单纯疱疹病毒胸苷激酶基因治疗结肠癌的研究

目的：观察单纯疱疹病毒胸苷激酶(HSV-tk),丙氧鸟苷(GCv)自杀基因治疗系统在体内外对结肠癌的杀伤效应。方法：构建携带HSV-tk基因的逆转录病毒载体,将该重组逆转录病毒感染结肠癌细胞株SW480,筛选稳定表达tk的细胞克隆SW480／tk,测定SW480／tk细胞在体外对GCV的敏感性;SW480／tk细胞在裸鼠皮下成瘤,用GCV治疗并观察疗效。结果：HSV-tk基因整合入SW480细胞并在SW480／tk细胞中稳定表达;GCV在体外对SW480／tk细胞有明显的杀伤作用,其作用呈现剂量和时间依赖性特点和旁杀伤效应,对未转染细胞则无明显毒性。裸鼠ex vivo实验得到相应结果,治疗组肿瘤受到明显抑制。结论 在in vitra和ex vivo水平,表达HSV-tk基因的肿瘤细胞均可被GCV有效杀伤,逆转录病毒介导HSV-tk/GCV自杀基因治疗系统有可能成为结肠癌的有效治疗方法。     

奥沙利铂对人结肠癌细胞凋亡及FADD的影响

以不同浓度（0、25、50和100μg/ml）的奥沙利铂干预体外培养的人结肠癌细胞株（SW480）,分别于12、24和48 h后,应用四甲基偶氮唑盐比色法检测细胞的增殖活性,流式细胞仪检测细胞周期和细胞凋亡率,半定量RT-PCR检测Fas相关死亡结构域蛋白（FADD）mRNA水平,Western blotting检测FADD蛋白水平。结果奥沙利铂对SW480生长增殖具有明显抑制作用,其效应呈浓度和时间依赖性,SW480细胞呈G2/M期阻滞;SW480细胞的凋亡率呈浓度依赖性的增加（P〈0.05）。不同浓度的奥沙利铂处理SW480细胞株24 h后,FADD mRNA和蛋白水平呈浓度依赖性增加。提示奥沙利铂能诱导SW480细胞凋亡,其作用机制可能与其激活凋亡死亡受体途径使FADD表达上调有关。     

CpG ODN对食管癌细胞RNA转染的脐血树突状细胞诱导CTL抗肿瘤功能的影响

目的探讨食管癌细胞RNA转染脐血树突状细胞（DCs）对细胞毒性T淋巴细胞（CTL）特异性抗肿瘤作用的影响,以及CpG寡脱氧核糖核苷酸（CpG ODN）的免疫佐剂作用。方法分离脐血单个核细胞,经SCF、GM-CSF和IL-4诱导和食管癌RNA转染形成成熟DCs,加入CpG ODN,检测DCs表面标志及DCs诱导的T细胞增殖、CTL杀伤活性。结果食管癌RNA转染后DCs高表达抗原提呈分子、协同刺激分子和黏附分子,对T细胞的促增殖作用和CTL杀伤活性显著增强（P〈0.01）,CpG ODN能显著促进DCs功能。结论CpG ODN具有增强食管癌细胞RNA转染的DCs对T细胞的促增殖作用和促CTL杀伤活性的作用。     

芥菜籽提取物对人结肠癌细胞系SW480凋亡的影响

目的观察芥菜籽提取物（MSE）对人结肠癌细胞系SW480细胞凋亡的影响并初步探讨其可能的机制。方法采用体外细胞培养技术,用CCK-8法观察MSE对人大肠癌SW480细胞生长的影响;Hoechest3325荧光染色显微镜下观察凋亡细胞形态学改变,流式细胞仪检测细胞凋亡;Caspase-3活性检测试剂盒分析其凋亡机制。结果在浓度0．2—1．0mg／ml作用24—72h范围内,MSE对SW480细胞增殖具有明显抑制作用（P〈0．01）。24h内其凋亡率随着药物浓度的增加而增加,且逐渐从早期凋亡走向晚期凋亡,呈浓度依赖性;24h内Caspase-3活性较对照组显著提高（P〈0．05）。结论MSE能有效抑制体外培养的人结肠癌细胞SW480的生长,这种抑制作用与MSE促进肿瘤细胞凋亡、激活凋亡蛋白酶Caspase-3的活性有关。     

慢性乙型重型肝炎患者外周血单个核细胞B7/CD28分子的表达及意义

为了解慢重型肝炎肝损伤中B7／CD28共刺激途径的作用，我们应用流式细胞技术（FCM），检测慢性乙型重型肝炎（CSHB）及慢性乙型肝炎（CHB）患者外周血单个核细胞（PBMC）中淋巴细胞和单核细胞上表达B7／CD28共刺激分子的情况，并探讨其临床意义。     

DC与CIK或LAK联合对结肠癌细胞株SW480杀伤作用的研究

[目的]研究树突状细胞(DC)联合细胞因子诱导或未诱导的杀伤细胞(CIK)或淋巴因子激活的杀伤细胞(LAK)对结肠癌细胞株SW480的杀伤活性.提供DC联合CIK或LAK治疗结肠癌的实验依据.[方法]取人外周血分离出单个核细胞(PBMNC),诱导生成DC、CIK、LAK细胞;流式细胞仪检测DC经SW480肿瘤抗原冲击后的表型变化;以CIK+DC细胞、CIK细胞、LAK+DC细胞及LAK细胞作为效应细胞,SW480为靶细胞,以15∶1、30∶1、45∶1为效靶比,LDH释放法测定细胞杀伤试验活性;ELISA检测杀伤试验中干扰素γ(IFN-γ)、白细胞介素2(IL-2)、IL-12、IL-17的分泌水平.[结果]流式细胞仪检测DC经SW480肿瘤抗原冲击后,其表面分子HLA-DR、CD40、CD80和CD86表达分别平均为90.23％、73.68％、85.96％、57.55％,与未经肿瘤抗原冲击DC比较,DC成熟的表面标志分子表达明显增加(P＜0.01).相同效靶比下,CIK+DC细胞组对SW480的杀伤作用最强,明显高于其他细胞组(P＜0.01);CIK+ DC细胞组在效靶比为45∶1时,杀伤活性最强(P＜0.01);单独CIK细胞组的杀伤活性明显高于LAK+DC细胞组(P＜0.01);LAK+ DC细胞组的杀伤活性明显高于单独LAK细胞组(P＜0.01).效靶比为45∶1时,各杀伤试验细胞组上清液中IFN-γ、IL-2、IL-12、IL-17的分泌量,CIK+DC细胞组的IFN-γ、IL-12的分泌量显著高于其他细胞组(P＜0.05);LAK+DC、单独LAK细胞组IL-2的分泌量明显高于CIK+DC、单独CIK细胞组(P＜0.05);单独CIK细胞组IFN-γ的分泌量明显高于LAK+DC、单独LAK细胞组(P＜0.05).[结论]CIK+DC细胞组对SW480的杀伤活性明显强于单独CIK、LAK+ DC组、单独LAK细胞组.其机制可能是,SW480抗原致敏的DC分泌IFN-γ、IL-12等刺激、诱导CIK细胞的活化和增殖,明显增强CIK细胞杀伤SW480的活性.     

胃泌素在结肠癌患者中的表达及其受体拮抗剂对人结肠癌细胞株的抑制作用及其对P38信号转导通路的影响

背景结肠癌(colon cancer, CC)是我国常见消化系统恶性肿瘤,早期缺乏特异性症状诊断率较低,导致患者丧失根治性机会,病死率较高,极大危害患者生命健康.胃泌素主要是由胃肠道G细胞分泌一种激素,与胃泌素受体结合后可刺激胃酸分泌,促进胃肠道黏膜生长.丝裂原活化蛋白激酶是一组能被胃泌素等激素激活的丝氨酸-苏氨酸蛋白激酶,负责细胞表面与细胞核内部间信号传递.目的分析胃泌素在CC患者中的表达,并探讨其受体拮抗剂对人CC细胞株的抑制作用及对P38信号转导通路的影响.方法将2016-01/2018-10我院病理科30例CC组织标本根据世界卫生组织恶性肿瘤分化程度标准分为低分化标本、中分化标本、高分化标本,采用免疫组化技术检测观察胃泌素在CC组织中表达情况,体外培养人CC细胞株SW480,将细胞分为对照组(不进行任何药物处理)、胃泌素组(分别加入6.25-200.00 mg/L胃泌素进行处理)、丙谷胺组(分别加入8.00-256.00 mg/L丙谷胺进行处理)、胃泌素联合丙谷胺组(加入12.5 mg/L胃泌素与8.00-256.00 mg/L丙谷胺进行处理),统计SW480中胃泌素受体/胆囊收缩素-B受体表达情况,比较各组CC细胞株SW480活力、细胞增殖指数、P38信号转导通路表达情况P38蛋白、磷酸化-P38蛋白、B淋巴细胞瘤-2(B lymphocyte tumor-2, SBcl-2)、细胞凋亡促进基因(BAX).结果 CC组织分化程度越高,胃泌素表达阳性率越高;胃泌素组6.25-200.00 mg/L范围内SW480 OD值均高于对照组(P0.05);胃泌素组12.50 mg/L时SW480 OD值最高(P0.05);胃泌素组25.00-200.00 mg/L范围内SW480OD值比较差异无统计学意义(P0.05);丙谷胺组8.00-256.00 mg/L范围内SW480 OD值比较差异无统计学意义(P0.05);胃泌素组联合丙谷胺组在12.50mg/L胃泌素联合16.00mg/L丙谷胺时,SW480O D值最低,低于对照组(P 0.05),之后随着丙谷胺浓度增加,SW480OD值比较差异无统计学意义(P0.05);胃泌素组(12.50mg/L)细胞增殖指数高于丙谷胺组(16.00 mg/L)、胃泌素组联合丙谷胺组(12.5mg/L+16.00 mg/L)(P 0.05);胃泌素组(12.50 mg/L)P38蛋白、磷酸化-P38蛋白、BAX蛋白低于对照组、丙谷胺组(16.00 mg/L)、胃泌素组联合丙谷胺组(12.5mg/L+16.00mg/L),Bcl-2蛋白表达高于对照组、丙谷胺组(16.00 mg/L)、胃泌素组联合丙谷胺组(12.5 mg/L+16.00 mg/L)(P 0.05).结论胃泌素可抑制人CC细胞株SW480的凋亡,且在CC组织中的表达与肿瘤分化程度有关,分化程度越高,其表达量越高,胃泌素受体拮抗剂在一定浓度范围内可拮抗胃泌素促增殖效应,其机制与上调P38、磷酸化-P38、BAX表达及下调Bcl-2表达有关.     

Fas通路的激活促进结肠癌细胞迁移侵袭

目的探讨Fas通路激活对结肠癌细胞SW480和DLD1产生的非凋亡效应。方法流式细胞学检测结肠癌SW480及DLD1细胞系的Fas、FasL、抗凋亡蛋白c-FLIP、Bcl-2、Bcl-xl、XIAP等在细胞膜上的表达水平;对结肠癌SW480及DLD1细胞系予以梯度浓度（0ng/ml、12.5ng/ml、25ng/ml、50ng/ml）的FasL刺激,计数并比较各组细胞的增殖速率变化;对结肠癌SW480及DLD1细胞系予以低剂量FasL处理,分别对实验组和对照组进行细胞增殖速率及迁移侵袭能力测试,探讨低剂量FasL刺激对结肠癌细胞活力的影响;稳定敲除SW480和DLD1细胞的Fas基因表达,重复上述实验,以明确低剂量FasL刺激对结肠癌细胞活力的影响是否依赖于Fas通路的激活。结果 SW480及DLD1细胞系中均可检测到中等程度的Fas表达及抗凋亡蛋白FLIP、Bcl-2的表达,未检测到FasL表达,在SW480中可测到Bcl-xl表达;低剂量FasL不影响结肠癌SW480和DLD1细胞的增殖速率,但可促进两种细胞的迁移侵袭能力;稳定敲除Fas基因后,低剂量FasL对结肠癌细胞迁移侵袭能力的促进作用受到抑制。结论低剂量FasL可激活Fas通路的非凋亡途径从而增强结肠癌SW480和DLD1细胞的迁移侵袭能力。     

姜黄素对人结肠癌细胞SW480作用的体外实验研究

目的 研究姜黄素促进人结肠癌细胞系SW480凋亡的作用.方法 人结肠癌细胞系SW480与不同浓度的姜黄素共孵育48 h,MTT法检测细胞存活率,并用相差显微镜观察细胞形态学改变.用流式细胞术(检测细胞凋亡相关指标Annexin V/PI)和Ho-echst3258染色检测细胞凋亡.结果 姜黄素能够不同程度地抑制人结肠癌细胞系SW480的生长.姜黄素对SW480细胞的IC50(半数抑制浓度)为65 mg/mL.Hoechst3258荧光染色与流式细胞术分析SW480细胞的结果一致.结论 姜黄素可促进SW480细胞凋亡,为姜黄素治疗结肠癌提供依据.     

NF-κB decoy寡核苷酸对结肠癌SW480细胞TLR4表达的影响

目的 探讨核转录子κB(NF-κB)decoy寡核苷酸(ODNs)对结肠癌SW480细胞Toll样受体4(TLR4)表达的影响. 方法 体外培养SW480细胞,LPS(10μL)刺激3 h后,用脂质体Lipofectin2000介导NF-κB decoy ODNs转染6 h,RT-PCR法检测细胞的TLR4 mRNA.设对照组、LPS组、SODN组、Lipofectin2000组. 结果 转染NF-κB decoy ODN的SW480细胞的TLR4 mRNA表达明显高于对照组(P<0.01),但明显低于其他组(P均<0.01). 结论 NF-κB decoy ODNs明显抑制SW480细胞TLR4的表达.     

NK cells stimulated with IL-15 or CpG ODN enhance rituximab-dependent cellular cytotoxicity against B-cell lymphoma

OBJECTIVE: Antibody-dependent cellular cytotoxicity (ADCC) is an important mechanism in the clinical activity of rituximab for treatment of B-cell malignancies. Natural killer (NK) cells, through the activating receptor FcgammaRIIIa (CD16), play a major role in rituximab-mediated ADCC. We have studied the in vitro effect of NK stimulators, such as interleukin-15 (IL-15) and CpG oligodeoxynucleotides A-Class (CpG ODN A), in the enhancement of rituximab-mediated ADCC against B-cell lymphoma. METHODS: Peripheral blood mononuclear cells (PBMC), purified NK cells, or NK-depleted PBMC from healthy donors, were activated with IL-15 or CpG ODN A, and cocultured with B-lymphoma cells in the presence of rituximab to evaluate the enhancement of the cytotoxicity. RESULTS: The rituximab-mediated ADCC of IL-15-activated PBMC was twofold compared to unstimulated PBMC (73% +/- 7% vs 37% +/- 5% respectively, p < 0.001). Similarly, rituximab-mediated ADCC was enhanced when PBMC were activated with CpG ODN A as compared to CpG ODN control (61% +/- 11% vs 36% +/- 8%, respectively, p = 0.02). Nevertheless, the ADCC of purified NK cells was increased only with IL-15. NK-depleted PBMC activated with either IL-15 or CpG ODN A showed no ADCC, suggesting that NK are the major effector cells. Furthermore, IL-15 or CpG ODN A-activated PBMC, but not activated purified NK cells, secreted large amounts of interferon-gamma in the presence of rituximab-coated lymphoma cells. CONCLUSIONS: IL-15 and CpG ODN A enhance rituximab-mediated ADCC against B-cell lymphoma. Under these conditions, NK cells seem to be the main effector cells mediating ADCC. These findings suggest that these agents could be used as adjuvants in combination with rituximab for patients with B-cell lymphoma.     

藤梨根提取物通过调控miR-192-5p/ARPP19轴影响结直肠癌细胞的增殖和凋亡

背景 藤梨根提取物(rattan root extract,RRE)可抑制胃癌、肺癌等肿瘤细胞增殖,具有一定抗肿瘤作用,但其是否影响结直肠癌细胞的恶性表型还未知.miR-192-5p在结直肠癌组织中表达降低,且其低表达与肿瘤大小等临床病理特征相关,可作为结直肠癌诊治的潜在生物学标志物.StarBase生物信息学软件预测...     

Synergistic antitumor effect of TRAIL and doxorubicin on colon cancer cell line SW480

AIM: TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) has been reported to specifically induce apoptosis of cancer cells although only a small percentage of cell lines were sensitive to it. Cell lines not responding to TRAIL in vitro were said to be more prone to apoptosis when TRAIL was combined with another anticancer agent.Generally, factors affecting drug-sensitivity involve many apoptosis-related proteins, including p53. The expression of wild-type p53 gene was proposed as an important premise for tumor cells responding to chemotherapy. The present study was to investigate the cell killing action of TRAIL on colon cancer cell line SW480, its synergistic effect with doxorubicin, and the possible mechanisms.METHODS: SW480 cells were cultured in the regular condition and incubated with different levels of agents.Morphologic changes in these cells after treatment were observed under phase-contrast microscope and cytotoxicity by TRAIL alone and in combination with doxorubicin was quantified by a 1-day microculture tetrazolium dye (MTT) assay. In addition, flow cytometry assay (FCM) and transmission electron microscopy were used to detect apoptosis among these cells. Variation of p53 protein level among different groups according to concentrations of agents was measured by Western blot assay.RESULTS: (1) SW480 cells were not sensitive to TRAIL,with IC50&gt;l mg&#183;L^1 and dose-independent cytotoxicity. (2)SW480 cells were sensitive to doxorubicin at a certain degree,with dose-dependent cytotoxicity and IC50=65.25&#177;3.48μmol&#183;L^-1. (3) TRAIL could synergize with doxorubicin to kill SW480 cells effectively, which was represented by the boosted killing effect of doxorubicin on theses cells. IC50 of doxorubicin against SW480 cells sharply reduced when it was combined with TRAIL. (4) Subtoxic TRAIL (100 μg&#183;L^-1),combined with subtoxic doxorubicin (0.86 μmol&#183;L^-1), could kill SW480 cells sufficiently. Cytotoxicity by MTT assay arrived at 80.12&#177;2.67 %, which was significantly higher than that by TRAIL or doxorubicin alone, with P=0.006 and 0.003 respectively. This killing effect was partly due to apoptosis. It was proved by large amounts of apoptotic cells under phase-contrast microscopy, cell apoptosis rate of 76.82&#177;1.93 % by FCM assay and typical apoptotic morphology observed through transmission electron microscopy. Increase of apoptosis after combined treatment had no relation with protein level of p53 (p&gt;0.05).CONCLUSION: SW480 cells are not sensitive to TRAIL, but TRAIL can synergize with lower concentra~on of doxorubidn to induce apoptosis effectively. The status of p53 protein is not involved in the mechanism of synergistic apoptosis. It suggests the potential therapeutic applicability of the combination of TRAIL with doxorubidn against colon cancers.     

Synergistic antitumor effect of TRATL and doxorubicin on colon cancer cell line SW480

AIM: TRAIL (tumor necrosis factor-related apoptosisinducing ligand) has been reported to specifically induce apoptosis of cancer cells although only a small percentage of cell lines were sensitive to it. Cell lines not responding to TRAIL in vitro were said to be more prone to apoptosis when TRAIL was combined with another anticancer agent.Generally, factors affecting drug-sensitivity involve many apoptosis-related proteins, including p53. The expression of wild-type p53 gene was proposed as an important premise for tumor cells responding to chemotherapy. The present study was to investigate the cell killing action of TRAIL on colon cancer cell line SW480, its synergistic effect with doxorubicin, and the possible mechanisms. METHODS: SW480 cells were cultured in the regular condition and incubated with different levels of agents.Morphologic changes in these cells after treatment were observed under phase-contrast microscope and cytotoxicity by TRAIL alone and in combination with doxorubicin was quantified by a 1-day microculture tetrazolium dye (MTT)assay. In addition, flow cytometry assay (FCM) and transmission electron microscopy were used to detectapoptosis among these cells. Variation of p53 protein level among different groups according to concentrations of agents was measured by Western blot assay.RESULTS: (1) SW480 cells were not sensitive to TRAIL,with IC50＞1 mg@L1 and dose-independent cytotoxicity. (2)SW480 cells were sensitive to doxorubicin at a certain degree,with dose-dependent cytotoxicity and IC50=65.25+3.48μmol@L-1. (3) TRAIL could synergize with doxorubicin to kill SW480 cells effectively, which was represented by the boosted killing effect of doxorubicin on theses cells. IC50 of doxorubicin against SW480 cells sharply reduced when it was combined with TRAIL. (4) Subtoxic TRAIL (100 μg.L-1),combined with subtoxic doxorubicin (0.86 μmol@L1), could kill SW480 cells sufficiently. Cytotoxicity by MTT assay arrived at 80.12+2.67%, which was significantly higher than that by TRAIL or doxorubicin alone, with P=0.006 and 0.003 respectively. This killing effect was partly due to apoptosis. It was proved by large amounts of apoptotic cells under phase-contrast microscopy, cell apoptosis rate of 76.82±1.93 % by FCM assay and typical apoptotic morphology observed through transmission electron microscopy. Increase of apoptosis after combined treatment had no relation with protein level of p53 (P＞0.05). CONCLUSION: SW480 cells are not sensitive to TRAIL, but TRAIL can synergize with lower concentration of doxorubicin to induce apoptosis effectively. The status of p53 protein i snot involved in the mechanism of synergistic apoptosis. It suggests the potential therapeutic applicability of the combination of TRAIL with doxorubicin against colon cancers.     

Mechanisms underlying aspirin-mediated growth inhibition and apoptosis induction of cyclooxygenase-2 negative colon cancer cell line SW480

AIM： To investigate the effects of aspirin （acetylsalicylic acid） on proliferation and apoptosis of colorectal cancer cell line SW480 and its mechanism. METHODS： Cyclooxygenase （COX）-2 negative colorectal cancer cell line SW480 was treated with aspirin at concentrations of 2.5 mmol/L, 5.0 mmol/L, 10.0 mmol/L for different periods in v/tro. Anti-proliferation effect of aspirin on SW480 was detected by 3-（4,5-dimeth- ylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay. Cell cycle and apoptosis were observed by flow cytometry （FCM）. Transmission electron microscope （TEM） was used for morphological study. Apoptosis-associated genes were detected by immunohistochemical staining and Western blotting. RESULTS： Aspirin inhibited SW480 proliferation and induced apoptosis in a dose- and time-dependent manner. Treatment with different concentrations of aspirin significantly increased the proportions of cells at the G0/G1 phase and decreased the proportions of cells at the S- and G2/M phases in a concentration-dependent manner. Aspirin not only induced apoptosis but also caused cell necrosis at a high concentration as well. After treatment with aspirin, SW480 cells displayed typically morphological features of apoptosis and necrosis under TEM, and increased the Bcl-2 expression in cells, but the expression of Bax was down regulated. CONCLUSION： Aspirin inhibits proliferation and induces apoptosis of SW480 cells. Its anti-tumor mechanism may arrest cell cycle and shift Bax/Bcl-2 balance in cells.     

Effect of antisense oligodeoxynucleotide of telomerase RNA on telomerase activity and cell apoptosis in human colon cancer

AIM:To explore the effect of antisense oligocleoxynucletide (As-ODN) of telomerase RNA on telomerase activity and cell apoptosis in human colon cancer.RESULTS:The telomerase activity in SW480 cells transfected with 1.0μmol/L of As-ODN for 2-5days, was significantly decreased in a time-dependent manner, and the cells underwent apoptosis.The missense ODN (Ms-ODN) and the control group transfected with SW480 cells did not show these changes.CONCLUSION:As-ODN can specifically inhibit the telomerase activity of SW480 cells and induce apoptosis.     

EGCG对LoVo和SW480结肠癌细胞株的作用及对Notch1与Notch2基因表达的影响

目的:探讨绿茶提取物表没食子儿茶素没食子酸酯(EGCG)对结肠癌细胞株LoVo细胞和SW480细胞增殖的抑制作用,研究其对Notch1与Notch2的基因表达的影响,方法:体外培养LoVo细胞和SW480细胞,采用不同浓度的EGCG(10、20、35 mg/L)对其进行干预,MTT法检测EGCG对LoVo细胞和SW48...     

Inhibitory effect of caffeic acid phenethyl ester on the growth of SW480 colorectal tumor cells involves β-catenin associated signaling pathway down-regulation

AIM: To study the anti-tumor effect of caffeic acid phenethyl ester (CAPE) and the influence of CAPE on β-catenin associated signaling pathway in SW480colorectal cancer (CRC) cells.METHODS: SW480 cells were treated with CAPE at serial concentrations. The proliferative status of cells was measured by methabenzthiazuron (MTT) assay. Cell cycle and cell apoptosis were analyzed using flow cytometry (FCM). Western blotting assay was used to evaluate the protein level of β-catenin, c-myc and cyclinD1.β-catenin localization was determined by indirect immunofluorescence.RESULTS: CAPE displayed a strong inhibitory effect in a significant dose- and time-dependent manner on SW480cell growth. FCM analysis showed that the ratio of G0/G1 phase cells increased, S phase ratio decreased and apoptosis rate increased after SW480 cells were exposed to CAPE for 24 h. Pretreatment of SW480 cells with CAPE significantly suppressed β-catenin, c-myc and cyclinD1protein expression. CAPE treatment was associated with decreased accumulation of β-catenin protein in nucleus and cytoplasm, and concurrently increased its accumulation on the surface of cell membrane.CONCLUSION: CAPE can inhibit SW480 cell proliferation by inducing cell cycle arrest and apoptosis. Decreased β-catenin and the associated signaling pathway target gene expression may mediate the anti-tumor effects of CAPE.     

Inhibitory effect of caffeic acid phenethyl ester on the growth of SW480 colorectal tumor cells involves β-catenin associated signaling pathway down-regulation

AIM: To study the anti-tumor effect of caffeic acid phenethyl ester (CAPE) and the influence of CAPE on beta-catenin associated signaling pathway in SW480 colorectal cancer (CRC) cells. METHODS: SW480 cells were treated with CAPE at serial concentrations. The proliferative status of cells was measured by methabenzthiazuron (MTT) assay. Cell cycle and cell apoptosis were analyzed using flow cytometry (FCM). Western blotting assay was used to evaluate the protein level of beta-catenin, c-myc and cyclinD1. Beta-catenin localization was determined by indirect immunofluorescence. RESULTS: CAPE displayed a strong inhibitory effect in a significant dose- and time-dependent manner on SW480 cell growth. FCM analysis showed that the ratio of G0/G1 phase cells increased, S phase ratio decreased and apoptosis rate increased after SW480 cells were exposed to CAPE for 24 h. Pretreatment of SW480 cells with CAPE significantly suppressed beta-catenin, c-myc and cyclinD1 protein expression. CAPE treatment was associated with decreased accumulation of beta-catenin protein in nucleus and cytoplasm, and concurrently increased its accumulation on the surface of cell membrane. CONCLUSION: CAPE can inhibit SW480 cell proliferation by inducing cell cycle arrest and apoptosis. Decreased beta-catenin and the associated signaling pathway target gene expression may mediate the anti-tumor effects of CAPE.