苷类似物(EDU)检测细胞增殖等实验结果显示,20μmol/L桑根酮C处理MDA-MB231后,细胞中DNA的复制明显减少,体外克隆能力显著减弱;RT-qPCR检测细胞周期阻滞和DNA损伤相关的周期蛋白依赖性激酶抑制剂1A基因(CDKN 1A)和生长阻滞与DNA损伤基因(GADD 45A)的转录水平显著上升,而DNA复制相关S期激酶相关蛋白2基因(SKP 2)和增殖细胞核抗原基因(PCNA)的转录水平则显著降低。此外,用桑根酮C处理MDA-MB231后,出现细胞形态变圆变亮和变小、细胞中线粒体膜电位丢失和ROS积累等现象,以及促凋亡基因phorbol-12-myristate-13-acetate诱导蛋白1基因(PMAIP 1)的转录水平显著上升,抑制凋亡基因Bcl 2的转录水平显著下降,提示桑根酮C可能诱导细胞发生凋亡。

2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮诱导Hep3B人肝癌细胞凋亡的机制

近年来,紫草萘醌类衍生物因其临床应用受到副作用的限制。为寻找副作用小疗效高的新型抗肿瘤药物,合成了2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮,并对其化学结构进行了鉴定。同时研究了2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮对人肝癌细胞活力、凋亡的影响及其潜在机制。研究结果表明,通过MTT检测其细胞活力,发现2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮显著降低了人肝癌细胞系的细胞活力。蛋白免疫印迹结果表明,该化合物可通过上调Cle-caspase3、Bad和Bax凋亡相关蛋白表达水平来诱导肝癌细胞Hep3B凋亡。为进一步检测2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮诱导Hep3B细胞发生凋亡的原因,使用荧光探针JC-1检测了2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮处理后Hep3B细胞内线粒体膜电位的变化。结果发现,与对照组相比,药物处理组线粒体膜电位显著下降,绿色荧光增强,红色荧光减弱,说明2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮能通过线粒体损伤来诱导Hep3B细胞凋亡。由于2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮显著诱导Hep3B细胞凋亡。因此,2-(4-甲氧基-苯巯基)-5,8-二甲氧基萘-1,4-二酮具有良好的抗肿瘤活性。

欣力康胶囊对血液性肿瘤细胞的增殖抑制作用研究

目的 探索欣力康胶囊在血液肿瘤上应用的可能性,以拓展欣力康胶囊的应用范围。方法 采用MTS比色法检测欣力康胶囊对8株人血液性肿瘤细胞的增殖抑制作用;采用流式细胞术检测欣力康胶囊对急性髓系白血病MV-4-11细胞周期阻滞和凋亡促进的作用;采用蛋白免疫印迹法检测欣力康胶囊对MV-4-11细胞内pro-Caspase 3、cleaved-Caspase 3以及NF-κB p65蛋白水平的影响;依托MV-4-11皮下移植瘤裸小鼠模型,考察欣力康胶囊对荷瘤小鼠肿瘤生长的影响。结果 欣力康胶囊对MV-4-11、RS4;11、MM.1S、RPMI-8226、Mino、Jeko-1、OCI-LY10和TMD8细胞增殖抑制的IC_(50)值分别为(54.05±6.33)、(125.33±2.46)、(655.87±7.95)、(153.33±8.50)、(188.70±9.11)、(180.87±7.31)、(186.10±6.55)、(253.53±8.88)μg·mL^(-1);欣力康胶囊可促进MV-4-11细胞凋亡,且具有浓度依赖性,当质量浓度达到500 μg·mL^(-1)时凋亡率为(99.41±0.36)%(P<0.001),而对细胞周期影响较弱;欣力康胶囊可促进MV-4-11细胞中凋亡相关蛋白pro-Caspase 3的剪切,下调NF-κB p65,且呈一定的浓度依赖性;临床用药浓度(1000 mg·kg^(-1))的欣力康胶囊能够显著抑制荷MV-4-11皮下移植瘤小鼠的肿瘤生长,其抑瘤率为57.28%。结论 欣力康胶囊对8种血液性肿瘤细胞株的增殖均有一定的抑制作用,其中对人急性髓性白血病MV-4-11细胞的增殖抑制作用最优,其机制主要是抑制NF-κB通路以及促进细胞凋亡,且在体内药效上,临床用药浓度的欣力康胶囊可显著抑制荷MV-4-11皮下移植瘤小鼠肿瘤的生长。以上结果提示欣力康胶囊可用于血液性肿瘤的辅助治疗。

枸杞外泌体对非小细胞肺癌A549细胞的增殖抑制和凋亡的调控作用

为阐明枸杞外泌体对非小细胞肺癌细胞A549的增殖抑制和凋亡的调控作用,通过超速离心法提取枸杞外泌体,通过透射电镜检测外泌体的形态,粒径分析检测外泌体的直径分布。用提取到的枸杞外泌体作用于A549细胞,通过CCK-8法检测枸杞外泌体对A549细胞的增殖影响,EdU检测枸杞外泌体对A549细胞的活性影响,细胞划痕、Transwell检测细胞的迁移效率,细胞克隆法检测细胞的克隆能力。流式细胞术检测细胞的凋亡率,用Western Blot和qRT-PCR法检测细胞凋亡通路的蛋白和基因表达。结果表明,提取的枸杞外泌体在电镜下呈茶托状,大小在40~200 nm,符合外泌体形状特征。与对照组相比,枸杞外泌体组能够浓度依赖性地抑制A549细胞增殖(P<0.01)。不同浓度枸杞外泌体处理组的细胞迁移能力和细胞克隆能力也呈浓度依赖性下降(P<0.01),而细胞凋亡率呈浓度依赖性增多(P<0.01)。枸杞外泌体能够浓度依赖性地上调Cl-Caspase3、p53、Bax等促凋亡因子,下调Bcl-2等抑凋亡因子的蛋白和基因(P<0.01)。结果表明,枸杞外泌体可以抑制A549细胞增殖,促进A549细胞凋亡并且枸杞外泌体可通过线粒体通路诱导细胞凋亡。

基于自噬抑制剂3-MA介导的大叶蛇葡萄总黄酮提取物对人乳腺癌细胞增殖、凋亡的影响

目的探讨抑制自噬时,大叶蛇葡萄总黄酮提取物(total flavonoid extract,TFE)对人乳腺癌细胞增殖、凋亡的影响及其作用机制。方法针对人宫颈癌细胞Hela、人肺癌细胞A549、人肝癌细胞SMMC-7721、人乳腺癌细胞MCF-7、MDA-MB-231及人正常肝细胞L-02,采用MTT法优选敏感细胞株;通过TFE与自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)联合运用,采用MTT法检测其对敏感细胞增殖的抑制作用;透射电子显微镜及Hoechst 33258单染法观察细胞的形态学变化;Annexin V-FITC/PI双染法检测细胞凋亡率的变化;Western blot检测凋亡相关蛋白和通路蛋白(死亡受体途径、线粒体途径、内质网应激途径)的表达水平;免疫荧光法检测线粒体途径关键蛋白Cyt-c表达,并选择自噬激动剂雷帕霉素(rapamycin,RA)进行反向验证。结果TFE可浓度依赖性抑制人乳腺癌细胞增殖,MCF-7细胞为敏感细胞株,与TFE组相比,TFE+3-MA组在24、48、72 h对MCF-7细胞的抑制率均明显增加(P<0.01),细胞数量减少、间隙增大,凋亡小体增多,凋亡率升高(P<0.01),Bax/Bcl-2(P<0.01)、cleaved caspase-3(P<0.01)、Cyt-c(P<0.05)、FADD、cleaved caspase-12的表达量均升高,核内凋亡蛋白Cyt-c表达增加;TFE+RA组荧光减弱,逆转了TFE诱导的线粒体途径凋亡。结论TFE能明显抑制人乳腺癌细胞的增殖,抑制自噬时可能通过线粒体途径促进MCF-7细胞凋亡,激活自噬可逆转细胞凋亡。

大豆黄素通过巨噬细胞移动抑制因子抑制肝癌细胞增殖

[目的]探讨大豆黄素(Glycitein)通过巨噬细胞移动抑制因子(migration inhibitory factor,MIF)抑制肝癌细胞增殖的机制。[方方法]采用生物信息学分析大豆黄素作用靶点,并分析该靶点在肝癌组织中表达及其与临床不良表型之间的关系。将肝癌细胞(HepG2和HepB3)分成正常组、Glycitein组(Glycitein处理)和Glycitein+MIF组(转染MIF并用Glycitein处理)。采用不同浓度Glycitein干预,以细胞增殖实验检测细胞活力;细胞克隆实验检测Glycitein对细胞生长能力的影响;细胞周期实验检测Glycitein对细胞周期的影响;细胞凋亡实验检测Glycitein对细胞凋亡的影响;裸鼠移植瘤实验检测Glycitein体内抑制细胞增殖;免疫组化检测增殖细胞抗原Ki67;免疫印迹法检测细胞MIF以及凋亡相关蛋白表达。[结果]MIF是Glycitein作用靶点,在肝癌组织中高表达,且与肝癌临床不良表型有关。Glycitein作用后,能够抑制肝癌细胞增殖和细胞周期进程,并诱导细胞凋亡。裸鼠移植瘤实验显示,Glycitein作用裸鼠后,明显抑制移植瘤体积和质量增长。Glycitein组细胞Ki67相对表达量明显低于正常组(P<0.05)。Glycitein能够呈浓度和时间依赖性地抑制肝癌细胞MIF蛋白表达。与Glycitein+MIF-NC组比较,Glycitein+MIF组细胞增殖、克隆增高,而G0/G1期细胞比例减低,凋亡细胞数量减少(P<0.05)。与Glycitein+MIF-NC组比较,Glycitein+MIF组细胞B细胞淋巴瘤/白血病-2(B cell lymphoma/leukemia-2,Bcl-2)、细胞周期蛋白依赖性激酶2(cyclin-dependent kinases 2,CDK2)、细胞周期蛋白B1(cyclin B1,CCNB1)和Ki67蛋白表达增高(P<0.05),Bcl-2相关X蛋白(Bcl-2 related X,Bax)蛋白表达减低(P<0.05)。[结论]Glycitein通过靶向调控MIF蛋白表达,抑制肝癌细胞增殖,阻滞细胞周期,促进细胞凋亡。

5种新疆香阿魏对结肠癌细胞增殖抑制、促凋亡及逆转耐药作用的研究

通过MTT法和流式细胞术检测5种新疆香阿魏(准噶尔阿魏、多伞阿魏、山地阿魏、全裂叶阿魏、荒地阿魏)乙醇提取物对小鼠结肠癌CT-26.WT细胞的增殖抑制及促凋亡作用,并通过MTT法检测5种新疆香阿魏乙醇提取物对人结肠癌耐长春新碱HCT-8/VCR细胞的安全给药浓度及逆转耐药作用。结果发现,5种新疆香阿魏乙醇提取物均对小鼠结肠癌CT-26.WT细胞有增殖抑制作用;全裂叶阿魏、准噶尔阿魏乙醇提取物的抑制作用较好,IC_(50)值分别为20.93 mg·L^(-1)、22.19 mg·L^(-1)。全裂叶阿魏乙醇提取物对小鼠结肠癌CT-26.WT细胞有较明显的促凋亡作用,凋亡率达到90.26%。5种新疆香阿魏乙醇提取物均可抑制HCT-8/VCR细胞的增殖,其中逆转耐药作用最佳的是准噶尔阿魏乙醇提取物。表明,全裂叶阿魏、准噶尔阿魏对CT-26.WT细胞的增殖抑制、促凋亡作用较明显,并且准噶尔阿魏对HCT-8/VCR细胞具有较好的逆转耐药作用,准噶尔阿魏、全裂叶阿魏均具有一定的抗肿瘤效果,值得更进一步的深入研究。