丙戊酸对人脑胶质母细胞瘤细胞株抑制增殖及诱导凋亡作用研究

目的研究丙戊酸(2-propylpentanoic acid,VPA)体外对人脑胶质母细胞瘤细胞株增殖抑制、诱导细胞周期阻滞及促进凋亡的作用,为临床治疗提供理论依据。方法 MTT比色法检测VPA对胶质母细胞瘤细胞株的杀伤作用;流式细胞术检测其对细胞周期及凋亡的影响作用;Western blot法检测乙酰化组蛋白H3(acetyl-Histone H3)、乙酰化组蛋白H4(acetyl-histone H4)表达量变化情况。结果 VPA对胶质瘤母细胞瘤细胞株SF295、U87具有抑制增殖作用,诱导细胞周期阻滞于G2/M期,乙酰化组蛋白H3、H4表达量呈时间依赖性增加。大剂量可以诱导出现凋亡。结论 VPA能够在体外抑制胶质瘤细胞生长,诱导细胞周期阻滞及促进细胞凋亡,其机制可能与促进组蛋白乙酰化有关。     

Naringin suppresses the development of glioblastoma by inhibiting FAK activity

As the most common and lethal primary malignant brain cancer, glioblastoma is hard to timely diagnose and sensitive therapeutic monitoring. It is essential to develop new and effective drugs for glioblastoma multiform. Naringin belongs to citrus flavonoids and was found to display strong anti-inflammatory, antioxidant and antitumor activities. In this report, we found that naringin can specifically inhibit the kinase activity of FAK and suppress the FAK^p-Try397and its downstream pathway in glioblastoma cells. Our study showed out that naringin can inhibit cell proliferation by inhibiting FAK/cyclin D1 pathway, promote cell apoptosis through influencing FAK/bads pathway, at the same time, it can also inhibit cell invasion and metastasis by inhibiting the FAK/mmps pathway. All these showed that naringin exerts the anti-tumor effects in U87?MG by inhibiting the kinase activity of FAK.     

Dietary isothiocyanate iberin inhibits growth and induces apoptosis in human glioblastoma cells

In this study, we evaluated the antiproliferative and proapoptotic effects of the isothiocyanate iberin, a bioactive agent in Brassicaceae species, in human glioblastoma cells. The human glioblastoma cell cultures were treated with different concentrations of iberin and tested for growth inhibition, cytotoxicity, induction of apoptosis, and activation of caspases. Iberin inhibited growth of tumor cells in cell proliferation assays, enhanced cytotoxicity, and induced apoptosis by activation of caspase-3 and caspase-9. Findings from this study could provide a basis for potential usefulness of the diet-derived isothiocyanate iberin as a promising therapeutic micronutrient in the prevention/intervention of brain tumors.     

核糖体蛋白S3a在肿瘤细胞增殖分化和凋亡调控作用的研究概况

目的综述核糖体蛋白S3a在肿瘤细胞增殖分化和凋亡调控作用的研究进展。方法参阅近几年国内外相关文献,对核糖体蛋白S3a的结构、功能及其异常表达对肿瘤细胞增殖分化和凋亡的调控等方面的进展进行归纳总结。结果与结论核糖体蛋白S3a除了在蛋白质合成中起重要作用外,还有独特的核糖体外功能。其在多种肿瘤细胞中高表达,通过调控癌基因和抑癌基因的表达可影响肿瘤细胞的增殖分化与凋亡。     

miR-200b 靶向DNMT3A 抑制非小细胞肺癌细胞增殖与诱导凋亡

摘要： 目的 探讨miR-200b是否通过靶向调控DNMT3A抑制人非小细胞肺癌A549细胞增殖与诱导凋亡。方法 运用qRT-PCR检测miR-200b在不同非小细胞肺癌细胞株中的表达; 将miR-200b mimics、 scramble、 DNMT3A-siRNA、 control-siRNA分别转染于A549细胞, 其中scramble与control-siRNA分别作为miR-200b mimics与DNMT3A-siRNA 的阴性对照组。采用 Western blot 检测 A549 细胞中 DNMT3A 蛋白表达; 采用 MTT 与 AnnexinV-FITC/PI 染色法分别检测 A549 细胞的增殖与凋亡, 比较 miR-200b mimics 与 DNMT3A-siRNA 对 A549 细胞增殖与凋亡的影响。结果 qRT-PCR 结果显示, miR-200b 在非小细胞肺癌 A549、 H1299、 L78、 H460 细胞中的表达均明显低于正常人支气管上皮 16HBE 细胞, 其中以 A549 细胞下调最为明显 （P < 0.05）。Western blot 结果显示, 外源过表达 miR-200b 或沉默 DNMT3A 能明显下调 A549 细胞中 DNMT3A 蛋白的水平。MTT 结果显示, 转染 miR-200b mimics 或沉默 DN⁃ MT3A 48 h、 72 h、 96 h 后, 反映细胞增殖的光密度 （OD） 值与各自阴性对照组比较明显减小 （P < 0.05）。AnnexinV- FITC/PI 染色结果显示, 转染 miR-200b mimics 或沉默 DNMT3A 后, A549 细胞的凋亡率分别为 （23.33%±0.90%、 20.41%±0.70%） 均高于各自阴性对照组 （5.28%±0.55%、 5.68%±0.47%, P < 0.05）。结论 miR-200b通过下调DNMT3A 抑制人非小细胞肺癌细胞增殖与诱导凋亡。     

S-allylcysteine,a garlic derivative,suppresses proliferation and induces apoptosis in human ovarian cancer cells in vitro

Aim： To investigate the effects of S-allylcysteine （SAC）, a water-soluble garlic derivative, on human ovarian cancer cells in vitro. Methods： Human epithelial ovarian cancer cell line A2780 was tested. Cell proliferation was examined with CCK-8 and colony formation assays. Cell cycle was analyzed with flow cytometry. Cell apoptosis was studied using Hoeohst 33258 staining and Annexin V/PI staining with flow cytometry. The migration and invasion of A2780 cells were examined with transwell and wound healing assays. The expression of relevant proteins was detected with Western blot assays. Results： SAC （1-100 mmol/L） inhibited the proliferation of A2780 cells in dose- and time-dependent manners （the ICsovalue was approximately 25 mmoVL at 48 h, and less than 6.25 mmol/L at 96 h）. Furthermore, SAC dose-dependently inhibited the colony formation of A2780 cells. Treatment of A2780 cells with SAC resulted in G/S phase arrest and induced apoptosis, accompanied by decreased expression of pro-caspase-3, Parp-1 and Bcl-2, and increased expression of active caspase-3 and Bax. SAC treatment significantly reduced the migration of A2780 cells, and markedly decreased the protein expression of Wnt5a, p-AKT and c-Jun, which were the key proteins involved in proliferation and metastasis. Conclusion： SAC suppresses proliferation and induces apoptosis in A2780 ovarian cancer cells in vitro.     

吡格列酮抑制人胶质瘤SNB19细胞生长及诱导其凋亡

目的 探讨吡格列酮(PGZ)对人神经胶质瘤细胞株SNB19增殖和凋亡的影响及其机制。方法 用MTT、流式细胞仪、Tunel和Western blot分析技术。结果 吡格列酮能够抑制SNB19细胞增殖和诱导其凋亡,同时伴有Bax表达的增高和pRb、Bcl-2表达的降低。结论 PGZ能够在体外抑制胶质瘤SNB19细胞的生长并诱导其凋亡,提示PGZ有可能成为一种新的治疗胶质瘤药物。     

Ropivacaine inhibits proliferation,invasion, migration and promotes apoptosis of papillary thyroid cancer cells via regulating ITGA2 expression

The present study aimed to investigate the roles of Ropivacaine in papillary thyroid cancer (PTC) and identify the possible mechanisms. The expression of integrin alpha-2 (ITGA2) in TC cell lines was tested using Western blotting and RT-qPCR. Subsequently, the level of ITGA2 in human PTC cell line (TPC-1) was measured following intervention with a series of concentrations of Ropivacaine. Then, cell counting kit-8 (CCK-8) assay and colony formation assay were executed for detecting proliferation of cells after transfection with ITGA2 pcDNA3.1. The expression of proliferation-related protein was determined by Western blotting. Additionally, the abilities of TPC-1 cell invasion and migration were examined using Transwell assay and scratch wound healing assay. Apoptosis of TPC-1 cells was analyzed using TUNEL assay and the expressions of apoptosis-related proteins were tested via West blotting. The results suggested that ITGA2 was highly expressed in TC cell lines, especially in TPC-1 cells. Ropivacaine decreased the expression of ITGA2 in a dose-dependent manner. Moreover, after treatment with Ropivacaine, cell proliferation was inhibited accompanied by changes of proliferation-related protein expressions, which was reversed following co-transfection with ITGA2 pcDNA3.1. Furthermore, Ropivacaine concentration-dependently suppressed invasion and migration of TPC-1 cells, whereas these inhibitory effects were attenuated after ITGA2 overexpression. Furthermore, apoptosis was promoted, coupled with a decrease of Bcl-2 expression and increases of Bax, cleaved caspase-3 and cleaved caspase-9 expression, in Ropivacaine-treated TPC-1 cells, which was restored following ITGA2 overexpression. These findings demonstrated that Ropivacaine could suppress proliferation, invasion, migration, and accelerate apoptosis of PTC cells via regulating ITGA2 expression.     

MAPK/ERK通过诱导纤维蛋白A磷酸化抑制垂体瘤细胞增殖、凋亡、迁移及相关信号转导

目的 探讨MAPK/ERK对垂体瘤细胞增殖、凋亡、迁移及相关信号转导的影响.方法 通过CCK-8法确定SB203580(MAPK/ERK抑制剂)的最佳安全浓度,然后将体外培养的垂体瘤细胞分为SB203580组和对照组,通过CCK-8法检测垂体瘤细胞的增殖情况,流式细胞术检测垂体瘤细胞的细胞凋亡百分数,RT-PCR和We...     

X H-211对3T3-L1细胞增殖、分化和凋亡的影响

目的：研究异黄酮脂肪酸酯类化合物 XH‐211对前脂肪细胞株3T3‐L1增殖、分化和凋亡的影响。方法体外培养3T3‐L1细胞,加入不同浓度XH‐2111×10‐4、1×10‐5、1×10‐6和1×10‐7 mol／L ;同时另设DMSO空白对照组。采用MTT法和流式细胞术分别检测3T3‐L1细胞增殖和凋亡情况,油红染色法观察3T3‐L1细胞分化过程产脂量的变化。结果与空白对照组相比,XH‐211呈浓度依赖性地抑制3T3‐L1细胞增殖和油脂的产生,诱导3T3‐L1细胞和分化成熟的3T3‐L1细胞凋亡（P＜0．05）。结论 XH‐211通过诱导3T3‐L1细胞凋亡,抑制细胞增殖,减少分化过程中的产脂量,从而减少脂肪堆积,降低血脂。     

Synergistic suppression of noscapine and conventional chemotherapeutics on human glioblastoma cell growth

Aim:

Noscapine (NOS) is a non-narcotic opium alkaloid with anti-tumor activity. The aim of this study was to investigate the effects of the combination of NOS with conventional chemotherapeutics temozolamide (TMZ), bis-chloroethylnitrosourea (BCNU), or cisplatin (CIS)on human glioblastoma cells.

Methods:

U87MG human glioblastoma cells were examined. Cell proliferation was quantified using MTT assay. Western blotting and flow cytometry were used to examine apoptosis and the expression of active caspase-3 and cleaved PARP. Mouse tumor xenograft model bearing U87MG cells was treated with TMZ (2 mg·kg⁻¹·d⁻¹, ip) or CIS (2 mg/kg, ip 3 times a week) alone or in combination with NOS (200 mg·kg⁻¹·d⁻¹, ig) for 3 weeks. Immunohistochemistry was used to investigate the expression of active caspase-3 and Ki67 following treatment in vivo. The safety of the combined treatments was evaluated based on the body weight and histological studies of the animal''s organs.

Results:

NOS (10 or 20 mol/L) markedly increased the anti-proliferation effects of TMZ, BCNU, and CIS on U87MG cells in vitro. The calculated combination index (CI) values of NOS-CIS, NOS-TMZ, and NOS-BCNU (20 μmol/L) were 0.45, 0.51, and 0.57, respectively, demonstrating synergistic inhibition of the drug combinations. In tumor xenograft models, combined treatment with NOS robustly augmented the anti-cancer actions of TMZ and CIS, and showed no detectable toxicity. The combined treatments significantly enhanced the apoptosis, the activated caspase-3 and PARP levels in U87MG cells in vitro, and reduced Ki67 staining and increased the activated caspase-3 level in the shrinking xenografts in vivo.

Conclusion:

NOS synergistically potentiated the efficacy of FDA-approved anti-cancer drugs against human glioblastoma cells, thereby allowing them to be used at lower doses and hence minimizing their toxic side effects.     

慢病毒介导的PRL-3 shRNA对结肠癌细胞增殖、侵袭、凋亡的影响

目的 观察慢病毒介导的 shRNA 沉默肝再生磷酸酶-3（PRL-3）基因对结肠癌 SW480 细胞增殖、侵袭、凋亡的影响。方法 实验分组为空白对照组、阴性对照组、转染组。将携带 PRL-3 shRNA 的慢病毒载体转染结肠癌SW480 细胞,建立稳定沉默 PRL-3 的细胞株,real-time PCR 检测转染后 PRL-3 mRNA 的相对表达水平。采用 MTT法、平板克隆形成实验检测转染后细胞增殖能力;采用 Transwell 侵袭实验、侵袭小室法检测转染后细胞迁移及侵袭能力;采用流式细胞术检测转染后细胞凋亡率变化。结果 稳定沉默 PRL-3 的细胞株构建成功,转染组 PRL-3mRNA 的相对表达水平低于空白对照组、阴性对照组（P＜0.05）,空白对照组、阴性对照组比较差异无统计学意义。PRL-3 shRNA 转染 SW480 细胞 72 h 后,转染组与空白对照组、阴性对照组比较,细胞增殖能力受到抑制,转染 120 h时最明显（P＜0.05）。转染组克隆形成能力较空白对照组、阴性对照组下降（P＜0.05）。转染组与空白对照组、阴性对照组比较,细胞迁移、侵袭能力下降,凋亡率增加（P＜0.05）。结论 结肠癌 SW480 细胞转染 PRL-3 shRNA 可减少 PRL-3 的表达,有效抑制 SW480 细胞增殖,促进其凋亡,PRL-3 可能成为治疗结肠癌的靶基因。     

紫杉醇对绒癌JEG-3细胞增殖与凋亡的影响

目的该文围绕紫杉醇对人绒毛膜癌细胞JEG-3的增殖与凋亡的影响进行研究。方法通过MTT法检测细胞增殖及流式细胞术检测不同浓度紫杉醇作用下细胞的凋亡及其对细胞周期的影响。结果随紫杉醇浓度的增加或时间的延长,JEG-3细胞增殖抑制率及凋亡率均增大,各组间比较有统计学意义（P〈0.05）。结论该次实验的结果显示紫杉醇在抑制绒癌细胞JEG-3增殖及促凋亡方面均有显著的作用。     

前列腺源性ETS因子对HCT116细胞增殖与凋亡的影响

目的 探讨前列腺源性ETS因子(prostate-derived Ets factor,PDEF)对HCT116细胞增殖与凋亡的影响.方法 体外培养HCT116细胞,分成空白对照组,空质粒组和重组表达质粒组,空质粒组转染不含PDEF的空载体,重组表达质粒组转染PDEF重组表达质粒.荧光显微镜下观察PDEF重组质粒表达情况;RT-PCR、Western blot检测3组细胞PDEF mRNA和蛋白的表达情况;CCK8法检测细胞增殖情况;流式细胞仪检测细胞凋亡率.结果 荧光显微镜下空质粒组和重组表达质粒组HCT116细胞均可观察到绿色荧光蛋白的表达,表明质粒已成功转染HCT116细胞;RT-PCR结果显示,与空质粒组相比,重组质粒组PDEF基因表达增高263倍;Western blot结果可见重组质粒组有PDEF蛋白的表达,而空质粒组和对照组没有;CCK8法检测发现PDEF基因的表达能够明显抑制HCT116细胞的增殖(P＜0.05);流式细胞仪结果显示重组质粒组细胞凋亡率显著高于对照组和空质粒组(P＜0.05).结论 在HCT116细胞中表达前列腺上皮源性ETS转录因子,可以抑制细胞的增殖和促进细胞凋亡.     

吉非替尼通过调控hPTTG1表达抑制卵巢癌细胞增殖并诱导凋亡的机制研究

目的：吉非替尼和人垂体瘤转化基因1（hPTTG1）对卵巢癌细胞增殖和凋亡的影响及作用机制。方法：以不同浓度吉非替尼干预A2780细胞,MTT法检测细胞增殖。流式细胞仪、qRT-PCR和Western blot实验检测20 μmol·L^-1吉非替尼对A2780细胞凋亡及细胞中hPTTG1表达的影响。敲减hPTTG1或过表达hPTTG1联合20 μmol·L^-1吉非替尼处理,qRT-PCR和Western blot、MTT和流式细胞术检测细胞中hPTTG1表达及细胞增殖、凋亡情况变化。结果：吉非替尼能够有效抑制A2780细胞增殖（P<0.05）,呈浓度依赖性。以20 μmol·L^-1吉非替尼干预A2780细胞,细胞凋亡率升高（P<0.05）,细胞中hPTTG1在mRNA和蛋白水平的表达显著下调（P<0.05）。将hPTTG1 siRNA转染至A2780细胞后,细胞hPTTG1表达水平降低（P<0.05）,细胞增殖抑制率和凋亡率增大（P<0.05）;过表达hPTTG1可减弱吉非替尼对A2780细胞增殖的抑制及凋亡的促进作用。结论：吉非替尼可抑制A2780细胞增殖并诱导凋亡,这一结果可能是通过抑制hPTTG1表达来完成。     

AICAR inhibits proliferation and induced S-phase arrest, and promotes apoptosis in CaSki cells

Aim： The aim of the present study was to determine the effect of 5-aminoimidazole- 4-carboxamide-ribonucleoside （A/CAR） on proliferation, cell cycle, and apoptosis in the human epithelial cervical cancer cell line CaSki cells. Methods： Cell count and 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay were used to determine cell proliferation and viability. Hoechst 33258 staining was con- ducted to distinguish the apoptotic cells. Cell cycle and Annexin-V/propidium iodide staining were analyzed by fluorescence-activated cell sorting （FACS）. A Western blot assay was used to evaluate the expression of AKT （also known as protein kinase B）, mammalian target of rapamycin （mTOR）, p53, and extracellular signal-regulated kinase （ERK）. Results： A/CAR （500 pmol/L） significantly inhibi- ted the proliferation of CaSki cells treated for 24, 48, and 72 h as determined by cell count. The cells at the Gl and G2 phases were dramatically decreased while cells at the S phase were increased in response to A/CAR treatment for 24, 48, and 72 h, The MTF assay showed less viable cells and Hoechst fluorescent staining showed more apoptotic cells upon AICAR stimulation. The results of the Annexin-V staining demonstrated a time-dependent increase of apoptosis in cells treated with A/CAR for 24, 36, and 48 h. Furthermore, AICAR activated caspase-3 in a time-dependent manner. It was also found that AICAR inhibited the phosphory- lation of AKT and mTOR, which are important kinases regulating cell growth and survival. AICAR stimulation obviously increased the expression of the tumor suppressor p53 and the phosphorylation of ERK. Conclusion： A/CAR inhibited proliferation and induced S phase arrest and promoted apoptosis in CaSki cells, which might be mediated by the dowrtregulation of the AKT/mTOR pathway and the upregulation of the p53/ERK pathway.     

天花粉蛋白对人宫颈癌HeLa细胞增殖和细胞调亡的影响

天花粉蛋白(Trichosanthin,TCS)是从葫芦科植物栝(蒌)的块根中提取出来的一种单链核糖体失活蛋白(ribosome-in-activating protein,RIP),主要引发中期流产,抑制爱滋病病毒(HIV)增殖,调节人体免疫功能等多方面的作用[1].     

尿石素C对胶质母细胞瘤生物学行为的调控作用及机制

目的　探讨尿石素C（UC）对胶质母细胞瘤（GBM）U251和U87 MG细胞生物学行为的影响及调控机制。方法　将U251和U87 MG细胞分为0、20、40、80、120和160 μmol/L UC处理组,通过CCK-8法分别检测各组细胞24、48和72 h时增殖率。将细胞分为0、40、80和120 μmol/L UC处理组,通过细胞克隆形成实验检测细胞克隆形成能力,划痕愈合实验检测24 h和48 h时细胞迁移能力,Transwell小室侵袭实验检测24 h时细胞侵袭能力,流式细胞术检测24 h时细胞凋亡率及细胞周期,Western blot检测AMP依赖的蛋白激酶（AMPK）、细胞外调节蛋白激酶（ERK）和丝裂原活化蛋白激酶p38（p38 MAPK）蛋白表达水平和磷酸化水平。结果　与0 μmol/L组比较,U251细胞中不同浓度UC组于48 h开始增殖率均降低,而U87 MG细胞中40 μmol/L及以上浓度组于48 h时增殖率开始降低（P＜0.05）。与0 μmol/L组比较,40、80和120 μmol/L组U251和U87 MG细胞的克隆形成率、24 h和48 h时的细胞迁移能力和24 h时的细胞侵袭能力降低,且均呈浓度依赖性（P＜0.05）。与0 μmol/L组比较,120 μmol/L组U251和U87 MG细胞凋亡率均增加,40、80和120 μmol/L组U251和U87 MG细胞周期均阻滞在S期（P＜0.05）。与0 μmol/L组比较,40、80和120 μmol/L组U251和U87 MG细胞中p-AMPK和p-p38 MAPK水平均升高,80和120 μmol/L组U251细胞中p-ERK水平升高,但仅120 μmol/L组U87 MG细胞p-ERK水平升高（P＜0.05）。结论　一定浓度的UC可抑制GBM细胞增殖、迁移和侵袭,诱导肿瘤细胞凋亡和细胞周期阻滞,其机制可能与调控AMPK/ERK/p38 MAPK信号通路有关。